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Background: The composition and bioactivity of natural plant extract strongly depend on the extraction technique employed. Clinacanthus nutans Lindau (C. nutans) is a well-known medicinal plant in South-East Asia that has been traditionally used for the treatment of various diseases. Several conventional methods have been using for the extraction of bioactive compounds from C. nutans. However, extraction of fatty acids using supercritical carbon dioxide was not reported yet from this medicinal herbs. Objective: The main objective of the study is to examine the potential of supercritical carbon dioxide (scCO2) extraction of fatty acids from leaves and stems of C. nutans. Method: Fatty acid compositions were determined from leaves and stems of C. nutans oil extracted by scCO2 (temperature 45-65 °C, pressure 25-35 MPa) and compared to the results of Soxhlet extraction. Results: Supercritical CO2 extraction shows the highest oil recovery in both leaves (3.7%) and stems (1.6%) at pressure 35 MPa, temperature 65°C and 2 ml/min flow rate, which was closer to the yield of Soxhlet. The scCO2 yields presented a higher percentage of polyunsaturated fatty acids (PUFA), especially linoleic acid (C18:2n-6). Palmitic acid ranging from 42%- 47% in leaves and stems of C. nutans was found dominant saturated fatty acids (SFA) in both scCO2 and Soxhlet method. Conclusion: The current results indicate that leaves and stems of C. nutans could be a potential source of fatty acids, especially biologically active compounds.

Background: The preparations of commercially available Clinacanthus nutans (Burm.F) Lindau (Acanthaceae) (CN) leaf are gaining attention as an alternative cancer treatment, particularly in South East Asia. Multiple studies have suggested that CN has potential anticancer activities; however, the mechanism of these activities has remained elusive. Objective: This study evaluated the cytotoxic mechanisms of CN extracts in cancer cells. Methods: CN extracts were prepared from either fresh or dried leaves, using different solvents. Cytotoxicity of CN extracts was tested on the A549 (lung cancer), HeLa (cervical cancer), MCF-7 (breast cancer) and MDA-MB-231 (breast cancer) cell lines using the MTT assay. Flow cytometry was used to assess changes in the cell cycle profile, while Western blotting was used to examine microtubule stability. Finally, the mode of cell death was investigated using the Annexin V-FITC Apoptosis Detection Kit. Results: Aqueous Fresh (AQF) extract was prepared to simulate the ethnomedicinal use of CN, and reduce cell viability of MCF-7 cells with IC50 = 1.71 mg/mL. Some CN extracts have the ability to inhibit the proliferation of four different cancer cell lines after a 24 hour treatment. Annexin V assay results showed that acetone extracts of CN induced increments in the percentage of apoptotic cell death. However, flow cytometry results showed that the cancer cell cycle profile was not affected. Similarly, immunoblotting results also indicate that microtubule dynamics in MCF-7 cells were not altered. However, the aqueous extract, prepared to simulate the current ethnomedicinal use of CN leaves in cancer treatment, did not significantly inhibit cancer cell proliferation with an IC50 = 1.71 mg/mL. Conclusions: This study was the first to show that microtubules in cancer cells remain dynamic after treatment with CN extracts, effectively ruling interference of microtubule dynamics as the mode of cell death. AMD extract showed the highest effects of MCF-7 cell proliferation.

The development of new and effective pain medications that are natural is expected to have the benefits of treatment. The purpose of this study was to determine the antinociceptive effectiveness of a combination of 75% ethanol extract from dandang gendis leaves and 96% ethanol fraction of bakung leaf. In this study was used chemical method induction (Sigmund's method) in 25 male mice which were divided into 5 treatment groups, namely treatment group I as negative control was given CMC 0.5%, treatment group II as positive control was given acetosal 100 mg / kg BW, treatment groups III, IV and V were given combinations extract. Each group was given treatment orally, thirty minutes then induced with 0.6% acetic acid intraperitonially, after which it was observed and counted for the amount of stretching for sixty minutes. Data were analyzed by one-way ANOVA test. The results of this study showed that all treatment groups had a significant difference (p

Background: There has been an increasing trend in Clinacanthus nutans’ research, an important medicinal herb of Malaysia and Thailand, well known as an anti-viral, anti-cancer, and insect bite treatment. This study examines the trend in Clinacanthus nutans’ research from 2000 to 2019 and compares the contribution of research on this topic from different institutions and authors. Methods: Publications from the Scopus database were retrieved using keywords and identify top ten institutions/universities, list of prominent authors, top ten journals that published research, top five influential articles, top fifty cited papers, and global distribution of publications on Clinacanthus nutans. Microsoft Excel 2016, Wordcloud, SPSS version 26, and GunnMap 2 were used to analyse indicators. A total of 167 articles were identified from the Scopus database. All research publications were screened initially. Five articles (n=5) were removed due to the unavailability of the full-text version of the article. 162 articles were included in the final study. Results: Universiti Putra Malaysia and the Journal of Ethnopharmacology published the highest number of articles on Clinacanthus nutans. Herpes, antioxidant, phenolic, flavonoids, cancer, antimicrobial were common keywords identified using a word cloud. Over the past 20 years, the literature on Clinacanthus nutans has continuously grown, with the rate increasing after 2012. Conclusion: The prominent research on Clinacanthus nutans was based upon their identified and isolated bioactive constituents, and there is a need for more research on clinical trials.

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

Acanthaceae is popularly known as acanthus family which belong to mint order – lamiales. They are distributed from tropics to a temperate region such as India, Malaysia, Brunei, Indonesia, Brazil, Central America and Africa. Most members of this family are therapeutically important since they are in the up to date usage by ethnic communities. Andrographis paniculata, Clinacanthus nutans, Graptophyllum pictum, Hemigraphis alternata, Justicia gendarussa and Strobilanthes Crispus are some of the medicinal plants of Acanthaceae family. These plants are recognized for their biopharmaceutical potential usage in traditional medicine. These plants have a plethora of phytochemical compounds such as flavonoids, phenolic compounds, glycosides, terpenoids, benzenoids, quinine, triterpenoids and naphthoquinone present in various parts of the plant that plays a vital role in drug industries. The pharmacological properties of these plants such as anti-bacterial, anti-diabetic, anti-cancer, anti-oxidant, anti-inflammatory, anti-arthritis, hepatoprotective, anti-viral and anti-hypertensive are in general practice as an alternative and complementary medicine in both ethnobotanical and pharmacological fields. This article encompasses not only the comprehensive survey based on the electronic resources, scientific journals but also the books that summarize the botanical, phytochemical properties of these plants and also accentuate their significant role in both ethnobotanical and pharmacological fields. It is felt that this article would provide more insight into the health benefits of some plants of the Acanthaceae family.

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

Clinacanthus nutans was found to possess anti-venom, anti-inflammatory, analgesic, anti- diabetic, anti-rheumatism, antiviral and antioxidant properties. Silver nanoparticles are nanoparticles between 1nm to 100nm in size and play significant role in medicinal fields. Silver nanoparticles exhibit unique properties, such as excellent conductivity, chemical stability, catalytic and antimicrobial activity. Streptococcus mutans is usually discovered in human oral cavity and the main aetiological of tooth decay. There is no study on antibacterial effect of silver nanoparticles Clinacanthus nutans (AgNP-CN) against Streptococcus mutans reported to date. Therefore, objective of this study is to investigate antibacterial properties of silver nanoparticles Clinacanthus nutans against Streptococcus mutans. Streptococcus mutans was subcultured in brain heart infusion (BHI) broth and agar. AgNP-CN with different concentrations was tested against Streptococcus mutans via disc diffusion assay, minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC). Amoxicillin was used as positive control while DMSO and blank silver nanoparticles were used as negative control. Disc diffusion assay shows AgNP-CN inhibit Streptococcus mutans growth. AgNP-CN shows the greatest inhibition properties (15.3±1.53 mm) in comparison to Clinacanthus nutans leaves extract (6.0±0.01 mm) and blank silver nanoparticles (6.0±0.01 mm). MIC value for AgNP-CN is 2.5±0.01 mg/mL while amoxicillin is 0.007±0.01 mg/mL. Whereas MBC values for AgNP-CN is 2.5 mg/mL. Results are concentration dependent, with higher concentration shows better inhibition property. It can be concluded AgNP-CN possesses bactericidal properties against Streptococcus mutans.

Nowadays, herbs have become a popular form of healthcare. Clinacanthus nutans (Burm.f.) Lindau (family Acanthaceae) is popular in Asia, especially in the tropical countries. Extracts of C. nutans leaves are widely used to treat diseases of skin, anti-inflammatory, anti-virus, anticancer, antioxidant, antidiabetic, immunomodulatory, wound healing and analgesic activities, etc. This study determined the parameters of ethanol extract of C. nutans leaves including pH, total weight loss due to water, total ash content, heavy metal content, microbiological limits, pesticide residual content, presence and contents of C-flavones. The C-flavones such as shaftoside, orientin, isovitexin and vitexin have been found to be major flavonoids in the leaves of this plant. Therefore, we had used a two-step method using thin layer chromatography (TLC) and high pressure liquid chromatography (HPLC) for the rapid identification and quantification of the flavones C-glycosides in ethanol extract C. nutans leaves. These results provided useful information for the evaluation of quality of C. nutans raw materials and its commercial products. Moreover, that contributed to the establishment of extract standard from the leaves of C. nutans. The parameters of the ethanol extract of C. nutans leaves were obtained as follows: pH (5,23 ± 0,35), total weight loss due to water (15,62 ± 0,25%), total ash (13,85 ± 0,98%), microbiological limits are achieved the requirement of medicinal standards, content heavy metal and residue of pesticide are not detected. Contents of C-flavones: shaftoside (0,69 ± 0,04 mg/g), orientin (0,24 ± 0,008 mg/g), isovitexin (0,015 ± 0,003 mg/g), vitexin were very low or undetectable.

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 leaves is a medicinal plant with promising therapeutic effect. The objective of this study was to produce C. nutans powder using a spray dryer and to evaluate its physicochemical properties, followed by physical observation after 3 months of storage at room temperature (21±1°C). C. nutans juice with and without the addition of 5% (w/v) of modified corn starch (MCS) was spray dried at a feed flow rate of 23 rpm, inlet air temperature of 160°C and outlet air temperature of 100°C. Results showed that significantly lower (p

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.

The metabolomics approach successfully explained the possible neuroprotective effect of Clinacanthus nutans (Burm. f.) Lindau (CN) leaf extracts. Forty-four metabolites were putatively identified via proton Nuclear Magnetic Resonance (1H NMR and J-resolved NMR) metabolic profiling of CN leaf extracts in three types of solvents, namely water, 50% ethanol, and ethanol. Metabolite fingerprinting has efficaciously differentiated aqueous between the other two extracts. The variable importance of projection (VIP) showed that 30 metabolites were responsible for the discrimination of the extracts by component 1 in the Partial Least Square (PLS) score plot. The lipopolysaccharides (LPS)-induced murine microglial of the BV2 cell line successfully exhibited aqueous CN as the closest extract related to the nitrite oxide (NO) inhibitory activity via PLS biplot, with an IC50 value of 336.2 ± 4.7 µg/mL through Griess assay. The cytotoxicity assay also indicated that all CN extracts were non-toxic. Schaftoside, acetate, propionate, alanine, and clinacoside C were identified as the most potential biomarkers in the anti-inflammatory assay. Hence, the aqueous CN extract could be further investigated, particularly relating to the anti-neuroinflammation study.

Clinacanthus nutans Lindau (C. nutans) is a popular medicinal herb plant in Asia regions such as Malaysia, Indonesia, Thailand and China for its high polyphenols content. Various conventional extra...

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.

Isolation of anti-proliferative compounds from plants is always hindered by the complexities of the plant’s nature and tedious processes. Clinacanthus nutans (Burm. f.) Lindau is a medicinal plant with reported anti-proliferative activities. Our study aimed to isolate potential anti-proliferative compounds present in C. nutans plant. To start with, for our study, we came up with a strategy by first profiling the volatile compounds present in the leaf, stem and root of C. nutans using GC-MS. Comparing the plant’s volatile profiles greatly narrowed down our target of study. We decided to start with the isolation and characterization of a pentacyclic terpenoid, i.e., lupeol from the roots of C. nutans, as this compound was found to present abundantly in the roots compared to the leaf or stem. We developed a simple maceration and re-crystallization method, without the necessity to go through the fractionation or column chromatography for the isolation of lupeol. Characterizations of the isolated compound identified the compound as lupeol. The anti-proliferative activity of the isolated lupeol was further investigated against the MCF-7 cell line, which showed comparable anti-proliferative activity with the authentic lupeol and camptothecin. Our strategy to profile every part of the plant first, followed by selection of the most suitable plant part and targeted compound proved useful for further isolation and characterization bioactive compound from C. nutans.

The lack of specific treatment for chikungunya fever makes the need for anti-chikungunya virus agents more crucial. This study was conducted to evaluate 132 extracts obtained by sequential solvent extraction from 21 medicinal plants for cytopathic effect inhibitory activity using virus-infected Vero cells in two different sample introduction modes. Among the extracts, 42 extracts (31.8%) from 12 plants in the concurrent mode and three extracts (2.3%) from a plant in the non-concurrent mode displayed strong cytopathic effect inhibitory activity (cell viability ≥70%). Viral load quantification analysis unveiled that the extracts of Clinacanthus nutans (chloroform, ethyl acetate, and ethanol), Hydrocotyle sibthorpioides (ethanol), and Ocimum americanum (ethanol and methanol) hindered the release of viral progeny from the infected cells while the extracts of Ficus deltoidea (ethanol), Gynura bicolor (water), H. sibthorpioides (water), and O. americanum (chloroform and ethyl acetate) blocked the entry of virus into the cells. The extracts of Diodella sarmentosa (ethyl acetate), Diplazium esculentum (chloroform, ethyl acetate, and ethanol), and G. bicolor (ethanol) possessed virucidal effect and caused 5.41-log to 6.63-log reductions of viral load compared to the virus control. The results indicate that these medicinal plants are potential sources of anti-chikungunya virus agents that have varied modes of action.

Clinacanthus nutans is a well-known source for pharmaceutical drugs with most of the active ingredients classified as secondary metabolites. The study objectives are to explore the effect addition of different plant hormones alone or by combinations in the MS plant growth medium and factor of light sources towards induction of callus via in vitro culture. In this study, nodal explant of C. nutans was cultured on Murashige and Skoog Medium (MS) supplemented with different concentrations of Naphthaleneacetic acid (NAA) (0.5, 1.0, 1.5, and 2.0 mg/L) and 2,4-Dichlorophenoxyacetic acid (2,4-D) (0, 1, 2, 3 and 4 mg/L) at dark and light conditions. In hormone combination treatments, study was conducted with addition on same concentration of NAA and BAP in MS medium. Study with single hormone in MS medium resulted that only NAA had successfully induced callus at both light and dark conditions. Highest friable callus produced is at MS + 1.5 mg/L NAA in dark as 100% callus induced with 1.470 ± 0.225 g fresh weight. Next, study on the effect of NAA + BAP combinations in MS medium resulted that the treatment induced compact callus formation in both light and dark condition. The highest number of callus produced is in treatment of MS + NAA (0.1 mg/L) + BAP (0.1 mg/L) with 0.569 ± 0.159 callus fresh weight. To conclude single treatment callus 1.5 mg/L NAA alone at dark condition was best to produce friable and proliferative callus.

The use of metabolomics as a comprehensive tool in the analysis of metabolic profiles in disease progression and therapeutic intervention is rapidly advancing. Yet, a single analytical platform could not be applied to cover the entire spectrum of a biological sample’s metabolome. In the present paper, multi-platform metabolomics approaches were explored to determine the diverse rat sera metabolites extracted from intracerebroventricular lipopolysaccharides (LPS)-induced neuroinflammed rats treated with oral therapeutic interventions of positive drug (dextromethorphan, 5 mg/kg BW); with Clinacanthus nutans (CN) aqueous extract (CNE, 500 mg/kg BW); and with phosphate buffer saline (PBS) as the control group for 14 days. Analyzed by nuclear magnetic resonance (NMR) and liquid chromatography-mass spectrometry (LC-MS) techniques, this study depicted the potential of metabolites associated with neuroinflammation and verified by MetDisease. The key observations in the perturbed metabolic pathways that showed ameliorative effects were linked to the class of amino acid and peptide metabolism involving valine, leucine, and isoleucine biosynthesis; phenylalanine, tyrosine, and tryptophan biosynthesis; and phenylalanine metabolism. Lipid metabolism of arachidonic acid metabolism, glycerophospholipid metabolism, terpenoid backbone biosynthesis, and glycosphingolipid metabolism were also affected. Current findings suggested that the putative biomarkers, especially lysophosphatidic acid (LPA) and 5-diphosphomevalonic acid from glycerophospholipid and squalene/terpenoid and cholesterol biosynthesis, respectively, showed the ameliorative effects of the drug and CN treatments by controlling cell differentiation and proliferation. Our study proved that the complex and dynamic sera profiling affected during the CN treatment was greatly influenced by the analytical platform selection as integration between the two data yielded a more holistic summary of the metabolite pattern changes. Hence, an evidence-based herb, such as CN, can be used for novel diagnostic tools in the quest for ethnopharmacological studies.

Cervical cancer is a leading cause of death in women, worldwide, and second in Malaysia. Nowadays, modern cancer treatment comes with negative and adverse side effects to the patients. Traditional medicinal plants such as Clinacanthus nutans has been locally recognized for its medicinal properties and was claimed for cancer treatment. According to our pilot study, a fraction of C. nutans has shown a potent inhibition on human cervical cancer cells, SiHa, in vitro. Hence, this study aimed to investigate the underlying anticancer mechanism of a semi-purified fraction (SF1) of C. nutans towards cervical cancer cells. Bioassay guided fractionation and further purification was carried out. All collected fractions of C. nutans were examined for cytotoxicity activity towards HeLa, SiHa and NIH cells by MTT Assay. SF1 exhibited better cytotoxic activity with best growth inhibition against SiHa cells (IC50 value = 9.60 μg/mL). Cytoselective effects were shown as no cytotoxicity observed in the treated normal NIH cells. SF1 induced early apoptosis event in SiHa cells as resulted by FITC-Annexin V/propidium iodide staining by flowcytometric analysis. The protein expression by flowcytometry using specific antibody conjugated fluorescent dye showed up-regulation of Bax and down-regulation Bcl-2 as well as increment of cytochrome C levels upon treatment with SF1. In conclusion, the findings suggested that SF1 demonstrated antiproliferative effects and induced apoptosis in SiHa cells through up-regulation of Bax and down-regulation of Bcl-2 via mitochondrial-dependent pathway.

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 emergence of new diseases and the increased use of antibiotics have led to the development of resistant bacterial strains. Thus, there is greater attention to seek new antibacterial agents from the natural sources for combating fish diseases in the aquaculture industry. The present study evaluated the bactericidal efficacy of crude methanolic and aqueous extracts from Polygonum chinense, Syzygium polyanthum, Premna foetida, Pimenta dioica, Brucea javanica, Vitex negundo, Alpinia conchigera and Clinacanthus nutans against Vibrio harveyi, Vibrio alginolyticus, Vibrio parahaemolyticus and Aeromonas hydrophila using disc diffusion method. The results showed that methanolic extracts of P. dioica, P. foetida and P. chinense, and aqueous extracts of P. dioica and S. polyanthum showed moderate to strong activity (10.8 to 17.2 mm) against all the tested bacteria. These five potential crude extracts were fractionated using liquid-liquid extraction method to obtain the methanol, dichloromethane and ethyl acetate fractions. Among the fractions, ethyl acetate fraction showed the highest activity against all tested bacteria, with minimum inhibition concentration (MIC) values between 0.625 and 10.000 mg/mL. In addition, the five potential crude extracts had low to moderate toxicity with LC50 >100 μg/mL using brine shrimp cytotoxicity assays. The results of this study indicated that methanolic extracts of P. chinense and P. foetida that showed high bactericidal activity and low toxicity could be good potentials for use in fish culture.

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.

Ethnopharmacological relevance The extensive biodiversity of plants in Southeast Asia and inadequate research hitherto warrant a continued investigation into medicinal plants. On the basis of a careful review of fresh medicinal plant usage to treat cancer from previous ethnobotanical interviews in Singapore and from the traditional uses of the indigenous plants, fresh leaves of seven locally grown medicinal plant species were evaluated for anti-proliferative activity. Aim of the study To evaluate the anti-proliferative activity of local medicinal plant species Clausena lansium Skeels, Clinacanthus nutans (Burm. f.) Lindau, Leea indica (Burm. f.) Merr., Pereskia bleo (Kunth) DC., Strobilanthes crispus (L.) Blume, Vernonia amygdalina Delile and Vitex trifolia L. Materials and method Fresh, healthy and mature leaves of the seven medicinal plants were harvested from various locations in Singapore and Malaysia for Soxhlet, ultrasonication and maceration extractions in three different solvents (water, ethanol and methanol). Cell proliferation assay using water soluble tetrazolium salt (WST-1) assay was performed on twelve human cancer cell lines derived from breast (MDA-MB-231, T47D), cervical (C33A), colon (HCT116), leukemia (U937), liver (HepG2, SNU-182, SNU-449), ovarian (OVCAR-5, PA-1, SK-OV-3) and uterine (MES-SA/DX5) cancer. Results A total of 37 fresh leaf extracts from seven medicinal plants were evaluated for their anti-tumour activities in twelve human cancer cell lines. Of these, the extracts of C. lansium, L. indica, P. bleo, S. crispus, V. amygdalina and V. trifolia exhibited promising anti-proliferative activity against multiple cancer cell lines. Further investigation of selected promising leaf extracts indicated that maceration methanolic extract of L. indica was most effective overall against majority of the cancer cell lines, with best IC50 values of 31.5 ± 11.4 µg/mL, 37.5 ± 0.7 µg/mL and 43.0 ± 6.2 µg/mL in cervical C33A, liver SNU-449, and ovarian PA-1 cancer cell lines, respectively. Conclusion The results of this study provide new scientific evidence for the traditional use of local medicinal plant species C. lansium, L . indica, P. bleo, S. crispus, V. amygdalina and V. trifolia in cancer treatment. These results highlight the importance of the upkeep of these indigenous plants in modern society and their relevance as resources for drug discovery.

The present study used in vitro and in silico techniques, as well as the metabolomics approach to characterise α-glucosidase inhibitors from different fractions of Clinacanthus nutans. C. nutans is a medicinal plant belonging to the Acanthaceae family, and is traditionally used to treat diabetes in Malaysia. n-Hexane, n-hexane: ethyl acetate (1:1, v/v), ethyl acetate, ethyl acetate: methanol (1:1, v/v), and methanol fractions were obtained via partitioning of the 80% methanolic crude extract. The in vitro α-glucosidase inhibitory activity was analyzed using all the fractions collected, followed by profiling of the metabolites using liquid chromatography combined with mass spectrometry. The partial least square (PLS) statistical model was developed using the SIMCA P+14.0 software and the following four inhibitors were obtained: (1) 4,6,8-Megastigmatrien-3-one; (2) N-Isobutyl-2-nonen-6,8-diynamide; (3) 1′,2′-bis(acetyloxy)-3′,4′-didehydro-2′-hydro-β, ψ-carotene; and (4) 22-acetate-3-hydroxy-21-(6-methyl-2,4-octadienoate)-olean-12-en-28-oic acid. The in silico study performed via molecular docking with the crystal structure of yeast isomaltase (PDB code: 3A4A) involved a hydrogen bond and some hydrophobic interactions between the inhibitors and protein. The residues that interacted include ASN259, HID295, LYS156, ARG335, and GLY209 with a hydrogen bond, while TRP15, TYR158, VAL232, HIE280, ALA292, PRO312, LEU313, VAL313, PHE314, ARG315, TYR316, VAL319, and TRP343 with other forms of bonding., Graphical abstract fx1, Highlights • The antidiabetic potential of various extracts of C. nutans leaf has been evaluated. • Multivariate data analysis identified 4 α-glucosidase inhibiting compounds. • In silico study predicted the protein-inhibitors interaction.

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

Objective: This study was conducted to investigate the antiproliferative activity of extracts of Clinacanthus nutans leaves against human cervical cancer (HeLa) cells. Methods: C. nutans leaves were subjected to extraction using 80% methanol or water. The methanol extract was further extracted to obtain hexane, dichloromethane (DCM), and aqueous fractions. The antiproliferative activity of the extracts against HeLa cells was determined. The most cytotoxic extract was furthered analyzed by apoptosis and cell cycle assays, and the phytochemical constituents were screened by gas chromatography-mass spectrometry (GC-MS). Results: All of the extracts were antiproliferative against HeLa cells, and the DCM fraction had the lowest IC50 value of 70 μg/mL at 48 h. Microscopic studies showed that HeLa cells exposed to the DCM fraction exhibited marked morphological features of apoptosis. The flow cytometry study also confirmed that the DCM fraction induced apoptosis in HeLa cells, with cell cycle arrest at the S phase. GC-MS analysis revealed the presence of at least 28 compounds in the DCM fraction, most of which were fatty acids. Conclusion: The DCM fraction obtained using the extraction method described herein had a lower IC50 value than those reported in previous studies that characterized the anticancer activity of C. nutans against HeLa cells.

The analytical method used in the quality control of Clinacanthus nutans leaves has not been well developed. Therefore, this study aimed to develop a simple analytical method to predict α-glucosidase inhibitory activity of this herb based on its infrared fingerprinting. The dried extracts obtained from maceration using solvents with different polarities were evaluated for the α-glucosidase inhibitory activity and analysed through infrared spectroscopy. Multivariate data analysis was performed by correlating the bioactivity and infrared spectrum of each extract using partial least square method. The loading plot from multivariate data analysis revealed that C H and C O infrared signals from terpenoids in the extract were positively correlated with the α-glucosidase inhibitory activity. The developed partial least square model was validated through a testing on the external samples. The result concludes that the developed model is valid and capable of predicting α-glucosidase inhibitory activity of the external samples.

Clinacanthus nutans, an herbal shrub belonging to the Acanthaceae family, is traditionally used as a functional food to treat various ailments in Malaysia and Indonesia. Although the polar fraction of this plant shows non-toxic effect, the toxicity of the non-polar extract is not reported so far. The present study aimed to assess the toxic effect and determine the lethal concentration of this non-polar fraction using zebrafish embryos. The n-hexane fraction was partitioned from the crude extract of C. nutans obtained using 80% methanolic solution. After spawning of the adult male and female zebrafish, the eggs were collected, transferred into a 96-well plate and incubated with the n-hexane fraction at concentrations of 15.63 μg/ml, 31.25 μg/ml, 62.5 μg/ml, 125 μg/ml, 250 μg/ml and 500 μg/ml in 2% DMSO. The survival and sublethal endpoint were assessed, the mortality and hatchability rates were calculated based on microscopic observation, while the heartbeat rate was measured using DanioScope software. The median lethal concentration (LC50) of the C. nutans n-hexane fraction, which was determined using probit analysis, was calculated to be 75.49 μg/mL, which is harmful. Moreover, gas chromatography-mass spectrometry (GC–MS) analysis revealed the presence of palmitic acid, phytol, hexadecanoic acid, 1-monopalmitin, stigmast-5-ene, pentadecanoic acid, heptadecanoic acid, 1-linolenoylglycerol and stigmasterol in the n-hexane fraction. Keywords: Clinacanthus nutans, Danio rerio, DanioScope, Median lethal concentration, Teratogenicity, Probit analysis

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.

Surface sterilization is a vital step in preparation of healthy and viable explants in tissue culture. Most surface contaminants can be eliminated by surface sterilization with a suitable sterilizing agent. The study aimed to present an effective disinfection method for Clinacanthus nutans shoot regeneration using nodal segments. A total of four different sterilization approaches were conducted by treating nodal explants with various concentrations of sterilizing agent. Sterilizing agents used were Rhizophora apiculata Pyroligneous acid (PA), sodium hypochlorite (Clorox) thiophanate-methyl (fungicide), and Mercuric chloride (HgCl2). Nodal explant then was cultured on plant growth regulator-free Murashige and Skoog (MS) basal medium. This study sterilizing agents revealed that PA showed strong bactericidal activity. However, it led to a high number of fungal contaminations. The pyroligneous acid did not exhibit a strong potential as a disinfectant for C. nutans nodal explant. Overall, HgCl2 exhibits the best reduction in fungal contamination and gives a significant result with thiophanate-methyl fungicide. Surface sterilization with mercuric chloride (0.2%) for 1 hour was the optimum concentration and duration, which resulted in the highest percentage of nodal explant survival and viability. All viable nodal segments developed into shoots. It had been concluded that the best surface sterilization agent was HgCl2.

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.