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Ethnopharmacological Relevance: Indigofera linifolia (L.f.) Retz. is used in subcontinent for liver disorders, in wounds, febrile eruption and as diuretic., Aim of Study: The current study evaluates the protective effects of the methanol extract of Indigofera linifolia (ILM) on CCl 4 -induced endoplasmic reticulum (ER) stress in liver of rat., Methods: ILM was analyzed for phytochemical classes, total phenolic (TPC) and flavonoid content (TFC) as well as multidimensional in vitro antioxidant assays. Male (Sprague Dawley) rats were dispersed into seven groups (6 rats/group) receiving 0.9% saline (1 ml/kg bw), CCl 4 (1 ml/kg bw) diluted in olive oil (3:7 v/v), silymarin (200 mg/kg bw) + CCl 4 (30% v/v), ILM (150 mg/kg bw) + CCl 4 (30% v/v), ILM (300 mg/kg bw) + CCl 4 and ILM alone (either 150 mg/kg bw or 300 mg/kg bw)., Results: ILM extract was constituted of different phytochemical classes. Co-administration of ILM along with CCl 4 to rat revert the level of alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP) and total bilirubin in blood serum and antioxidant parameters in liver. Further, CCl 4 increased the level of ER stress markers and inflammatory mediators while decreased level of GCLC and Nrf-2 in liver tissues of rat. CCl 4 -induced histopathological variations were reduced with ILM co-administration in liver tissues., Conclusion: The results suggest that active phyto-constituents of I. linifolia might be responsible for its antioxidant, anti-inflammatory and gene-regulating activities., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Ethnopharmacological Relevance: Aerial parts of Vincetoxicum arnottianum (Wight) Wight (Family Apocynaceae) are used by local communities for inflammation, healing of wound and injuries and also for urticaria., Aim of Study: Extract/fractions of V. arnottianum were evaluated for potential anti-inflammatory activity in rat., Methods: Methanol extract of aerial parts of V. arnottianum (VAM) was partitioned on polarity for n-hexane (VAH), ethyl acetate (VAE), butanol (VAB) and aqueous (VAA) fractions. The extract/fractions were evaluated during in vitro assay for protection against heat induced protein denaturation and Carrageenan induced paw inflammation in rat. VAM and VAE were evaluated for anti-inflammatory potential against formalin and Freund's complete adjuvant (FCA) induced inflammation in paw of rat while croton oil induced inflammation in ear of rat, respectively. The level of inflammatory mediators; IL-17, IL-1β, IL-6, TNF-α and nitric oxide (NO) was estimated in serum of rat., Results: All the extract/fractions used in this study exhibited anti-inflammatory activity. However, VAE (300 mg/kg) exhibited potential anti-inflammatory activity in carrageenan (78.06 ± 4.6%), formalin (54.71 ± 0.34%) and croton oil (73.12 ± 1.9%) induced edema in rat. In FCA induced inflammation model VAM and VAE showed admiring proficiencies against alteration of body weight and organ weight indices, paw edema and histological studies. In serum increased level of pro-inflammatory cytokines (IL-1β, TNF-α, IL-6, IL-17) and NO during adjuvant-induced inflammation were more efficiently restored with VAE treatment to rat. Presence of polyphenolics; rutin, gallic acid, caffeic acid, apigenin, myricetin and quercetin was indicated in VAE., Conclusion: The results suggest the presence of anti-inflammatory constituents in V. arnottianum., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Ethnopharmacological Relevance: Leaves of Alnus nitida are used by local communities for the management of diabetes and in inflammatory disorders., Methods: Powder of shade dried leaves of A. nitida was extracted with methanol (ANME) and fractionated in escalating polarity i.e n-hexane (ANHE), chloroform (ANCE), ethyl acetate (ANEE) and soluble residual aqueous fraction (ANAE). The extract/fractions were evaluated for antidiabetic in vitro assays; α-amylase, α-glucosidase and dipeptidyl peptidase-4 (DPP-4). The in vivo investigations were carried out on ANEE and ANAE (100 mg/kg; 200 mg/kg, p.o.) in alloxan (125 mg/kg i.p.) induced hyperglycemic rats. Serum analysis was performed on liver, pancreas and kidney function markers. Analysis of antioxidant enzymes and genotoxic studies were carried out on pancreas, liver and kidneys tissues. GC-MS analysis was performed on ANME whereas HPLC analysis was carried out on ANME, ANEE and ANAE., Results: Preliminary in vitro assays indicated appreciable antidiabetic activity of ANEE and ANAE against α-amylase, α-glucosidase and DPP-4 assay. Furthermore, in vivo antidiabetic effect of ANEE and ANAE was inveterate by anti-hyperglycemic action in normal glucose loaded and diabetic glucose loaded animals. Single dose of alloxan (125 mg/kg) decreased the level of insulin and high density lipoprotein while raised the level of amylase and lipase, ALT, AST, total lipids, triglycerides, cholesterol, creatinine, BUN, CPK, CK-Mb in serum. Concentration of H 2 O 2 , lipid peroxidation (TBARS) and nitrite was increased (P < 0.05) whereas level of tissue protein, glutathione content (GSH) and antioxidant enzymes decreased in pancreas, liver and kidneys as compared to control group. Administration of ANEE and ANAE for 14 days after induction of diabetes decreased the hyperglycemia and restored the level of these parameters. Histopathological and genotoxic studies also endorsed the defensive strategies of ANEE and ANAE. GC-MS analysis of ANME demonstrated the presence of antidiabetic constituents i.e. linalool, Vitamin E and phytol., Conclusion: Results obtained in this study suggests antidiabetic and antioxidant abilities and provides the scientific proof of the folklore medicine., Competing Interests: Declaration of competing interest Authors declare no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Ethnopharmacological Relevance: Abutilon pannosum is used in Pakistan for bladder inflammation, diuretic, lung disorders, diabetes and in lowering pyrexia., Methods: Amount of total phenolic content, total flavonoid content and HPLC analysis of APM for the presence of polyphenolics were carried out. Antioxidant activity was determined by using different in vitro antioxidant assays. Amelioration effects of APM (200 mg/kg body weight and 400 mg/kg body weight) against CCl 4 induced kidney and lung toxicity in rat was assessed by determining level of antioxidant enzymes, lipid peroxidation products, cytokines (TNF-α, IL-1β and IL-2), comet assay and histological analysis., Results: HPLC-DAD studies of APM indicated the presence of rutin (0.635 ± 0.011 μg/mg dry extract), gallic acid (1.07 ± 0.043 μg/mg dry extract) and catechin (0.246 ± 0.08 μg/mg dry extract) and considerable quantity of total phenolic (55.485 ± 0.85 mg GAE/g dry extract) and total flavonoid content (19.90 ± 0.58 mg Rutin/g dry extract). During in vitro antioxidant assays APM showed significant (p < 0.05) activity for iron chelation, reducing potential, inhibition of β-carotene oxidation while moderate potential for scavenging of DPPH, hydroxyl, nitrite and phosphomolybdenum radical. Administration of CCl 4 to rat severely depleted the activity level of catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD), and reduced glutathione (GSH) concentration while appreciably increased the concentration of thiobarbituric acid reactive substances (TBARS), H 2 O 2 , nitrite, TNF-α, IL-1β and IL-2 in lung and kidney tissues of rat. Comet length, % DNA in tail and tail moment significantly (p < 0.01) increased in lung and kidney cells of CCl 4 intoxicated rat. Further, injuries in lung and kidney tissues were recorded with CCl 4 induced toxicity in rat. The rats treated with APM along with CCl 4 exhibited an appreciable level of restoration of the altered parameters towards the control animals., Conclusion: The results of this study suggested the protective potential of APM in CCl 4 intoxicated rats for kidney and lung injuries., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Ethnopharmacological Relevance: Fagonia olivieri (Zygophyllaceae) is used in Pakistan for management of diabetes and for treatment of stress related disorders of liver and kidneys. The objective of present study was to evaluate antidiabetic activity of F. olivieri using streptozotocin-nicotinamide induced diabetic rat model., Methods: Powder of the whole plant of F. olivieri was extracted with methanol (FOME) and the residue was fractionated with n-hexane (FOHE), chloroform (FOCE), ethyl acetate (FOCE), n-butanol (FOBE) and the remaining soluble aqueous fraction (FOAE). The extract/fractions were evaluated for in vitro inhibition of α-amylase and α-glucosidase and for the antidiabetic studies in Sprague-Dawley rats. FOAE was evaluated in 21 day chronic multiple dose study. Glibenclamide (10 mg/kg) and FOAE were used at a concentration of 200 mg/kg (164.8 μg of catechin) and 400 mg/kg (329.6 μg of catechin) in streptozotocin (STZ)-induced diabetic rats. Various serum parameters, blood cell parameters and oxidative stress induced parameters of the liver and kidneys were investigated., Results: Results showed that aqueous fraction of F. olivieri (FOAE) potentially inhibited activity of pancreatic α-amylase, intestinal maltase and sucrase. Administration of FOAE (200 mg/kg; 400 mg/kg) to diabetic rats for 21 days increased body weight, hemoglobin and its indices, RBCs, platelets, lymphocytes and neutrophils against diabetic group. In serum level of glucose, transaminases, ALP, LDH, total bilirubin, creatinine and lipids decreased while insulin, total protein and albumin increased in contrast to diabetic group. Further, level of cytokines; TGF-β1, TNF-α and IL-6 in serum and altered level of antioxidant enzymes, glutathione and lipid peroxides of liver and kidney tissues restored towards control animals., Conclusion: This study demonstrated that FOAE inhibited activity of digestive enzymes and exhibited antioxidant, anti-inflammatory and antihyperglycemic activity in STZ-nicotinamide prompted diabetic rats and suggested that FOAE might be used as a therapeutic agent for the management of type-2 diabetes., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Ethnopharmacological Relevance: The leaves of W. heynei (family: Rubiaceae) are used by the folklore in swelling, wounds and body aches. In this study anti-inflammatory potential of W. heynei leaves has been evaluated during in vitro studies and in rat., Methods: Methanol extract of the leaves of W. heynei (WHLM) and its fractions; n-hexane (WHLH), chloroform (WHLC), ethyl acetate (WHLE), n-butanol (WHLB) and residual soluble aqueous (WHLA) were screened for phytochemical analysis and several active constituents (alkaloids, flavonoids, saponins, tannins, terpenoids, β-carotene and lycopene) were also quantified. Heat induced albumin denaturation assay and in vitro cell cultures was carried out for in vitro anti-inflammatory activity, while various in vivo assays like TPA induced ear edema, croton oil induced anus edema, formalin and carrageenan-induced hind paw edema was investigated in Sprague-Dawley rats. Alterations on levels of tumor necrosis factor (TNF-α), Interleukin-1β (IL-1β), IL-6 and prostaglandins (PGE 2 ) induced with WHLE was studied in serum after carrageenan induced paw edema in rat. Meanwhile, the dose dependent WHLE inhibition of NFκB pathway via regulation of the phosphorylation of IKKs, IκBα, and p65 subunit was studied in LPS-induced rat peritoneal macrophages. On account of marked anti-inflammatory activity of WHLE its bioactive components were analyzed by HPLC-DAD analysis., Results: The phytochemical analysis yielded alkaloids, saponins, tannins, coumarins, glycosides, quinones and vitamin C in WHLM and in all fractions. Fraction (WHLE) was enriched with alkaloids (20.20 ± 2.5%), flavonoids (25.26 ± 2.11%) and tannins (307.2 ± 2.03 mg of GAE/g of extract), while terpenoids (21.60 ± 1.65%) were the major constituents of WHLH. Ethyl acetate fraction convincingly protected heat induced albumin denaturation. WHLE exhibited highest edema inhibition in models of TPA-induced ear edema (74.51 ± 2.05) and croton oil-induced anal edema (75.38 ± 2.83). The pretreatment with WHLE significantly (p < 0.05) reduced the paw edema with formalin (78.99 ± 2.26%) assessed after 6 h and in carrageenan (75.71 ± 4.46%) was detected after 4 h. Level of anti-inflammatory markers; IL-1β, IL-6, TNF-α and PGE 2 in carrageenan induced paw edema in serum of rat was significantly (p < 0.001) decreased with WHLE pretreatment to rat. WHLE significantly inhibited the NFκB by reducing the phosphorylation of IKKs, IκBα, and p65 subunit in LPS-induced inflammation in rat peritoneal macrophages. HPLC-DAD analysis of WHLE indicated the presence of rutin, gallic acid, catechin, caffeic acid and myricetin., Conclusions: It is concluded that WHLM fractions have marked anti-inflammatory activity and this study endorsed the folklore use of W. heynei leaves for swelling related disorders., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Ethnopharmacological Relevance: Oil extracted from Parrotiopsis jacquemontiana stem traditionally used for wound healing, body aches and dermatitis. In this study we have evaluated oil for its phytoconstituents, antioxidant, antimicrobial and wound healing activities., Methods: Phytochemical characterization of oil was determined by standard qualitative procedures, gas chromatography mass spectrometry technique (GC-MS) and Fourier transform infra-red spectroscopy (FT-IR). The in vitro antioxidant aptitude was determined by scavenging of DPPH radical, hydroxyl ion, nitric oxide, inhibition of β-carotene bleaching assay and iron chelation power assay. The antimicrobial potential of oil was investigated by disc diffusion method against multidrug resistant (MDR) bacterial isolates and fungal strains. Wound healing was performed in vivo with determination of wound contraction rates, histopathology, hemostatic potential and hydroxyproline estimation., Results: GC-MS analysis indicated that oil was constituted mainly of 2, 6-dimethyl-8-oxoocta-2, 6-dienoic acid, methyl ester (18.2%), syringol (17.8%), catechol (12.4%), guaiacol (5.2%), p-cresol (5.4%) and phenol, 2-propyl- (3.7%). FT-IR analysis revealed several important functional groups in its chemical composition especially phenolic O-H compound stretching. Scavenging of DPPH radical, hydroxyl ion, nitric oxide, inhibition of β-carotene oxidation and iron chelation power assays indicated strong antioxidant activities of oil. Further it efficiently inhibited growth of multidrug resistant isolates of Staphylococcus aureus, S. lugdenesis, Klebsiella pneumoniae, Escherichia coli, Coagulase -ve staphylococci and Pseudomonas aeruginosa. The minimum inhibitory concentrations ranged between (32-256) (μg/mL) of oil. The oil also strongly inhibited the growth of various fungal isolates with low level of minimum inhibitory concentrations (64-256) μg/mL. Remarkable rate for wound closure and epithelization, hemostatic potential and marked increase (p < 0.05) in hydroxyproline content was observed for oil during wound healing in rat., Conclusion: The results suggested that oil can be used as a potential source of wound healing therapeutics., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Ethnopharmacological Relevance: Stem bark of Alnus nitida (Spach) Endl. (family Betulaceae) is used by local communities in northern areas of Pakistan as a remedy for swelling, injuries and pain. However no pharmacological study of this plant has been reported to confirm these activities. In this study anti-inflammatory and analgesic effect of A. nitida stem bark have been evaluated., Methods: Powder of the stem bark of A. nitida was extracted with methanol (ANBM) and partitioned in escalating polarity to get the n-hexane (ANBH), chloroform (ANBC), ethyl acetate (ANBE) and the residual soluble aqueous (ANBA) fractions. The methanol extract and derived fractions were evaluated for anti-inflammatory activity by using in vitro heat induced albumin denaturation assay and various in vivo assays; carrageenan-induced hind paw edema method, Freunds' complete adjuvant induced arthritis, histamine induced paw edema and xylene induced ear edema in Sprague Dawley rat. The extracts/fractions were also evaluated for analgesic effects by using hot plate analgesic test and acetic acid induced writhing test in rat. The ANBM composition was analyzed by HPLC-DAD and GC-MS analysis., Results: Results of heat induced albumin denaturation activity indicated that among the extract/fractions ANBC at concentration range of 100-500µg/ml remarkably protected the heat induced albumin denaturation. The pretreatment with ANBC significantly reduced the carrageenan induced edema with 90.81±1.6% after 4h, comparing with 86.63±3.42% reduction produced by the reference drug diclofenac potassium. Histopathological alterations of the gastric and hind paw were decreased with the extract/fractions. Furthermore, anti-inflammatory effects of ANBC were evident in Freunds' complete adjuvant induced arthritis, histamine induced paw edema and xylene induced ear edema. The latency time in hot plate analgesic assay with ANBC (61.59±0.38%) after 90min was comparable to standard drug morphine (69.31±2.67%) and aspirin (67.24±2.08%). Similarly ANBC significantly (p<0.01) inhibited the pain sensation in acetic acid induced writhing test in rat. HPLC-DAD analysis of ANBM indicated the presence of gallic acid, catechin and rutin while the GC-MS analysis of ANBM indicated the presence of 30 compounds predominantly of neophytadiene, 3,7,11,15 tetramethyl-2-hexadecen-1-ol, phytol, vitamin E and linalool., Conclusion: The results of this study suggested that the presence of polyphenols, sterols, terpenoids and other constituents might contributed towards the anti-inflammatory and analgesic activities of the crude methanol extract of A. nitida bark and its derived fractions. This study endorsed the folklore use of A. nitida bark for inflammation related disorders., (Copyright © 2017 Elsevier Ireland Ltd. All rights reserved.)

Ethnopharmacological Relevance: Branched cancerwort, Kickxia ramosissima (Wall.) Janchen (Scrophulariaceae) is traditionally used for the treatment of inflammatory disorders such as rheumatism, diabetes, jaundice and for activation of immune system. Local communities also used this plant for the treatment of spleen enlargement, as febrifuge and in dysmenorrhea. In this investigation antipyretic, analgesic and anti-inflammatory effects of K. ramosissima have been evaluated., Methods: Dried powder of the whole plant of K. ramosissima was extracted with methanol (KRM) and partitioned with solvents to obtain the n-hexane (KRH), chloroform (KRC), ethyl acetate (KRE), n-butanol (KRB) and the residual aqueous (KRA) fraction. KRM and the derived fractions were analyzed for the phytochemical constituents, yeast induced pyrexia, analgesic and anti-inflammatory activities by using carrageenan and Freunds' complete adjuvant-induced paw edema model in rat. On account of appreciable effects of KRM in the aforesaid models, KRM was subjected to the carrageenan induced air pouch model in rat. The exudate of air pouch was analyzed for the count of neutrophils, monocytes, lymphocytes and WBCs and for the estimation of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), nitric oxide (NO) and prostaglandin (PGE2)., Results: Phytochemical investigation of KRM indicated the existence of tannins, flavonoids, alkaloids, coumarins, cardiac glycosides, saponins, terpenoids and phlobatannins. Maximum concentration of total phenolic was determined in KRB followed by KRM while reverse was true for total flavonoids contents. KRM (200mg/kg) distinctly decreased the rectal temperature in yeast induced pyrexia comparable to standard, paracetamol. Pain sensation was effectively inhibited at 200mg/kg p.o. of KRM and KRB as manifested by a decrease (P<0.001) in count of writhing induced with acetic acid and increase of latency time in hot plate. Anti-inflammatory effects of KRM were evident and edema formation induced with carrageenan and Freunds' complete adjuvant-induced paw edema in rat was significantly (P<0.001) inhibited. Count of neutrophils, monocytes, lymphocytes, and WBCs along with inflammatory mediators; IL-6, NO, TNF-α and PGE2 was significantly (P<0.001) decreased in subcutaneous air pouch exudate with KRM. HPLC analysis of KRM indicated the presence of gallic acid, caffeic acid, rutin, catechin and myricetin., Conclusion: The results suggested the anti-inflammatory effects of KRM in inflammation related disorders, might be attributed by the presence of active phytoconstituents; flavonoids, saponins and terpenoids., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)

Aim of the Study: Asthma is an ailment of airways characterized by activation of the T helper (Th) 2 lymphocytes and subsequent movement of inflammatory cells. Boerhavia procumbens of family Nyctaginaceae is locally used for the treatment of asthma, cough, hemorrhoids, dropsy, cardiac, eyes and kidney problems. We have evaluated its methanol extract (BPM) as a therapeutic candidate for asthma against toluene diisocyanate (TDI) allergic model in rat. The BPM extract was obtained from the whole plant of B. procumbens in methanol. Sprague-Dawley male 36 rats (200-250 g) were categorized into 6 groups having six rats in each category. The animals were provoked (10%) and sensitized (5%) by TDI. Animals of groups I-III were vehicle control (ethyl acetate), diseased control (TDI) and reference control (TDI+dexamethasone {2.5mg/kg bw}), respectively. Animals of group IV (TDI+200mg/kg bw) and group V (TDI+400mg/kg bw) were administered with BPM whereas group VI was administered with 400mg/kg bw alone of BPM. Protective effects of BPM were determined by counting the number of leucocytes and estimation of interleukines in blood, bronchoalveolar lavage (BAL) and in in vitro culture of spleen cells. Estimation of antioxidant enzymes, lipid peroxides and H2O2 and histopathology of lungs were carried out for antioxidant potential of plant extract used., Results: Methanol extract of B. procumbens suppressed the asthmatic symptoms and inhibited the infiltration of eosinophils and lymphocytes in lungs of TDI provoked rats. Administration of BPM to TDI provoked rats, dose dependently, inhibited the release of interleukins (IL)-2 in serum and IL-4, IL-6 interferon gamma (IFN-γ) in bronchoalveolar lavage (BAL) and in in vitro culture of spleen cells, and ameliorated the oxidative stress in lung tissues. Quantitative scoring of the lung histopathology exhibited protective effects of BPM and the inflammation, mucus, thickening of peribronchial smooth muscle layer and subepithelial deposition of collagen induced with TDI were ameliorated. The BPM has the anti-inflammatory properties that may be used to treat the asthma and inflammatory related ailments., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

Introduction: Carbon tetrachloride exerts its toxicity in rat kidneys through oxidative stress. Launaea procumbens is used ethnopharmacologically in renal disorders. Its methanolic, chloroform, ethyl acetate, and n-hexane fractions were evaluated against CCl(4)-induced nephrotoxicity in rat., Materials and Methods: Among 16 groups, one was given CCl(4) of 3ml/kg body weight (30% in olive oil i.p.) twice a week for 4 weeks. Others were orally fed with extracts (100, 200mg/kg b.w.) or rutin (50mg/kg b.w.) after 48h of CCl(4) or 200mg/kg b.w. of extracts without CCl(4)-treatment twice a week for 4 weeks., Results: CCl(4) exposure led to a significant oxidative stress in kidneys which was remarkably attenuated with co-administration of various fractions and rutin thereby increased the level of CAT, POD, SOD, GSH, GSR, GST, GSH-Px, quinone reductase, while reduced the xanthine oxidase, gamma-GT, TBARS, H(2)O(2), nitrite, tissue proteins and DNA fragmentation%. Ameliorated effects of fractions and rutin were also recorded for the function of kidneys and the level of urobilinogen, urea, albumin, creatinine, RBC and WBC in urine were decreased. Serum level of creatinine, urobilinogen, BUN, direct bilirubin, total bilirubin and globulin were decreased while total proteins, albumin and creatinine clearance were increased with fractions and rutin. Protective effects of rutin and fractions were also evident on histopathology by reducing glomerular atrophy, tubular degeneration, congestion of blood capillaries, necrosis of epithelium and edema. Similarly body weight was increased while kidney and relative kidney weight was decreased with co-administration of fractions and rutin., Conclusion: It is suggested that Launaea procumbens effectively protect kidneys against the CCl(4)-induced oxidative damage in rats, through antioxidant and free radical scavenging effects of flavonoids and saponins present in the fractions., (Copyright (c) 2010 Elsevier Ireland Ltd. All rights reserved.)