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Clinacanthus nutans is a traditional medicinal herb that is applied for the therapy of snake bites, skin infection, herpes infection, burns, scalds, dysentery, and diabetes. Clinacanthus nutans is also used to treat several cancers, including breast, cervical, colon, gastric, head and neck, liver, lung, pancreatic, and skin cancers, as well as lymphoma and leukemia; however, the underlying mechanisms of its anticancer activity remained undetermined. We searched PubMed and Google with key words “Clinacanthus nutans and cancer” and collected recent papers of Clinacanthus nutans with anticancer activity. We focused on the preparation, effects, and action mechanisms of Clinacanthus nutans extracts on various types of cancers. We hope that this mini review can help update our knowledge about active components, effects, and molecular mechanisms of extracts from this promising herb Clinacanthus nutans for ongoing studies and speed up its clinical application in the future.

Silver nanoparticles (AgNPs) previously synthesised using leaf (AgNP-L) and stem (AgNP-S) extracts of Clinacanthus nutans (C. nutans) were tested to evaluate antimicrobial, antioxidant, and cytotoxicity activities. The AgNPs showed good inhibition against bacteria, but not fungi. The inhibition results showed the highest activity against Staphylococcus aureus (S. aureus) with 11.35 mm (AgNP-L) and 11.52 mm (AgNP-S), while the lowest inhibition was against Escherichia coli (E. coli) with 9.22 mm (AgNP-L) and 9.25 mm (AgNP-S) in the disc diffusion method. The same trend of results was noted in the well diffusion method. The IC50 of AgNP-L and AgNP-S in 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays was 417.05 μg/mL and 434.60 μg/mL, as well as 304.31 μg/mL and 326.83 μg/mL, respectively. Ferric reducing power (FRAP) assay showed that AgNP-L [872.389 μmol/L Fe(II)] and AgNP-S [612.770 μmol/L Fe(II)] exhibited significantly ( p p

Changes in kidney homeostasis due to diabetes can cause oxidative stress which then caused tissue damage that leads to diabetic nephropathy. Clinacanthus nutans extract is known to contain antioxidants that are reported to play an important role in the body’s defense system against oxidative stress to minimize tissue damage. This study aims to know the effect of Clinacanthus nutans leaf extract administration on kidney histopathological features of the diabetic rats model. A total of 35 rats were induced by streptozotocin which then divided into 5 groups and given Clinacanthus nutans extract with a dose of 75 mg/kgBW, 150 mg/kgBW and 300 mg/kgBW then compare with CMC-Na as control and metformin as standard drug for 14 days. The kidney histopathology was evaluated under a light microscope against the damage criteria that occurred in the proximal tubules of the kidney. As the result the least amout of kidney damage was on treatment group at dose 300 mg/kgBW, followed by extract with dose 75 mg/kgBW, 150 mg/kgBW, and metformin. In conclusion Clinacanthus nutans extract with a dose of 75 mg/kgBW, 150 mg/kgBW, and 300 mg/kgBW can improve the kidney histopathological feature of the diabetic rats model. Keywords: diabetes mellitus; histopathology; diabetic nephropathy; Clinacanthus nutans

Context Atherosclerosis predisposes individuals to adverse cardiovascular events. Clinacanthus nutans L. (Acanthaceae) is a traditional remedy used for diabetes and inflammatory conditions. Objectives To investigate the anti-atherosclerotic activity of a C. nutans leaf methanol extract (CNME) in a type 2 diabetic (T2D) rat model induced by a high-fat diet (HFD) and low-dose streptozotocin. Materials and methods Sixty male Sprague-Dawley rats were divided into five groups: non-diabetic fed a standard diet (C), C + CNME (500 mg/kg, orally), diabetic fed an HFD (DM), DM + CNME (500 mg/kg), and DM + Metformin (DM + Met; 300 mg/kg). Treatment with oral CNME and metformin was administered for 4 weeks. Fasting blood glucose (FBG), serum lipid profile, atherogenic index (AI), aortic tissue superoxide dismutase levels (SOD), malondialdehyde (MDA), and tumour necrosis factor-alpha (TNF-α) were measured. The rats’ aortas were stained for histological analysis and intima-media thickness (IMT), a marker of subclinical atherosclerosis. Results The CNME-treated diabetic rats had reduced serum total cholesterol (43.74%; p = 0.0031), triglycerides (80.91%; p = 0.0003), low-density lipoprotein cholesterol (56.64%; p = 0.0008), AI (51.32%; p

Introduction: Cervical cancer is a leading cause of death in women. Current cancer treatment comes with side effects. Clinacanthus nutans has been known traditionally to treat cancer. This study was aimed to characterize C. nutans standardized fraction (SF1) and to investigate its anticancer mechanism against SiHa cells. Materials and Methods: SF1 was produced by optimized methodology for bioassay-guided fractionation. Fourier transform infrared (FTIR) spectroscopy and liquid chromatography–mass spectrometry (LC-MS) were carried out to characterize the SF1. SF1 was screened for cytotoxicity activity toward HeLa, SiHa, and normal cells (NIH) cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. The anticancer mechanism of SF1 was evaluated toward SiHa cells, which showed highest cytotoxicity toward SF1 treatment. The mechanism includes cell cycle progression and protein expression, which was detected using specific antibody-conjugated fluorescent dye, p53-FITC, by flow cytometry. Results: Major constituents of SF1 were alkaloids with amines as functional group. SF1 showed highest cytotoxic activity against SiHa (half-maximal inhibitory concentration [IC50] < 10 µg/mL) compared to HeLa cells. Cytoselectivity of SF1 was observed with no IC50 detected on normal NIH cells. On flow cytometry analysis, SF1 was able to induce apoptosis on SiHa cells by arresting cell cycle at G1/S and upregulation of p53 protein. Conclusion: SF1 showed anticancer activity by inducing apoptosis through arrested G1/S cell cycle checkpoint–mediated mitochondrial pathway.

Neuroinflammation has been shown to exacerbate ischemic brain injury, and is considered as a prime target for the development of stroke therapies. Clinacanthus nutans Lindau (C. nutans) is widely used in traditional medicine for treating insect bites, viral infection and cancer, due largely to its anti-oxidative and anti-inflammatory properties. Recently, we reported that an ethanol extract from the leaf of C. nutans could protect the brain against ischemia-triggered neuronal death and infarction. In order to further understand the molecular mechanism(s) for its beneficial effects, two experimental paradigms, namely, in vitro primary cortical neurons subjected to oxygen-glucose deprivation (OGD) and in vivo rat middle cerebral artery (MCA) occlusion, were used to dissect the anti-inflammatory effects of C. nutans extract. Using promoter assays, immunofluorescence staining, and loss-of-function (siRNA) approaches, we demonstrated that transient OGD led to marked induction of IL-1β, IL-6 and TNFα, while pretreatment with C. nutans suppressed production of inflammatory cytokines in primary neurons. C. nutans inhibited IL-1β transcription via preventing NF-κB/p65 nuclear translocation, and siRNA knockdown of either p65 or IL-1β mitigated OGD-mediated neuronal death. Correspondingly, post-ischemic treatment of C. nutans attenuated IκBα degradation and decreased IL-1β, IL-6 and TNFα production in the ischemic brain. Furthermore, IL-1β siRNA post-ischemic treatment reduced cerebral infarct, thus mimicking the beneficial effects of C. nutans. In summary, our findings demonstrated the ability for C. nutans to suppress NF-κB nuclear translocation and inhibit IL-1β transcription in ischemic models. Results further suggest the possibility for using C. nutans to prevent and treat stroke patients.

Objective: The present study aims to evaluate the antioxidant, antimicrobial and cytotoxic properties of the secondary metabolites and proteins extracted from Clinacanthus nutans. Methods: Methanol (ME) and acetone (AC) and also protein (PE) extracts were obtained from Clinacanthus nutans. Antioxidant activity of sample extracts was examined by using 1,1-Diphenyl-2-Picrydyl Hydrazyl (DPPH) assay and antimicrobial activity was examined by using the agar diffusion method against Bacillus subtilis, Bacillus cereus, Streptococcus pyrogenes, Escherichia coli, and Pseudomonas aeroginosa bacterial strain. The cytotoxic potential was determined by brine shrimp lethality assay. Results: Methanol extract (ME) demonstrated to have the highest antioxidant activity with IC50 value of 98.84 μg/ml, followed by acetone extract (AE) with IC50 value of 134.83µg/ml, and protein extract (PE) with IC50 value of 353.49 µg/ml. Among all of the sample extracts, only methanol crude extracts (ME) displayed moderate inhibition against Gram-positive B. cereus (7.33±1.15 mm) and S. pyrogenes (8.67±0.57 mm) at concentration of 100 mg/ml, while both acetone (AE) and protein (PE) extracts had no activity against all tested microorganisms. All extracts from methanol (ME), acetone (AE) and protein (PE) exhibited significant cytotoxic activity against brine shrimps at LC50 7.2 μg/ml, 1.42 μg/ml and 70.6 μg/ml respectively. Conclusion: These data proved that methanol extract from the leaves of Clinacanthus nutans is the most potent among all samples tested and has the potential to be developed as an antioxidant, antimicrobial and cytotoxic agents. Whilst, acetone and protein crude extracts have potent antioxidant and cytotoxicity activities.

Diabetes mellitus is associated with endothelial dysfunction; it causes progressive vascular damage resulting from an impaired endothelium-dependent vasorelaxation. In the diabetes state, presence of hyperglycemia and insulin resistance predisposes to endothelial dysfunction. Clinacanthus nutans, widely used as a traditional medicine for diabetes is reported to have hypoglycemic, hypolipidemic, antioxidant, and anti-inflammatory properties. However, the possibility of C. nutans affecting the vascular endothelial function in diabetes remains unclear. This study was aimed at evaluating the effects of C. nutans methanolic leaves extract (CNME) on endothelial function in a type 2 diabetes (T2DM) rat model. Sixty male Sprague-Dawley rats were divided into five groups (n=12 per group): nondiabetic control, nondiabetic treated with four weeks of CNME (500 mg/kg/daily), untreated diabetic rats, diabetic treated with metformin (300 mg/kg/daily), and diabetic treated with CNME (500 mg/kg/daily). T2DM was induced by a single intraperitoneal injection of low-dose streptozotocin (STZ) to rats fed with high-fat diet (HFD). Endothelial-dependent and endothelial-independent relaxations and contractions of the thoracic aorta were determined using the organ bath. Aortic endothelial nitric oxide synthase (eNOS) expression was determined using Western blotting. Endothelial-dependent relaxation was reduced in diabetic rats. Both diabetic groups treated with CNME or metformin significantly improved the impairment in endothelium-dependent vasorelaxation; this was associated with increased expression of aortic eNOS protein. CNME- and metformin-treated groups also reduced aortic endothelium-dependent and aortic endothelium-independent contractions in diabetics. Both of these diabetic-treated groups also reduced blood glucose levels and increased body weight compared to the untreated diabetic group. In conclusion, C. nutans improves endothelial-dependent vasodilatation and reduces endothelial-dependent contraction, thus ameliorating endothelial dysfunction in diabetic rats. This may occur due to its effect on increasing eNOS protein expression.

Clinacanthus nutans (Acanthaceae), locally known as Sabah snake grass, is popularly taken as prevention as well as treatment for cancer in Malaysia, despite lack of concrete clinical evidence. However, it is crucial to evaluate potential antagonistic, additive, or synergistic interactions that may result from the co-treatment of this plant in chemotherapy. In this study, we demonstrate the drug-herb interaction using combination treatment of C. nutans extracts and cyclophosphamide on the WRL 68 cell line. Materials and Methods: Aqueous and 70% ethanol extract of C. nutans leaves were prepared using decoction and maceration methods. MTT assay was used to test single treatment as well as the combination of C. nutans extract and cyclophosphamide. Phytochemical profiling and flavonoid were identified using HPLC. Results: C. nutans ethanolic extract exhibits low antiproliferative activity. A combination of ethanolic extract of C. nutans and cyclophosphamide at various concentrations resulted in antagonism with combination index values of 1.413, 1.482, and 1.525. Flavonoids identified in the phytochemical profile of both extracts were schaftoside, isoorientin, orientin, and vitexin. High flavonoids level in C. nutans ethanolic extract could potentially interact with metabolic enzymes of the cell line, which might have affected metabolism and activated the cyclophosphamide. The in vitro data suggested that there was a potential for drug-herb interactions, which could negatively affect the chemotherapeutic outcomes. Further investigation should be done in vivo and clinical research model.

Objective: This study aimed to evaluate acute and sub-acute oral toxicity of ethanol extract of Clinacanthus nutans leaves in Swiss mice. Methods: Acute oral toxicity study was performed as per OECD-423 guidelines. Sub-acute oral toxicity study was performed as per OECD-407 guidelines. The extract was dissolved in 10% dimethyl sulfoxide and administered orally, while the control group received only the vehicle. Results: The acute oral toxicity test on mice showed that this extract was well tolerated up to LD50 5000 mg/kg body weight/day oral dosage level and non-toxic to mice under the present experimental conditions. The sub-acute toxicity study was carried out on mice with the oral dosage of the extract from 100 mg/kg–500 mg/kg body weight/day and 5000 mg/kg body weight/day for 28 d. The results showed that this extract did not induce death or adverse effects in activity, feed consumption or body weight gain. There were not significant changes in heamotological and biochemical parameters between control and experiment groups. Conclusion: Thus, Clinacanthus nutans leaf has a very low toxicity value.

PurposeThe purpose of this paper is to evaluate the antioxidant activity and toxicity of Clinacanthus nutans leaves from three conventional extraction methods, i.e. maceration, Soxhlet and magnetic stirring.Design/methodology/approachTotal flavonoid content (TFC) and phenolic content (TPC) were determined using colorimetric method of aluminum chloride and Folin-Ciocalteu (FC) assay, respectively. Antioxidant property of C. nutans was evaluated using 2,2'-diphenyl-1-pierylhydrazyl (DPPH) free radical scavenging assay. Cytotoxic activity of C. nutans against brine shrimp was evaluated based on LC50 (lethality concentration) after 24 h exposure to the plant extract.FindingsThe highest TPC of C. nutans was observed with Soxhlet extraction method (98.87 ± 10.43 mg of gallic acid equivalents (GAE/g) followed by maceration (68.77 ± 2.45 mg of GAE/g) and magnetic stirring (46.75 ± 2.45 mg of GAE/g). Interestingly, remarkable highest TFC was observed with magnetic stirring (568.90 ± 4.85 mg of rutin equivalent (RE)/g) followed by maceration (249.60 ± 2.79 mg of RE/g) and Soxhlet (174.8 ± 1.74 mg of RE/g). On the other hands, the extract obtained using maceration method showed the highest antioxidant activity (IC50: 14.18 mg/mL compared to ascorbic acid 144.36 µg/mL). Cytotoxicity of C. nutans from all extraction methods showed similar LC50 values with maceration (3.81 mg/mL), Soxhlet (2.61 mg/mL) and magnetic stirring (4.56 mg/mL), respectively.Originality/valueBoth phenolic and flavonoids are responsible for the antioxidant activity, of C. nutans extracts. Based on Meyer's toxicity index, all extracts were nontoxic (LC50>1 mg/mL).

Clinacanthus nutans is a great medicinal herb with high polyphenols content and had been used in its infusion form to treat illness. In this paper, microwave-assisted extraction of C. nutans tea infusion was investigated by using central composite design combined with response surface methodology. The influence factors of microwave power (80 - 800 W); brewing time duration (0 – 240 s) and extraction time duration (60 s – 300 s) on total phenolic content (TPC), total flavonoid content (TFC), antioxidant activity (DPPH) and acetylcholinesterase inhibition activity (AChE) were investigated. The optimum extraction conditions were determined as follows: microwave power X1, 560 W; brewing duration, X2, 60 s and extraction time, X3, 240 s. Under this optimal extraction conditions, the responses were as followed: Total phenolic content (251.58±10.98 mg GAE/L); Total flavonoid content (102.21±5.16 mg RE/L); DPPH (62.22±0.03%) and acetylcholinesterase inhibition activity (AChE) of 24.77±0.12%. Results demonstrated that MAE could be used to extract bioactive constituents having antioxidant and anti-acetylcholinesterase properties that may benefit in Alzheimer’s disease.

BACKGROUND: Osteoporotic-osteoarthritis is an incapacitating musculoskeletal illness of the aged. OBJECTIVES: The anti-inflammatory and anti-catabolic actions of Diclofenac were compared with apigenin-C-glycosides rich Clinacanthus nutans (CN) leaf extract in osteoporotic-osteoarthritis rats. METHODS: Female Sprague Dawley rats were randomized into five groups (n = 6). Four groups were bilateral ovariectomised for osteoporosis development, and osteoarthritis were induced by intra-articular injection of monosodium iodoacetate (MIA) into the right knee joints. The Sham group was sham-operated, received saline injection and deionized drinking water. The treatment groups were orally given 200 or 400 mg extract/kg body weight or 5 mg diclofenac /kg body weight daily for 28 days. Articular cartilage and bone changes were monitored by gross and histological structures, micro-CT analysis, serum protein biomarkers, and mRNA expressions for inflammation and catabolic protease genes. RESULTS: HPLC analysis confirmed that apigenin-C-glycosides (shaftoside, vitexin, and isovitexin) were the major compounds in the extract. The extract significantly and dose-dependently reduced cartilage erosion, bone loss, cartilage catabolic changes, serum osteoporotic-osteoarthritis biomarkers (procollagen-type-II-N-terminal-propeptide PIINP; procollagen-type-I-N-terminal-propeptide PINP; osteocalcin), inflammation (IL-1β) and mRNA expressions for nuclear-factor-kappa-beta NF-κβ, interleukin-1-beta IL-1β, cyclooxygenase-2; and matrix-metalloproteinase-13 MMP13 activities, in osteoporotic-osteoarthritis rats comparable to Diclofenac. CONCLUSION: This study demonstrates that apigenin-C-glycosides at 400 mg CN extract/kg (about 0.2 mg apigenin-equivalent/kg) is comparable to diclofenac in suppressing inflammation and catabolic proteases for osteoporotic-osteoarthritis prevention. [Figure: see text] ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s40199-020-00343-y) contains supplementary material, which is available to authorized users.

Limitations in existing anticoagulants have prompted a search for the natural origin of anticoagulants. Since there is an increasing need to source for pharmacological and medicinal materials from a plant source, thus an exploratory effort towards identifying and characterizing new anticoagulants from plants is worthwhile. Clinacanthus nutans (C. nutans) commonly known as Sabah snake grass belongs to Acanthaceae family was chosen since it has therapeutic and medicinal benefits. This plant possessed anti-oxidant, anti-microbial, anti-inflammatory, and anti-diabetic activities. This study was carried out to identify the anticoagulant activities of C. nutans leaves in aqueous and methanol extracts at different concentrations based on prothrombin time (PT) and activated partial thromboplastin time (aPTT). The dried leaves of C. nutans were ground into a fine powder and extracted using aqueous and methanol. Anticoagulant assays of PT and aPTT were done on three different concentrations of C. nutans aqueous and methanol extracts (10 mg/mL, 20 mg/mL, and 30 mg/mL) and control group respectively. PT and aPTT of plasma samples were prolonged in both types of extracts (p

Clinacanthus nutans Lindua (C. nutans), a strong antiviral traditional medicine, can be used to treat condyloma acuminata (CA) caused by the human papillomavirus (HPV). However, its molecular mechanism for CA elimination is unknown. Herein, we conducted a randomized clinical trial to evaluate the effectiveness of C. nutans and its molecular mechanism compared with podophyllin, the gold standard treatment. Using a randomized block design, six patients were treated with C. nutans and podophyllin for four weeks. Efficacy of drugs was assessed by size reduction of the warts and HPV viral load quantification using droplet digital PCR. The gene expression profiling of CA was analyzed using NanoString Technology. After the podophyllin and C. nutans treatments, CA lesion sizes were reduced to 97.0% and 84.4% clearance, and the HPV viral loads were reduced by 74.0% and 46.6%, respectively. The gene expression pattern of immune profiling showed that 23 genes (i.e., HLA-DPB, CCL3, CXCL2, CXCR2, and OSM) were significantly differentially expressed by podophyllin, whereas 2 genes (IFNL1 and IRF2) were remarkably expressed by C. nutans. In inflammatory profiling, 108 genes (i.e., CXCL2, IL8, and STAT3) were highly expressed by podophyllin, but none of genes were observed to change expression by C. nutans. These results suggested that podophyllin may reduce the HPV infection through a mechanism related to proinflammatory response. In addition, C. nutans was found to suppress the HPV infection through mechanism related to the activation of immune response. This study shows novel therapeutic mechanisms of podophyllin and C. nutans. It is suggested that C. nutans might be used as an alternative treatment for CA treatment.

Clinacanthus nutans Lindau (Family: Acanthaceae) is a medicinal herb widely distributed in the tropic and subtropic areas of Asia. C. nutans is traditionally consumed as vegetable or herbal tea, as well as a folk medicine for anticancer and antifungal activities. However, to date, chemical constituent responsible for observed health beneficial effects of this medicinal plant is not clear. In the current study, 32 compounds (1−32), including three new megastigmanes (1–3) were isolated from the aerial parts of C. nutans. Their structures were elucidated on the basis of comprehensive NMR, MS, and CD spectroscopic data analysis, as well as chemical hydrolysis. Among the isolates, cycloartane triterpenoids (9, 10, and 12) displayed moderate anti-proliferative effects against HepG2 cell growth with IC50 values ranging from 9.12 to 19.89 μM. Data obtained from flow cytometry analysis and western blotting assays revealed that compounds 9 and 12 induced apoptosis of HepG2 cells by modulating the expression of proteins associated to mitochondrial-mediated apoptotic pathway. Furthermore, megastigmanes 1, 2, 7, and 8 enhanced the anti-Candida albicans activity of amphotericin B (AmB), supporting the synergistic effects between megastigmanes and AmB. This is the first report of anticancer and antifungal potential of cycloartane triterpenoids and megastigmanes in C. nutans, which shed useful insights on the relationship between C. nutans's chemical constituent and its beneficial effects to health. Findings from this study support further development of this medicinal plant for potential pharmaceutical applications.

Background Clinacanthus nutans extracts have been consumed by the cancer patients with the hope that the extracts can kill cancers more effectively than conventional chemotherapies. Our previous study reported its anti-inflammatory effects were caused by inhibiting Toll-like Receptor-4 (TLR-4) activation. However, we are unsure of its anticancer effect, and its interaction with existing chemotherapy. Methods We investigated the anti-proliferative efficacy of polar leaf extracts (LP), non-polar leaf extracts (LN), polar stem extract (SP) and non-polar stem extracts (SN) in human breast, colorectal, lung, endometrial, nasopharyngeal, and pancreatic cancer cells using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, MTT assay. The most potent extracts was tested along with gemcitabine using our established drug combination analysis. The effect of the combinatory treatment in apoptosis were quantified using enzyme-linked immunosorbent assay (ELISA), Annexin V assay, antibody array and immunoblotting. Statistical significance was analysed using one-way analysis of variance (ANOVA) and post hoc Dunnett’s test. A p-value of less than 0.05 (p Results All extracts tested were not able to induce potent anti-proliferative effects. However, it was found that pancreatic ductal adenocarcinoma, PDAC (AsPC1, BxPC3 and SW1990) were the cell lines most sensitive cell lines to SN extracts. This is the first report of C. nutans SN extracts acting in synergy with gemcitabine, the first line chemotherapy for pancreatic cancer, as compared to conventional monotherapy. In the presence of SN extracts, we can reduce the dose of gemcitabine 2.38–5.28 folds but still maintain the effects of gemcitabine in PDAC. SN extracts potentiated the killing of gemcitabine in PDAC by apoptosis. Bax was upregulated while bcl-2, cIAP-2, and XIAP levels were downregulated in SW1990 and BxPC3 cells treated with gemcitabine and SN extracts. The synergism was independent of TLR-4 expression in pancreatic cancer cells. Conclusion These results provide strong evidence of C. nutans extracts being inefficacious as monotherapy for cancer. Hence, it should not be used as a total substitution for any chemotherapy agents. However, SN extracts may synergise with gemcitabine in the anti-tumor mechanism.

The objective of the study is to analyse the phytochemical content by quantitative and qualitative methods and to investigate the antioxidant properties on Clinacanthus nutans leaves and stems from different areas: Yik Poh Ling (YPL) un-shaded sample (exposed to direct sunlight) and You Dun Chao (YDC) un-shaded and shaded (with black shade netting) samples. Extracts were prepared by dissolving fine powder of C. nutans in 1:10 solvent using ultrasonic extraction for an hour. Preliminary screening and quantitative estimation of phytochemical (alkaloids, flavonoids, cardiac glycosides, saponins, steroids, terpenoids, phenols and tannins) analysis were conducted. Then, determination of antioxidant properties (total phenolic content, TPC, ferric reducing/ antioxidant power, FRAP and radical-scavenging activities, DPPH) were conducted. All the samples contained alkaloids, flavonoids, cardiac glycosides, saponins, steroids, terpenoids, phenols and tannins. Un-shaded leaves of C. nutans from YDC exhibited significantly higher result in antioxidant properties (966.00mg GAE/g of dried sample in TPC, 20.44mg TE/g of dried sample in FRAP and 11.14mg TE/g of dried sample in DPPH) than the other samples (p < 0.05). As a conclusion, C. nutans has a good potential to be an alternative antioxidant source.

Clinacanthus nutans (Burm. f.) Lindau (C. nutans) has been reported to lower blood glucose level; however, evidence on its efficacy in lowering diabetic complications is limited. The antidiabetic properties of C. nutans aqueous leaf extract on serum metabolic indices, sorbitol production, and aldose reductase enzyme activities in the kidneys, lens, and sciatic nerve of type II diabetic (T2D) rats were evaluated. All rats except normal control rats were fed with a high‐fat diet for 8 weeks to induce obesity and subsequently injected with 35 mg/kg streptozotocin to induce type II diabetes. Aqueous leaf extract of C. nutans (100 and 200 mg kg−1 day−1) and quercetin (10 mg kg−1 day−1) were fed orally for 4 weeks. Diabetic rats administered with C. nutans at 100, 200 mg kg−1 day−1 and quercetin had significantly (p

The present study reported a simple method in synthesizing silver nanoparticles (AgNPs) by using Clinacanthus nutans extract. The UV–visible spectra showed the characteristic absorption peak at 480 nm, and the intensity was increased with the increase in plant extract ratio and incubation period. Analysis of particle size through FESEM, XRD and TEM revealed the average of synthesized AgNPs is 73.4 nm. The EDX analysis confirmed the formation of metallic nature of silver. The FTIR spectra indicated the role of carbonyl groups in the synthetic process and further confirmed by identification of flavonoids and their glycosides by UPLC-QTOF/MS. Antimicrobial activity of biosynthesized AgNPs shows effective inhibition against common bacterial strains including Bacillus subtilis, Enterococcus faecalis, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Proteus vulgaris.

Diabetes is an independent risk factor for the development of cardiovascular disease, with approximately 80% of cardiovascular mortality and morbidity linked to vascular complications such as atherosclerosis. It has been estimated that up to one-third of patients with diabetes mellitus use some form of complementary and/or alternative medicine. One plant that has received attention from diabetic patients for its perceived antidiabetic properties is Clinacanthus nutans, a member of the Acanthaceae family that is known as snake grass. Ethnomedical applications of this herb have been identified for the treatment of certain conditions, including fever, diabetes, skin rashes, and insect bites. This review aims to assess the potential of C. nutans to be used in the prevention and/or treatment of diabetic vasculopathy. Evidence for antidiabetic, anti-inflammatory, and dyslipidemic properties of C. nutans, as shown from experimental studies, is presented and discussed. Diabetes, inflammation, and hyperlipidemia are known to play significant roles in the initiation and severity of diabetic cardiovascular disease; thus, targeting these factors might be beneficial for preventing and/or treating diabetic vasculopathy.

Diabetes mellitus is a common metabolic disease marked by hyperglycemia. Diabetes also can be caused by oxidative stress. The administration of antioxidants, whether it’s from food or supplement can inhibit the formation of free radicals. Clinacanthus nutans is reported to possess various medicinal properties including blood glucose lowering effect, alpha-glucosidase inhibition activity, antioxidant activities, anti-cancer properties and anti-inflammatory effects. Based on research Clinacanthus nutans leaf extracts fractions has several shortcomings, amongst them are its viscosity that gave difficulties in formulation and its astringent and bitter taste that made it hard to use orally. To overcome these shortcomings, microencapsulation can be used. The purpose of this research is to find the blood glucose level lowering effect of the aqueous fraction of microcapsules and to find the effective dose on fructose and fat-induced Wistar strain rat. The induction using fructose and high-fat feed for 55 days was applied. Rats were divided into 8 groups: normal control (aquadest), negative control (Arabic gum and maltodextrin), positive control (metformin), dose I 15,89 mg/ kgBW (microcapsule of aqueous fraction of Clinacanthus nutans extract), dose II 31,78 mg/ kgBW, dose III 47,67 mg/ kgBW, dose IV 63.56 mg /kgBW, and dose V 79.45 mg/ kgBW. The result showed that the microcapsules of the aqueous fraction of Clinacanthus nutans (Burm.f) extract could lower Wistar strain rat blood glucose level in the dose of 15.89 mg/kg of the rat’s body weight.

Clinacanthus nutans Lindau has been traditionally used for healing of bone fragility, but the mechanism of actions has not been clarified yet. In this study, the bone regeneration activity of lupeol derived from C. nutans was assessed using an in vitro model of osteoblast cells MC3T3-E1. The finding revealed that the compound was not significantly toxic to osteoblast cells at concentration of ≤40 μg/mL. Lupeol demonstrated the osteogenic activity through enhancement of alkaline phosphatase (ALP) of osteoblast cells up to 31.2%, 21%, and 12% at concentrations of 5, 10, and 20 µg/mL, respectively ( p p p

Triple-negative breast cancer is the main type of breast carcinoma that causes mortality among women because of the limited treatment options and high recurrence. Chronic inflammation has been linked with the tumor microenvironment (TME) in breast cancer progression. Clinacanthus nutans (CN) has gained much attention because of its anticancer properties, but its mechanism remains unclear. We aimed to study the qualitative phytochemical content and elucidate the cytotoxicity effects of CN on human triple-negative breast cancer (TNBC), MDA-MB-231 and human macrophage-like cells such as THP-1 by using sulforhodamine B (SRB) assay. As highly metastatic cells, MDA-MB-231 cells can migrate to the distal position, the effect of CN on migration were also elucidated using the scratch assay. The CN effects on ameliorating chronic inflammation in TME were studied following the co-culture of MDA-MB-231/THP-1 macrophages. The cytokine expression levels of IL-6, IL-1β and tumor necrosis factor-alpha (TNF-α) were determined using ELISA assays. The results showed that both ethanolic and aqueous CN extracts contained alkaloid, phenol and tannin, flavonoid, terpenoid, glycoside and steroid. However, saponin was only found in the aqueous extract of CN. CN was not cytotoxic to both MDA-MB-231 and THP-1 cells. The ability of MDA-MB-231 to migrate was also not halted by CN treatment. However, CN ethanol extract decreased IL-6 at 25 μg/mL (p = 0.02) and 100 μg/mL (p = 0.03) but CN aqueous extract increased IL-6 expression at 50 μg/mL (p = 0.08) and 100 μg/mL (p = 0.02). IL-1β showed decreased expression after treated with CN ethanol and CN aqueous both at 25 μg/mL (p = 0.03). TNF-α were significantly decreased after CN ethanol treatment at concentration 25- (p = 0.001), 50- (p = 0.000) and 100 μg/mL (p = 0.000). CN aqueous extract slightly inhibited TNF-α at all 25–50- and 100 μg/mL (p = 0.001, p = 0.000, p = 0.000, respectively). Overall, CN acts by ameliorating the pro-inflammatory condition in the TME and may be a potential strategy for its anticancer mechanism on highly metastatic breast cancer condition. The major pathways that link both cancer and inflammation were NF-κB and STATs thus further study on the upstream and downstream pathways is needed to fully understand the mechanism of CN extracts in cooling the inflamed TME in breast cancer.

This appraisal is comprised of the inflammatory studies that have been conducted on Clinacanthus nutans, Acanthus ebracteatus, and Barleria lupulina. The review aims to provide a comprehensive evaluation of the supporting and contradictory evidence on each plants' anti-inflammatory properties, whilst addressing the gaps in the current literature. The databases used to obtain relevant studies were Google Scholar, ResearchGate, PubMed and Nusearch (University of Nottingham). A total of 13 articles were selected for this review. A. ebracteatus was found to suppress neutrophil migration and weakly inhibits chronic inflammatory cytokines. Furthermore, B. lupulina and C. nutans were shown to possess very similar anti-inflammatory properties. The studies on C. nutans indicated that its anti-inflammatory effect is strongly related to the inhibition of toll-like receptor 4 (TLR4). Moreover, several phytoconstituents isolated from B. lupulina were shown to activate the anti-inflammatory Nrf2 pathway. Overall, all the studies have provided evidence to support the use of these plants as anti-inflammatory herbal remedies. However, their exact mechanism of action and the responsible phytoconstituents are yet to be established.

Clinacanthus nutans, commonly known as Sabah snake grass, is one of the more important medicinal plants in Malaysia&rsquo, s herbal industry. C. nutans has gained the attention of medical practitioners due to its wide range of bioactive compounds responsible for various biological activities, such as anti-cancer, anti-venom and anti-viral activities. Due to its high pharmacological properties, the species has been overexploited to meet the demands of the pharmaceutical industry. The present study was conducted to establish a suitable in vitro culture procedure for the mass propagation of C. nutans. Murashige and Skoog (MS) basal medium, supplemented with different types of cytokinins, auxins, basal medium strength and sucrose concentrations, were tested. Based on the results, a full-strength MS basal medium supplemented with 12 &micro, M 6-benzylaminopurine (BAP) and 30 g/L sucrose was recorded as the best outcome for all the parameters measured including the regeneration percentage, number of shoots, length of shoots, number of leaves and fresh weight of leaves. In the analysis of the phenolics content and antioxidant activities, tissue-cultured leaf extracts assayed at 100 &deg, C exhibited the highest phenolic content and antioxidant activities. The propagation of C. nutans via a plant tissue culture technique was recorded to be able to produce high phenolic contents as well as exhibit high antioxidant activities.

Sabah snake grass or Clinacanthus nutans has drawn public interest having significant economic benefits attributable to the presence of phytochemicals and several interesting bioactive constituents that may differ according to harvesting age and harvesting frequency. The current study was aimed to evaluate the effect of harvesting age and harvesting frequency towards herbal yield, antioxidant activities, phytochemicals synthesis, and bioactive compounds of C. nutans. A factorial randomized completely block design with five replications was used to illustrate the relationship between herbal yield, DPPH (2, 2-diphenyl-1-picrylhydrazyl) and ferric reducing antioxidant power (FRAP) assays, total phenolic and flavonoid content affected by harvesting age (week 8, 12, and 16 after transplanting), and harvesting frequency (harvest 1, 2, and 3). The bioactive compounds by HPLC were also determined to describe the interaction effect between both harvesting age and harvesting frequency. The yield, antioxidant activities, and phytochemical contents were gradually increased as the plant grew, with the highest recorded during week 16. However, the synthesis and activities of phytochemicals were reduced in subsequent harvests despite the increment of the herbal yield. All bioactive compounds were found to be influenced insignificantly and significantly by harvesting age and harvesting frequency, respectively, specifically to shaftoside, iso-orientin, and orientin. Among all constituents, shaftoside was the main compound at various harvesting ages and harvesting frequencies. These results indicated that harvesting at week 16 with 1st harvest frequency might enhance the yield while sustaining the high synthesis of polyphenols and antioxidant activities of C. nutans.

Medicinal plants are commonly used in complementary and alternative medicine to treat several chronic diseases. Gynura procumbens, Hibiscus sabdariffa and Clinacanthus nutans are believed to contain high level of antioxidants including phenolic acids that may contribute to antioxidant activities. Thus, the objectives of this study were to determine total phenolic content (TPC) and antioxidant scavenging capacity of these plants using three different extracts as well as to determine the relationship between TPC and antioxidant activity. The quantification of phenolic compound was conducted by analyzing the compounds at three different fractions (free, insoluble bound and soluble bound). Three replicates of the medicinal plants were extracted with 80% methanol (free fraction) and undergone hydrolysis with HCl (soluble bound) and NaOH (insoluble bound). The TPC was determined using Folin-Ciocalteau reagent while antioxidant radical scavenging capacities were analyzed using DPPH assay. Results showed that G. procumbens and C. nutans of the insoluble bound extracts had the highest amount of TPC compared to the free phenolic and soluble bound (p

Clinacanthus nutans is a well-known herb that has been used as an alternative and therapeutic medicine, however more selective C. nutans extracts are needed. In this study, leaves were extracted with 80% methanol and further fractionated with n-hexane, dichloromethane, chloroform, n-butanol, and aqueous residue. Subsequently, the total phenolic content (TPC), total flavonoid content (TFC), antioxidant scavenging activity, and antiproliferative effects on breast cancer (Michigan Cancer Foundation-7 [MCF7]) and normal breast (Michigan Cancer Foundation-10A [MCF 10A]) cells of the extracts were measured. Additionally, molecular docking simulation of the major compounds from C. nutans extracts was conducted. The aqueous residue had the highest TPC and TFC, whereas the crude extract had the highest scavenging activity. Among the extracts, dichloromethane extract (CN-Dcm) was selected as it had the highest selectivity index (SI) (1.48). Then, the chosen extract (CN-Dcm) was proceed for further analysis. The compounds from CN-Dcm were identified using gas chromatography&ndash, mass spectrometry (GC&ndash, MS). The major compounds from CN-Dcm were further investigated through molecular docking studies. Palmitic acid and linolenyl alcohol were the compounds found in the CN-Dcm extract that exhibited the highest binding affinities with p53-binding protein Mdm-2. These results highlight the potential of C. nutans as a source of anticancer activities.

Background: Scientific support for Thai traditional medicine (TTM) practice is warranted for reintroduction into modern healthcare systems. A promising TTM practice for treatment of pressure ulcers was selected to conduct a clinical trial. This study aimed to evaluate the efficacy of the TTM practice for the treatment of pressure ulcers using honey or a Thai Herbal Oil preparation (THO) based on the TTM wound diagnosis comparing with the standard practice. Methods: The study design was an open-label randomized controlled trial. Sixty-six participants, with pressure ulcers at least stage II-IV or unstageable, were allocated to two groups via minimization. A TTM practice group received honey or THO depending on the TTM diagnosis via the Thai Traditional Medicine Pressure Ulcer Assessment Tool (TTM-PUAT). A standard practice group received advanced dressings, including hydrogel, alginate, silver-impregnated, or hydrocolloid dressings. The primary outcome was the Pressure Ulcer Scale for Healing (PUSH). Results: Both TTM practice and standard practice showed a significant reduction in PUSH scores after treatments. However, there was no significant difference in PUSH score reduction between the groups. The mean PUSH score reduction over the 6-week period was 2.58 ± 3.38 (95% CI 1.34–3.82) in the TTM practice group and 3.24 ± 3.49 (95% CI 1.91–4.57) in the standard practice group (p = 0.284). The TTM practice and standard practice accelerated pressure ulcer healing without statistically significant difference between the practices, during 6 weeks in a home-based care setting. This finding supported the TTM practice as an alternative treatment for pressure ulcer. Keywords: Decubitus, Diagnosis, Honey, Clinacanthus nutans, Zingiber montanum, Traditional medicine

The purpose of this study was to investigate apoptotic activity of silver nanoparticle Clinacanthus nutans (AgNps-CN) towards HSC-4 cell lines (oral squamous cell carcinoma cell lines).Methods involved were MTT assay (cytotoxic activity), morphological cells analysis, flow cytometry and cell cycle analysis and western blot.MTT assay revealed ICAgNps-CN have shown potential in inhibiting HSC-4 cell lines. IC

Cyprinid herpesvirus 3 (CyHV-3) or koi herpesvirus (KHV) is a virulent viral infection in common carp and koi. The disease has caused global epizootic and economic loss in fish aquaculture and in the wild. Clinacanthus nutans (Burm. f.) Lindau is a well-known medicinal plant used in Thai traditional medicine. Virucidal effects of the plant extract against human herpes simplex virus have been reported. In this study, C. nutans crude extract was tested for antiviral activities against CyHV-3 in koi carp. Results showed effective antiviral activity against CyHV-3 pre- and post-infection. The 50% lethal concentration (LC50 ) of extract was higher than 5 mg/ml. The 50% effective dose (ED50 ) was 0.99 mg/ml, 0.78 mg/ml, 0.75 mg/ml and 0.71 mg/ml at 1, 2, 3 and 4 hr pre-infection, respectively. The ED50 from post-infection tests was 2.05 mg/ml and 2.34 mg/ml at 0 and 24 hr, respectively. These results demonstrated that crude extract expressed antiviral activity against CyHV-3 and can be applied as a therapeutic agent in common carp and koi aquaculture.

Traditionally, Clinacanthus nutans (CN) or locally named as ‘Belalai Gajah’ is one of the herbal plant claimed to be able to treat cancer. The aimd of this study are to extract, isolate and characterize the active anticancer compound from CN and to determine the mode of cell death induced by the compound. Bioassay guided fractionation was done on the CN extract by using column chromatography. The cytotoxicity activities of these fractions toward HeLa cells were examined by MTT assay. The nuclear morphology was examined by Hoechst 33258 staining and the cell cycle arrest was evaluated by propium iodide staining using flow cytometry. The presence of active compound in the chosen fraction was determined by Liquid Chromatography Mass Spectrometry (LCMS). Out of 16 fractions collected, Fraction 11(F11) showed the lowest IC 50 value with 27± 2.6µg/mL. The value of IC 50 for F11 towards normal cell, NIH 3T3 cell and L929 cell, were 70 ± 4.0µg/mL and 45 ± 1.5µg/mL respectively. These values were higher than tamoxifen, therefore indicating that tamoxifen is more toxic towards normal cells compared to F11. Nuclear morphology of HeLa cell displayed DNA fragmentation, nuclear condensation and formation of apoptotic bodies upon treatment with F11 for 24 hours. The cell cycle distribution of HeLa cell treated with F11 was arrested at G1 phase. The active compound identified to potentially possess the anticancer property is 19-Oxo-all-trans-retinoic acid. In conclusion, 19-Oxo-all-trans-retinoic acids from F11 of the CN extract, is a potential anticancer agent for cervical cancer. DOI : http://dx.doi.org./10.17576/JSKM-2018-1602-06

Clinacanthus nutans (C. nutans) leaf extracts have been widely used by cancer patients in Malaysia and local practice claims a cure to cancer. There were several studies done to determine the cytotoxicity potency of C. nutans extracts on various types of cells. However, there is still lacking on the knowledge regarding the combination effect of C. nutans with anticancer drugs. Thus, the study was carried out to determine the cytotoxicity potency of C. nutans extracts and paclitaxel (PTX) alone and, in combination on MDA-MB-231 cells. The cells were treated with 100% ethanol extract of C. nutans (CNE) and water extract of C. nutans (CNA), PTX and combination of both extracts and PTX for 72 hours and the cytotoxic activity was determined using SRB assay. Result showed that CNE had little cytotoxic activity, whereas CNA showed no cytotoxic activity on MDA-MB-231 cells. For combination treatment of C. nutans extracts and PTX, only CNE showed significant enhanced PTX-induced cytotoxicity (p < 0.05), meanwhile CNA inhibited PTX-induced cytotoxicity significantly (p < 0.05). As a conclusion, CNE was able to increase PTX potency to inhibit the viability of MDA-MB-231 cells. DOI : http://dx.doi.org./10.17576/JSKM-2018-14

Clinacanthus nutans (Burm. f.) Lindau is a traditional medicinal plant belonging to the Acanthaceae family. Its therapeutic potentials have been increasingly documented particularly the antiviral activity against Herpes Simplex Virus (HSV), anti-cancer, anti-oxidant, anti-inflammatory and immunomodulatory activities. However, majority of these studies used crude or fractionated extracts and not much is known about individual compounds from these extracts and their biological activities. In the present study, we have isolated four compounds (CN1, CN2, CN3 and CN4) from the hexane fractions of C. nutans leaves. Using NMR spectroscopic analysis, these compounds were identified to be shaftoside (CN1), stigmasterol (CN2), β-sitosterol (CN3) and a triterpenoid lupeol (CN4). To determine the immunosuppressive potential of these compounds, their effects on mitogens induced T and B lymphocyte proliferation and the secretion of helper T cell cytokines were examined. Among the four compounds, stigmasterol (CN2) and β-sitosterol (CN3) were shown to readily inhibit T cell proliferation mediated by Concanavalin A (ConA). However, only β-sitosterol (CN3) and not stigmasterol (CN2) blocks the secretion of T helper 2 (Th2) cytokines (IL-4 and IL-10). Both compounds have no effect on the secretion of Th1 cytokines (IL-2 and IFN-γ), suggesting that β-sitosterol treatment selectively suppresses Th2 activity and promotes a Th1 bias. CN3 was also found to significantly reduce the proliferation of both T helper cells (CD4+CD25+) and cytotoxic T cells (CD8+CD25+) following T cell activation induced by ConA. These results suggested that phytosterols isolated from C. nutans possess immunomodulatory effects with potential development as immunotherapeutics.

Subcritical carbon dioxide Soxhlet extraction of biologically active compounds from Clincanthus nutans was investigated by full factorial design to identify and optimize the factors (particle size and co-solvent) affecting extract yield, antioxidant activity, total phenolic content, total flavonoid content, and α-glucosidase inhibitory activity. An average of 3.103% yield, 98.90% antioxidant activity, 49.40 mg/g (GAE) TPC, 43.76 mg/g (RE), and 88.58% AGI activity can be achieved using the optimum levels of independent variables. The GC-Q-TOF MS identification of optimized extract shown that different classes of phytoconstituents were successfully separated by CO2-Soxhlet to produce potential antioxidant and α-glucosidase inhibitory activity.

Plants are believed to have various medicinal benefits from their constituents, but they might also give toxic effects to the human's body. Clinacanthus nutans that belongs to Acanthaceae family, which has been reported to have an antidiabetic and antioxidant effect. The study has been done to determine the toxic level and the lethal concentration of non-polar extract of C. nutans leaves using zebrafish embryos. Crude extract of 80% methanolic solution was further partitioned to obtain the n-hexane fraction. Upon spawning and fertilization, the eggs were collected, transferred into a 96-well plate, and incubated with the n-hexane fraction at different concentrations ranging from 15.63-500 µg/ml prepared in 2% DMSO. The median lethal concentration (LC50) was calculated to be 75.49 µg/mL using probit analysis. The reading is considered toxic to the embryos. The heartbeat rate was measured using DanioScope software, and the mortality and survival rate were assessed by microscopic observation at 24, 48, 72, and 96 hours post-fertilization. Some of the morphological defects observed include less pigmentation, crooked backbone, dented tail, and oedema. The n-hexane fraction's profiling using gas chromatography-mass spectroscopy (GC-MS) analysis showed the presence of some fatty acids, organic acid, and sterols, which is known to be toxic in synergism.

Clinacanthus nutans (CN) (Acanthaceae) is well-known for its anti-inflammatory properties among Asian communities; however, there are currently no data specifically focused on the anti-inflammatory effects of CN on the brain tissue. Neuroinflammation is a common consequence of toxin intrusion to any part of the central nervous system (CNS). As an innate immune response, the CNS may react through both protective and/or toxic actions due to the activation of neuron cells producing pro- and/or anti-inflammatory cytokines in the brain. The unresolved activation of the inflammatory cytokines' response is associated with the pathogenesis of neurological disorders. The present study aimed to decipher the metabolic mechanism on the effects of 14 days oral treatment with CN aqueous extract in induced-lipopolysaccharides (LPS) rats through 1H NMR spectroscopic biomarker profiling of the brain tissue and the related cytokines. Based on the principal component analysis (PCA) of the nuclear magnetic resonance (NMR) spectral data, twenty-one metabolites in the brain tissue were profiled as biomarkers for the LPS (10 μL)-induced neuroinflammation following intracerebroventricular injection. Among the twenty-one biomarkers in the neuroinflammed rats, CN treatment of 1000 and 500 mg/kg BW successfully altered lactate, pyruvate, phosphorylcholine, glutamine, and α-ketoglutarate when compared to the negative control. Likewise, statistical isolinear multiple component analysis (SIMCA) showed that treatments by CN and the positive control drug, dextromethorphan (DXM, 5 mg/kg BW), have anti-neuroinflammatory potential. A moderate correlation, in the orthogonal partial least squares (OPLS) regression model, was found between the spectral metabolite profile and the cytokine levels. The current study revealed the existence of high levels of pro-inflammatory cytokines, namely IL-1α, IL-1β, and TNF-α in LPS-induced rats. Both CN dose treatments lowered IL-1β significantly better than DXM Interestingly, DXM and CN treatments both exhibited the upregulation of the anti-inflammatory cytokines IL-2 and 4. However, DXM has an advantage over CN in that the former also increased the expression of IL-10 of anti-inflammatory cytokines. In this study, a metabolomics approach was successfully applied to discover the mechanistic role of CN in controlling the neuroinflammatory conditions through the modulation of complex metabolite interactions in the rat brain.

The present work showed the preparation of gold (Au NPs) and silver nanoparticles (Ag NPs) using aqueous leaf extract of Clinacanthus nutans Lindau (C. Lindau). The prepared NPs were studied using various microscopic and spectroscopic techniques, which confirmed the formation of crystalline Ag NPs, Au NPs that are stabilized with C. Lindau extract polyphenols. The prepared Au NPs and Ag NPs are studied to assess their comparative analgesic and muscle relaxant activities conducted on BALB/c mice. The muscle relaxant studies displayed that Ag NPs were comparatively higher efficient than Au NPs and methanolic C. Lindau extract in traction examination. Additionally, the analgesic studies exhibited that Ag NPs, Au NPs showed maximum percentage reduction in acetic acid induced writhing at the concentrations of 50, 100 and 150 mg/kg body weight. Further, these results conclude that as prepared Au NPs and Ag NPs prepared from C. Lindau leaf extract demonstrated very good analgesic and muscle relaxant activities for the use of pain management in nursing care.

Background Methanol extract (MECN) of Clinacanthus nutans Lindau leaves (family Acanthaceae) demonstrated peripherally and centrally mediated antinociceptive activity via the modulation of opioid/NO-mediated, but cGMP-independent pathway. In the present study, MECN was sequentially partitioned to obtain petroleum ether extract of C. nutans (PECN), which was subjected to antinociceptive study with aims of establishing its antinociceptive potential and determining the role of opioid receptors and l–arginine/nitric oxide/cyclic-guanosine monophosphate (l–arg/NO/cGMP) pathway in the observed antinociceptive activity. Methods The antinociceptive potential of orally administered PECN (100, 250, 500 mg/kg) was studied using the abdominal constriction-, hot plate- and formalin-induced paw licking-test in mice (n = 6). The effect of PECN on locomotor activity was also evaluated using the rota rod assay. The role of opioid receptors was determined by pre-challenging 500 mg/kg PECN (p.o.) with antagonist of opioid receptor subtypes, namely β–funaltrexamine (β–FNA; 10 mg/kg; a μ-opioid antagonist), naltrindole (NALT; 1 mg/kg; a δ-opioid antagonist) or nor–binaltorphimine (nor–BNI; 1 mg/kg; a κ-opioid antagonist) followed by subjection to the abdominal constriction test. In addition, the role of l–arg/NO/cGMP pathway was determined by prechallenging 500 mg/kg PECN (p.o.) with l–arg (20 mg/kg; a NO precursor), 1H-[1, 2, 4] oxadiazolo [4,3-a]quinoxalin-1-one (ODQ; 2 mg/kg; a specific soluble guanylyl cyclase inhibitor), or the combinations thereof (l–arg + ODQ) for 5 mins before subjection to the abdominal constriction test. PECN was also subjected to phytoconstituents analyses. Results PECN significantly (p 0.05) affect the locomotor activity of treated mice. The antinociceptive activity of PECN was significantly (p 0.05) affected by ODQ. HPLC analysis revealed the presence of at least cinnamic acid in PECN. Conclusion PECN exerted antinocicpetive activity at peripheral and central levels possibly via the activation of non-selective opioid receptors and modulation of the NO-mediated/cGMP-independent pathway partly via the synergistic action of phenolic compounds.

Sabah Snake Grass (Clinacanthus nutans) is an interesting traditional medicine because of pharmacology characteristics that contain rich in phenolic content. The aim of the study was to determine the optimum flavonoid compound on the antioxidant activities. The optimum extraction parameters for optimum recovery of phenolic content from Clinachantus nutans leaves were determined using response surface methodology (RSM). A central composite rotatable design (CCRD) was employed to investigate the effects of ultrasonic frequency (X1: 25 - 40 kHz), temperature (X2: 40 - 80 °C) and solid-to-solvent ratio (X3: 10 - 30 ml/g) on the recovery of flavonoid vitexin (Y1) and orientin (Y2). The optimal parameter achieved based on combination of the extraction parameters are as follows: X1 = 25 kHz, X2 = 40 °C and X3 = 15 g/ml. These optimum conditions yielded vitexin (2.25 mg/g) and orientin (3.75 mg/g). Our results revealed that distilled water may be a good choice for extracting the phenolic compound of Clinacanthus nutans due to costing and environmental issue. Vitexin and orientin have potential of antiproliferate activity on the cancer cell. The concentration of extracted compound (orientin and vitexin) from the optimum parameters had produced high in phenolic content. In conclusion, the application of response surface method successfully provide the high extraction parameters with high total phenolic content and antioxidant activity.

Methanolic extract of Clinacanthus nutans Lindau leaves (MECN) has been reported to exert antinociceptive activity. The present study aimed to elucidate the possible antinociceptive mechanisms of a lipid-soluble fraction of MECN, which was obtained after sequential extraction in petroleum ether. The petroleum ether fraction of C. nutans (PECN), administered orally to mice, was (i) subjected to capsaicin-, glutamate-, phorbol 12-myristate 13-acetate-, bradykinin-induced nociception model; (ii) prechallenged (intraperitoneal (i.p.)) with 0.15 mg/kg yohimbine, 1 mg/kg pindolol, 3 mg/kg caffeine, 0.2 mg/kg haloperidol, or 10 mg/kg atropine, which were the respective antagonist of α2-adrenergic, β-adrenergic, adenosinergic, dopaminergic, or muscarinic receptors; and (iii) prechallenged (i.p.) with 10 mg/kg glibenclamide, 0.04 mg/kg apamin, 0.02 mg/kg charybdotoxin, or 4 mg/kg tetraethylammonium chloride, which were the respective inhibitor of ATP sensitive-, small conductance Ca2+-activated-, large conductance Ca2+-activated-, or nonselective voltage-activated-K+ channel. Results obtained demonstrated that PECN (100, 250, and 500 mg/kg) significantly (P+ channel blockers. However, only pretreatment with apamin and charybdotoxin caused full inhibition of PECN-induced antinociception. The rest of the K+ channel blockers and all antagonists caused only partial inhibition of PECN antinociception, respectively. Analyses on PECN’s phytoconstituents revealed the presence of antinociceptive-bearing bioactive compounds of volatile (i.e., derivatives of γ–tocopherol, α–tocopherol, and lupeol) and nonvolatile (i.e., cinnamic acid) nature. In conclusion, PECN exerts a non-opioid-mediated antinociceptive activity involving mainly activation of adenosinergic and cholinergic receptors or small- and large-conductance Ca2+-activated-K+ channels.

Background: Clinacanthus nutans exhibits antioxidant, anti-inflammatory, antiviral, anticancer, and anti-biofilm activities. Wounds in the oral cavity can affect the quality of life. Wound healing is a complex process to restore functions of injured tissues. The effect of C. nutans on oral mucosal burns and tongue wounds of rats is investigated. Methods: Forty male Sprague Dawley rats were divided into negative control and treatment groups with 25, 50, and 100 mg/mL of C. nutans. Oral mucosal burns and tongue wounds were observed on days 7 and 14 based on the fibroblast count and collagen deposits. Data were analyzed by analysis of variance and post hoc Tukey’s test with P < 0.05. Results: Results revealed that on day 7, the fibroblast count of mucosal burns and tongue wounds increases. The maximum mucosal burns were observed for the 100 mg/mL group, whereas the maximum tongue wounds were observed for the 25 mg/mL group. On day 7, collagen deposits were increased in case of mucosal burns. The number of collagen deposits increased on day 14, the highest mucosal burns were observed for the 100 mg/mL group, and in case of tongue wounds, the highest number was observed for the 50 mg/mL group. Conclusion: Ethanol extracts of C. nutans leaves can increase the fibroblast count on day 7 and collagen deposits on day 14 after injury in case of oral mucosal burns and tongue wounds during the wound healing of Sprague Dawley rats.

Breast cancer is one of the most common cancers with a relatively high mortality rate. Despite the advancement of its medical treatments, many patients are still seeking complementary alternative medicines, namely Clinacanthus nutans which is found mainly in South-East Asian countries. We aim to find the antioxidant properties and cytotoxic activity of the plant extract toward breast cancer cell lines Michigan Cancer Foundation-7 (MCF7) and T47D individually and in combination with doxorubicin. Extractions of C. nutans with ethanol, n-hexane, and ethyl acetate were done using rotatory vacuum evaporators with the reflux method. Screening of biochemical properties was conducted. Antioxidant activity was measured toward α, α-diphenyl-β-picrylhydrazyl (DPPH) with IC50 scores were shown to be highest in ethyl acetate extract. Cytotoxic effects of all three extracts were shown to be low in both MCF7 and T47D cells. However, combinations of 125 μg/ml n-hexane extract of C. nutans, and 0.1 μg/ml doxorubicin in T47D cancer cells showed further proliferation reduction compared to the single administration. The results suggested possible synergisms of the treatment combination.

Clinacanthus nutans has been used in traditional herbal medicine for cancer prevention, but the specific bioactive compounds responsible for the observed activities have not been explored. Different polar solvents such as methanol, chloroform, ethyl acetate, and hexane were used for the extraction. The extracts, fractions, and isolated compounds were subjected to DPPH and ferric reducing antioxidant potential (FRAP) assays. Methanol extracts show significant free-radical scavenging activity of 69.09% in DPPH and 56.49% FRAP. Purification of MeOH extracts afforded the fraction FB28 and two new sulfur-containing compounds, named clinamide D and E (1, 2). Compound (1) proved to be more active with an IC50 value for DPPH radical scavenging of 118.27 ± 0.01 µg/mL and reduction of Fe3+–TPTZ complex of 386.24 ± 0.02, higher than that of the standard ascorbic acid. Sulfur-containing compounds isolated from C. nutans is a potential natural antioxidant.

Clinacanthus nutans Lindau (C. nutans), commonly known as Sabah Snake Grass in southeast Asia, is widely used in folk medicine due to its analgesic, antiviral, and anti-inflammatory properties. Our recent study provided evidence for the regulation of cytosolic phospholipase A2 (cPLA2) mRNA expression by epigenetic factors (Tan et al. in Mol Neurobiol. doi: 10.1007/s12035-015-9314-z , 2015). This enzyme catalyzes the release of arachidonic acid from glycerophospholipids, and formation of pro-inflammatory eicosanoids or toxic lipid peroxidation products such as 4-hydroxynonenal. In this study, we examined the effects of C. nutans ethanol leaf extracts on epigenetic regulation of cPLA2 mRNA expression in SH-SY5Y human neuroblastoma cells and mouse primary cortical neurons. C. nutans modulated induction of cPLA2 expression in SH-SY5Y cells by histone deacetylase (HDAC) inhibitors, MS-275, MC-1568, and TSA. C. nutans extracts also inhibited histone acetylase (HAT) activity. Levels of cPLA2 mRNA expression were increased in primary cortical neurons subjected to 0.5-h oxygen-glucose deprivation injury (OGD). This increase was significantly inhibited by C. nutans treatment. Treatment of primary neurons with the HDAC inhibitor MS-275 augmented OGD-induced cPLA2 mRNA expression, and this increase was modulated by C. nutans extracts. OGD-stimulated increase in cPLA2 mRNA expression was also reduced by a Tip60 HAT inhibitor, NU9056. In view of a key role of cPLA2 in the production of pro-inflammatory eicosanoids and free radical damage, and the fact that epigenetic effects on genes are often long-lasting, results suggest a role for C. nutans and phytochemicals to inhibit the production of arachidonic acid-derived pro-inflammatory eicosanoids and chronic inflammation, through epigenetic regulation of cPLA2 expression.

Many population-based epidemiological studies have unveiled an inverse correlation between intake of herbal plants and incidence of stroke. C. nutans is a traditional herbal medicine widely used for snake bite, viral infection and cancer in Asian countries. However, its role in protecting stroke damage remains to be studied. Despite of growing evidence to support epigenetic regulation in the pathogenesis and recovery of stroke, a clear understanding of the underlying molecular mechanisms is still lacking. In the present study, primary cortical neurons were subjected to in vitro oxygen-glucose deprivation (OGD)-reoxygenation and hypoxic neuronal death was used to investigate the interaction between C. nutans and histone deacetylases (HDACs). Using pharmacological agents (HDAC inhibitor/activator), loss-of-function (HDAC siRNA) and gain-of-function (HDAC plasmid) approaches, we demonstrated an early induction of HDAC1/2/3/8 and HDAC6 in neurons after OGD insult. C. nutans extract selectively inhibited HDAC1 and HDAC6 expression and attenuated neuronal death. Results of reporter analysis further revealed that C. nutans suppressed HDAC1 and HDAC6 transcription. Besides ameliorating neuronal death, C. nutans also protected astrocytes and endothelial cells from hypoxic-induced cell death. In summary, results support ability for C. nutans to suppress post-hypoxic HDACs activation and mitigate against OGD-induced neuronal death. This study further opens a new avenue for the use of herbal medicines to regulate epigenetic control of brain injury.

Clinacanthus nutans (CN) is used traditionally for treating various illnesses. Robust safety data to support its use is lacking.To evaluate the adverse effects of aqueous extract of CN leaves (AECNL).The oral toxicity of the AECNL was tested following Organisation for Economic Co-operation and Development (OECD) guidelines. Mutagenicity (Ames test) of AECNL was evaluated using TA98 and TA100 Salmonella typhimurium strains.No mortality or morbidity was found in the animals upon single and repeated dose administration. However, significant body weight loss was observed at 2000 mg/kg during sub-chronic (90 d) exposure. In addition, increased eosinophil at 500 mg/kg and decreased serum alkaline phosphatase levels at 2000 mg/kg were observed in male rats. Variations in glucose and lipid profiles in treated groups were also observed compared to control. Ames test revealed no evidence of mutagenic or carcinogenic effects at 500 μg/well of AECNL.The median lethal dose (LD50) of the AECNL is5000 mg/kg and the no-observed-adverse-effect level is identified to be greater than 2000 mg/kg/day in 90-d study.

Objective To evaluate the monogalactosyl diglyceride (MGDG) and digalactosyl diglyceride (DGDG) from Clinacanthus nutans ( C. nutans ) for their in vitro antiviral activities against herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) by plaque reduction assay. Methods MGDG and DGDG were extracted with chloroform from C. nutans leaves. MGDG and DGDG were separated from chloroform crude extract using column chromatography, characterized by thin layer chromatography and quantified by high performance liquid chromatography. The anti HSV-1 and 2 activity against pre-treatment and post-treatment of the compounds was evaluated using plaque reduction assay. The cytotoxicity of the extract and the compounds on Vero cells were performed by MTT assay. Results MGDG and DGDG obtained by column chromatography showed identical profiles as standard MGDG and standard DGDG using thin layer chromatography and high performance liquid chromatography. MGDG and DGDG from C. nutans showed 100% inhibition of HSV-1 replication at the post step of infection at noncytotoxic concentration with IC 50 values of 36.00 and 40.00 μg/mL, and HSV-2 at 41.00 and 43.20 μg/mL, respectively. Moreover, MGDG and DGDG from C. nutans were demonstrated to have anti-herpes simplex activity at the same level as standard synthetic compounds. In contrast, pre-treatment of Vero cells with MGDG and DGDG before HSV-1 and HSV-2 infection did not show inhibitory effect against these viruses. MGDG and DGDG exhibited antiviral activity against HSV-1 with selectivity index of 26.00 and 23.00 and HSV-2 of 23.30 and 21.30. Conclusions MGDG and DGDG from C. nutans , a traditional Thai herbal medicine illustrated inhibitory activity against HSV-1 and HSV-2, probably by inhibiting the late stage of multiplication, suggesting their promising use as anti-HSV agents.

The objectives of the present study were to determine the mechanisms of antinociceptive effect of methanol extract ofClinacanthus nutans(Acanthaceae) leaves (MECN) using various animal nociceptive models. The antinociceptive activity of orally administered 10% DMSO, 100 mg/kg acetylsalicylic acid (ASA), 5 mg/kg morphine, or MECN (100, 250, and 500 mg/kg) was determined using the acetic acid-induced abdominal constriction (ACT), formalin-induced paw licking (FT), and hot plate tests (HPT). The role of opioid and nitric oxide/cyclic guanosine monophosphate (NO/cGMP) systems was also investigated. The results showed that MECN produced a significant (p<0.05) antinociceptive response in all nociceptive models with the recordedED50value of 279.3 mg/kg for the ACT, while, for the early and late phases of the FT, the value was >500 mg/kg or 227.7 mg/kg, respectively. This antinociceptive activity was fully antagonized by naloxone (a nonselective opioid antagonist) but was partially reversed byL-arginine (L-arg; a nitric oxide [NO] precursor), Nω-nitro-L-arginine methyl ester hydrochloride (L-NAME; an NO synthase inhibitor), or their combinations thereof. In contrast, 1H-[1,2,4]oxadiazole[4,3-a]quinoxalin-1-one (ODQ; a soluble guanylyl cyclase inhibitor) enhanced the extract’s antinociception. UHPLC analysis revealed the presence of several flavonoid-based compounds with antinociceptive action. In conclusion, MECN exerted the peripherally and centrally mediated antinociceptive activity via the modulation of the opioid/NO-mediated, but cGMP-independent, systems.