Evaluation of Phytochemistry and Pharmacological Properties of Alnus nitida.

In the current study, the anti-inflammatory and analgesic potential of Alnus nitida (leaves and fruits) was evaluated in the Sprague-Dawley rat. Traditionally, A. nitida was used for the treatment of inflammatory ailments. However, A. nitida leaves and fruits have not been yet reported regarding any potential medicinal effects. Leaves/fruits of A. nitida were extracted with methanol and fractionated to attain n-hexane, chloroform, ethyl acetate and aqueous fractions. These extracts were then evaluated for in vivo analgesic and anti-inflammatory potential. For in vivo anti-inflammatory activity, carrageenan-induced paw edema assay, Freunds' complete adjuvant-induced edema, xylene-induced ear edema and histamine-induced paw edema models were used in rats, which showed significant (p < 0.01) reduction (70−80%) in edema in comparison of inflammatory controls. On other hand, for the analgesic assessment, hot plate assay and acetic acid-induced writhing tests were used, which showed a significant (p < 0.01) rise in latency time (40−60%) as compared with pain-induced controls. These results were comparable with standard drugs in a concentration-dependent manner and no mortality or toxicity was observed during all experiments. Then, for the identification of chemical constituents gas chromatography−mass spectrometry (GC-MS) analysis was performed, which indicated the presence of neophytadiene, 3,7,11,15-Tetramethyl-2-hexadecen-1-ol, phytol and vitamin E, justifying the use of A. nitida to treat inflammatory disorders.

Pharmacological evaluation of Acacia modesta bark for antipyretic, anti-inflammatory, analgesic, antidepressant and anticoagulant activities in Sprague Dawley rats.

In this study the bark of Acacia modesta was evaluated for anti-inflammatory, antipyretic, analgesic, antidepressant and anticoagulant activity by carrageenan, hot plat, forced swim and capillary tube method respectively in rats. Highest anti-inflammatory activity was exhibited by chloroform (AMC) extract (74.96% inhibition) while other two active fractions being n-hexane (AMH) and ethyl acetate (AME) exhibited 71.26% and 52.87% inhibition of edema respectively. On the other hand, the aqueous (AMA) fraction showed most effective response with 67.06% analgesic activity. Additionally, the significant (p<0.05) post-treatment antipyretic effect was found by all fractions in time dependent manner. The current findings showed that AMC, AME and AMA had significant reduction in immobility time in the antidepressant test, while AMH showed mild antidepressant activity. In anticoagulant assay, the coagulation time of crude extract A. modesta and its all fractions were comparable to that of positive control aspirin (208s). Moreover, neither mortality nor lethality was observed in the tested animals. Overall, the plant extracts showed potent anti-inflammatory, antipyretic, analgesic, antidepressant and anticoagulant activities which concludes that the bark of A. modesta have significant therapeutic potential.

Gonadoprotective ability of Vincetoxicum arnottianum extract against bisphenol A-induced testicular toxicity and hormonal imbalance in male Sprague Dawley rats.

Vincetoxicum arnottianum (Wight) of family Apocynaceae is a rich source of therapeutic alkaloids, phenolics and flavonoids. Study aims to evaluate the protective potential of methanol extract of Vincetoxicum arnottianum (VAM) on bisphenol A (BPA)-induced testicular toxicity in male Sprague Dawley rat. Quantitative analysis of VAM for total phenolic (TPC), total flavonoid (TFC) and total alkaloid content (TAC) along with HPLC analysis for polyphenolics was carried out. BPA-induced testicular toxicity was determined through analysis of antioxidant enzymes, DNA damages and testicular histopathology along with reproductive hormones in serum of rat. VAM was constituted of TFC (382.50 ± 1.67 µg GAE/mg), TPC (291.17 ± 0.82 µg RE/mg), TAC (16.5 ± 0.5%), ferulic acid (2.2433 µg/mg) and vanillic acid (2.1249 µg/mg). VAM co-administration to BPA-treated rats attenuated the toxic effects of BPA and restored the body and testis weights. Altered level of luteinizing hormone (LH), testosterone and follicle-stimulating hormone (FSH) in serum, and level of antioxidants (GSH, POD, CAT and SOD) and nitric oxide in testis tissues of BPA-induced toxicity were significantly restored by VAM. Histological and comet assay studies also sanctioned the protective potential of VAM in BPA-intoxicated rats. The presence of polyphenols and alkaloids might contribute towards the scavenging and ameliorative potential of VAM in testicular toxicity induced by BPA.

Vincetoxicum arnottianum ameliorate inflammation by suppressing oxidative stress and pro-inflammatory mediators in rat.

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-1ß, 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-1ß, 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.

Antidiabetic and antioxidant potential of Alnus nitida leaves in alloxan induced diabetic rats.

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 H2O2, 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.

Estimation of phytochemical constituents and in vitro antioxidant potencies of Brachychiton populneus (Schott & Endl.) R.Br.

BACKGROUND: Plants either in raw form or their isolated bioactive constituents are utilized as complementary and alternative medicine in various disorders. The present study was designed to evaluate chief phytochemical constituents of various fractions of Brachychiton populneus leaves and its antioxidative aptitude against free radicals. METHODS: Various fractions of B. populneus were prepared through solvent-solvent extraction technique based on their polarity and screened for phytochemical classes, total phenolic (TPC), flavonoid (TFC) and total tannin (TTC) content. Antioxidant effects of the extracts were manifested by in vitro multidimensional assays i.e. DPPH, hydroxyl radical scavenging, iron chelating, nitric oxide scavenging, ß-carotene bleaching, phosphomolybdenum and reducing power assay. RESULTS: Qualitative screening of various fractions of B. populneus ensured the presence of alkaloids, saponins, terpenoids, phenols, tannins, triterpenoids and flavonoids. Quantitative analysis revealed that aqueous fraction (BPA) showed maximum quantity of TPC and TFC followed by BPE and BPB. In terms of IC50 values BPA exhibited minimum values in all the in vitro antioxidant assays. However, the phytochemicals and yield did not accumulate in various fractions on polarity. CONCLUSION: Our results indicated the presence of various polyphenolics, flavonoids, alkaloids etc. The yield of various fractions and qualitative phytochemical analysis did not correlate with polarity of solvents. Various antioxidant assays exhibited significant (p < 0.05) correlation with TPC and TFC and renders B. populneus with therapeutic potential against free-radical-associated oxidative damages and this effect was significant with BPA.

Antioxidant, anti-inflammatory and hypoglycemic effects of Fagonia olivieri DC on STZ-nicotinamide induced diabetic rats - In vivo and in vitro study.

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-ß1, 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.

Potent anti-cancer activity of Alnus nitida against lung cancer cells; in vitro and in vivo studies.

Alnus nitida is used for multiple disorders in norther areas of Pakistan. In this study we have evaluated methanol extract of leaf (ANL) and stem bark (ANB) of A. nitida against two lung cancer cell lines; A-549 and H460 (Human non-small lung cancer cell lines) during in vitro assays for growth inhibition. Treatment with ANL and ANB markedly inhibited the growth of both cancer cell lines. Exposure of A-549 and H460 cancer cell lines to ANL and ANB inhibited cell survival, colony growth and migration of cells. Further, treatment of A-549 and H460 with ANL and ANB indicated alteration in actin fibers after staining with rhodamine-phalloidin. Both extracts cause shrinkage and cell cycle arrest during G1 phase. Treatment of A-549 and H460 cancer cells with ANL and ANB repressed the expression of anti-apoptotic proteins Bcl-2 and Bcl-xL along with downregulation of NFκB, cyclin D1 and PI3-K protein. In addition, intraperitoneal injection of ANL and ANB (10 mg/kg bw and 20 mg/kg bw) to C57BL/6 J mice implanted with B16F10 (Mouse melanoma cancer cell line) cells significantly (p < 0.01) decreased the number of nodules per lung and the level of various proteins reciprocating the in vitro studies. These results suggest that ANL and ANB be explored further for therapeutic use in lung cancer.

In vitro antioxidant efficacy and the therapeutic potential of Wendlandia heynei (Schult.) Santapau & Merchant against bisphenol A-induced hepatotoxicity in rats.

The aim of present study was to access the antioxidant and ameliorative efficacy of Wendlandia heynei stem bark's crude methanol extract (WHBM) against bisphenol A (BPA)-induced hepatotoxicity in the rat moel. WHBM and its derived fractions exhibited promising activity for the scavenging of DPPH, hydroxyl and nitrite radicals, iron chelation, and for the inhibition of ß-carotene oxidation. The administration of BPA to Sprague Dawley rats (25 mg kg-1) for 28 days resulted in an elevated (p < 0.01) level of aspartate transaminase, alanine transaminase, alkaline phosphatase, and globulin, and at the same time a decrease (p < 0.01) in the level of total protein and albumin in the serum of the rats. In hepatic samples, the levels of catalase, peroxidase, superoxide dismutase, glutathione-S-transferase, and reduced glutathione were decreased (p < 0.05), whereas thiobarbituric acid reactive substances, hydrogen peroxide, and the nitrite content were increased (p < 0.05) with BPA treatment to the rats. The administration of WHBM to BPA-intoxicated rats restored the altered levels of these parameters toward the control animals. Histopathological alterations of the hepatic tissues induced with BPA were restored with WHBM co-treatment to the rats. HPLC-DAD analysis ensured the occurrence of rutin, catechin, and caffeic acid in WHBM and WHBE. The results of this study suggested that the presence of phenolics and flavonoids in W. heynei bark might be responsible for it exhibiting antioxidant potential during the in vitro and in vivo studies and hence it has potential to be used as a therapeutic agent against oxidative stress associated diseases.

Phytochemical investigation and antimicrobial appraisal of Parrotiopsis jacquemontiana (Decne) Rehder.

BACKGROUND: Parrotiopsis jacquemontiana (Decne) Rehder. is locally used for skin infections and in wound healing. In this study we have evaluated methanol extract of its leaves and derived fractions against the clinical multi-drug resistant bacterial strains. METHODS: P. jacquemontiana leaves powder extracted with 95% methanol (PJM) and fractionated in escalating polarity of solvents; n-hexane (PJH), chloroform (PJC), ethyl acetate (PJE), n-butanol (PJB) and the remaining as aqueous fraction (PJA). Clinical as well as environmental 19 bacterial strains and 8 fungal strains were screened for minimum inhibitory concentration (MIC) and minimum bactericidal/fungicidal concentration (MBC/MFC). Preliminary phytochemical investigation for various phytochemical classes was also carried out. RESULTS: PJM contained the coumarins, phenols, flavonoids, tannins, alkaloids, glycosides, saponins, sterols, phlobatannins, steroids, phytosterols, triterpenoids, acids, quinones, proteins, vitamin C, betacyanins, oils and resins while anthraquinones, phytosteroids, carbohydrates and anthocyanins were not detected. Disc diffusion assay (1 mg/disc) indicated the sensitivity of all the MDR strains of bacteria with PJM, PJE and PJB, while no inhibition was recorded with PJA. PJH and PJC inhibited the growth of all the strains of Staphylococcus aureus, Pseudomonas aeruginosa and Coagulase negative staphylococci used in this study. Maximum zone of inhibition (35.5 ± 1.32 mm) was obtained with PJM against Staphylococcus lugdenesis MDR (6197). Comparatively lower MIC (8-64 µg/ml) and MBC (32-256 µg/ml) values were recorded for PJM and PJE. In case of fungal strains only PJM, PJE and PJB markedly inhibited the growth and lower MIC (8-128 µg/ml) and MFC (32-512 µg/ml) values were determined for PJM and PJE. CONCLUSION: The remarkable inhibition of various bacterial and fungal strains at low doses of the extract/fractions suggested the strong antibacterial, antifungal and anti-candidal potential of P. jacquemontiana leaves.

Evaluation of Artemisia scoparia for hemostasis promotion activity.

Excessive hemorrhage through any reasons is a life threatening process. Artemisia scoparia of family Asteraceae has been used in local system of medicine to stop bleeding from wounds and in injuries, antiseptic, in healing urticarial and for removal of worms from the body. Aerial parts of A. scoparia was extracted with 95% methanol (ASM) and fractionated through liquid-liquid partition in ascending order of n-hexane (ASH), chloroform (ASC), ethyl acetate (ASE), and the remaining as the aqueous fraction (ASA). Phytochemical classes of the extract/fractions were determined by qualitative assays. Prothrombin time (PT) was estimated on the plasma of human blood by Owren method. Capillary tube method was applied to determine the hemostasis activity in Sprague-Dawley rat. Tannins, saponins, terpenoids, quinones, betacyanins and flavonoids were present whereas phlobatannins, anthraquinones and alkaloids were established absent in ASM, ASC, ASE and ASA. Prothrombin time was significantly decreased by mixing (10 µg) of ASM (16.67±1.15 sec), ASH (12.33±0.57 sec), ASC (15.33±0.57 sec) and ASA (9.0±1.0 sec) to that of vehicle (20.0±1.0 sec). Administration (200 mg/kg) of all the extract/fractions showed significantly less (26.00±11.79 sec - 41.00±7.21 sec) hemostasis time as compared to the (242.67±39.67 sec) control rats. The results suggested the therapeutics importance of A. scoparia use in bleeding pathologies.

Protective potential of Parrotiopsis jacquemontiana (Decne) Rehder on carbon tetrachloride induced hepatotoxicity in experimental rats.

The current study aimed at exploring hepatoprotective potential of Parrotiopsis jacquemontiana leaves methanol extract (PJM) in rat. PJM constituted of significant amount of total phenolic and total flavonoid content. PJM and ethyl acetate fraction (PJE) exhibited strong antioxidant potential (DPPH, hydroxyl ion, nitric oxide, iron chelating, ß-carotene, reducing power and phosphomolybdate assay) in comparison to other extracts. CCl4 toxicity displayed significant (p<0.05) increase in level of TBARS, H2O2, nitrite while a decrease in SOD, CAT, POD and GSH in liver samples of CCl4 treated group. Serum analysis indicated significant increase in levels of ALT, AST and ALP and globulin whereas a decrease in albumin and total protein was also reported. In addition, levels of pro-inflammatory and fibrogenic cytokines (TNF-α, IL-6, TGF.ß1, and leptin) of liver serum were elevated whereas adiponectin (anti-inflammatory cytokine) level was decreased in treated rats. CCl4 induced DNA damage and histopathological architecture was also studied. However, co-administration with PJM in a dose-dependent manner reversed all the above pathological parameters caused by CCl4 toxicity. HPLC-DAD analysis confirmed the existence of rutin, gallic acid, myricetin and caffeic acid in PJM and PJE while catechin was additionally present in PJM. Based on rich-antioxidant profile and hepatoprotective outcome on various pathological parameters prompted by CCl4, it can be assumed to be a promising candidate shrub for phytomedicine development against various liver ailments and oxidative-stress related disorders.

Investigations on anti-inflammatory and analgesic activities of Alnus nitida Spach (Endl). stem bark in Sprague Dawley rats.

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.

Five Indigenous Plants of Pakistan with Antinociceptive, Anti-Inflammatory, Antidepressant, and Anticoagulant Properties in Sprague Dawley Rats.

Five medicinal plants of Pakistan were investigated for their antinociceptive, anti-inflammatory, antidepressant, and anticoagulant potential. Antinociceptive activity was estimated by hot plate and writhing assay. In hot plate assay, Quercus dilatata (52.2%) and Hedera nepalensis (59.1%) showed moderate while Withania coagulans (65.3%) displayed a significant reduction in pain. On the other hand, in writhing assay, Quercus dilatata (49.6%), Hedera nepalensis (52.7%), and Withania coagulans (62.0%) showed comparative less activity. In anti-inflammatory assays crude extracts showed significant edema inhibition in a dose dependent manner. In carrageenan assay, the highest activity was observed for Withania coagulans (70.0%) followed by Quercus dilatata (66.7%) and Hedera nepalensis (63.3%). Similar behavior was observed in histamine assay with percentage inhibitions of 74.3%, 60.4%, and 63.5%, respectively. Antidepressant activity was estimated by forced swim test and the most potent activity was revealed by Withania coagulans with immobility time 2.2s (95.9%) followed by Hedera nepalensis with immobility time 25.3s (53.4%). Moreover, the crude extracts of Fagonia cretica (74.6%), Hedera nepalensis (73.8%), and Phytolacca latbenia (67.3%) showed good anticoagulant activity with coagulation times 86.9s, 84.3s, and 67.5s, respectively. Collectively, the results demonstrate that these five plants have rich medicinal constituents which can be further explored.

Hepatoprotective potential of Fagonia olivieri DC. against acetaminophen induced toxicity in rat.

BACKGROUND: Fagonia olivieri (DC) being used for the treatment of diabetes, cancer, fever and claimed to be effective in many other stress related disorders. In this study we have evaluated the F. olivieri whole methanol extract and its derived fractions for various in vitro and in vivo antioxidant studies. METHODS: The crude methanol extract of the whole plant of F. olivieri (FOM) and its derived fractions; n-hexane (FOH), chloroform (FOC), ethyl acetate (FOE), n-butanol (FOB) and aqueous (FOA) were evaluated for the total phenolic and flavonoid content and in vitro antioxidant abilities. The antioxidant effect of FOM was determined by acetaminophen-induced hepatotoxicity in Sprague-Dawley (Rattus novergicus) male rats. The methanol/fractions were also analysed by HPLC analysis for the presence of polyphenolics. RESULTS: The total phenolic content of the samples ranged from 19.3 ± 0.529 to 106.2 ± 0.892 mg GAE/g extract while total flavonoid content 16.2 ± 0.881 to 50.1 ± 1.764 mg RTE/g extract, respectively. FOA showed highest radical scavenging activity for DPPH (IC50 = 55.2 ± 1.212 µg/ml), ABTS (IC50 = 90.2 ± 1.232 µg/ml) superoxide (IC50 = 37.1 ± 0.643 µg/ml) and for H2O2 (IC50 = 64 ± 1.463 µg/ml). FOE exhibited the highest antioxidant activities for phosphomolybdenum (IC50 = 78.2 ± 0.883 µg/ml) and for hydroxyl radical scavenging (IC50 = 82 ± 2.603 µg/ml). HPLC analysis of FOM and its derived fractions showed the presence of rutin, catechin and gallic acid. Elevated levels of AST, ALT, ALP, LDH and lipid profile in serum and lipid peroxidation and DNA damages in liver; while decreased activity level of CAT, SOD, GSH-Px, GR and reduced glutathione (GSH) concentration induced with acetaminophen in rat were reverted towards the control group with co-administration of FOM. CONCLUSION: Our results showed that F. olivieri is a potential source of natural antioxidants, which justifies its use in folklore medicine.

Protective effects of Fraxinus xanthoxyloides (Wall.) leaves against CCl4 induced hepatic toxicity in rat.

BACKGROUND: Leaves and root bark of Fraxinus xanthoxyloides Wall. (Oleaceae) are used locally for the treatment of jaundice, malaria and pneumonia. Decoction of stem, twigs and bark is used in pain, internal injuries, rheumatism and in bone fracture. In this investigation we have evaluated the methanol extract of leaves for its hepatoprotective potential against CCl4 induced hepatic injuries in rat. METHODS: Powder of F. xanthoxyloides leaves was extracted with methanol (FXM) and subjected for the determination of polyphenolics through HPLC-DAD analysis. Sprague-Dawley (Rattus novergicus) male rats were divided into eight groups (six rats in each). Group I: non-treated control; Group II: vehicle treated (DMSO plus olive oil) while Group III- VI treated with 1 ml/kg body weight (bw) of CCl4 (30 % in olive oil) for 30 days (15 dosages) to induce the hepatic injuries. Group IV: treated with silymarin (100 mg/kg bw); Group V and VI with FXM (200, 400 mg/kg bw) on alternate days with CCl4 treatment. Group VII and VIII was administered with FXM (200, 400 mg/kg bw) alone (15 dosages). After 30 days the serum was evaluated for liver function enzymes and biochemical markers, liver samples for antioxidant enzymes, biochemical markers, comet assay and for histopathology. RESULTS: HPLC-DAD analysis of FXM revealed the existence of rutin and caffeic acid. In CCl4 treated rats the level of alanine transaminase (ALT), aspartate transaminase (AST), total bilirubin was significantly increased while the albumin concentration in serum was decreased as compared to control group. The level of hepatic antioxidant enzymes, catalase (CAT), peroxidase (POD), superoxide dismutase (SOD), glutathione-S-transferase (GST) and glutathione reductase (GSR) was significantly decreased against the control group. Further, significant decrease in GSH while increase in lipid peroxides (TBARS), H2O2, DNA damages and comet length was induced with CCl4 in hepatic tissues of rat. In contrast, co-administration of FXM and silymarin restored the biochemical and histopathological status of the liver. CONCLUSION: Results of present investigation revealed that F. xanthoxyloides leaves possibly protect the liver against CCl4 induced injuries like silymarin by its antioxidant constituents.

Phytochemical, antioxidant and hepatoprotective effects of Alnus nitida bark in carbon tetrachloride challenged Sprague Dawley rats.

BACKGROUND: Alnus nitida (Spach) Endl. is traditionally used for inflammatory disorders. Diarylheptanoids constituents having diverse therapeutically importance including hepato-protective was reported in A. nitida. The aim of this study was to explore the antioxidant and hepato-protective profile of A. nitida stem bark's crude methanol extract (ANM). METHODS: Crude methanol extract of A. nitida stem bark and its derived fractions were assessed for phytochemical classes and in vitro antioxidant profiling by multidimensional assays. Hepato-protective assessment of ANM was investigated on rats, which were made hepatotoxic using carbon tetrachloride (CCl4). Additionally HPLC-DAD analysis of ANM, and its derived ethyl acetate and aqueous fraction was carried out to determine the presence of active constituents. RESULTS: Qualitative analysis of crude extract-and its fractions depicted the presence of terpenoids, saponins, coumarins, phenols and flavonoids. Maximum quantity of total phenolic content (TPC) and total flavonoid content (TFC) was recorded in ANM and its derived fractions; n-hexane (ANH), chloroform (ANC), ethyl acetate (ANE) and the residual aqueous (ANA). ANM exhibited the best total antioxidant capacity, total reducing power, and scavenging of DPPH and OH radicals. ANE and ANA exhibited strong scavenging potential for iron chelation, nitric oxide and ß-carotene bleaching assay. ANM treatment converse the activities of serum-marker enzymes and lipid profile, altered by CCl4 treatment in rat. CCl4 induced hepatic-cirrhosis in rat resulted in decrease of antioxidant enzyme activities such as catalase, peroxidase, superoxide dismutase, glutathione peroxidase, glutathione-S-transferase and glutathione reductase-which were restored towards the normal level with ANM. Similarly diminished level of reduced glutathione while enhanced level of lipid peroxides, hydrogen peroxide and nitrite in liver of cirrhotic rats was normalized by treatment of ANM. The histopathological studies of liver tissues also represented that ANM possessed the hepato-protective activity. HPLC-DAD analysis against eight known standards confirmed the presence of gallic acid, catechin and rutin in ANM and in ANA while in ANE gallic acid was only detected. CONCLUSION: Based on the results of antioxidants, restoration of various antioxidant enzymes and histopathological studies, the recent study concludes that antioxidant potential of A. nitida bark might protect the liver damages.

Fraxinus xanthoxyloides leaves reduced the level of inflammatory mediators during in vitro and in vivo studies.

BACKGROUND: Different parts of Fraxinus xanthoxyloides Wall. (Oleaceae) are used traditionally in the treatment of internal wounds, bone fracture, pain, jaundice, malaria and in pneumonia. These ailments involve protective and essential mechanisms of the organism in response to infection, injury and trauma. However, prolonged inflammation may lead to inflammatory disorders. The present investigation was carried to evaluate the crude methanol extract of F. xanthoxyloides leaves and its fractions for their anti-inflammatory and analgesic effects. METHODS: Methanol extract of F. xanthoxyloides leaves was fractionated through liquid-liquid partition on escalating polarity of solvents. Acetic acid and thermal responses were used to evaluate the analgesic effects of extract/fractions in rat. Anti-inflammatory effects were monitored through in vitro; TNF-α activated NFkB in 293/NFkB-Luc HEK cells and LPS-activated nitric oxide (NO) assay in RAW 264.7 cells. For in vivo studies carrageenan induced paw edema model was used in rat. Both in vitro and in vivo studies have indicated that chloroform fraction exhibited superior anti-inflammatory effects to other extract/fractions and therefore, was used in air pouch model in rat to estimate the inhibition in leukocyte migration and synthesis of inflammatory mediators. In addition, phytochemical investigation of crude extract was carried out by GC-MS analysis. RESULTS: GC-MS studies of crude extract revealed the presence of various classes of which terpenoids (26.61 %), lactam (16.47 %), esters (15.81 %), phenols (8.37 %), and steroid (6.91 %) constituted the major categories. Among the extracts chloroform fraction (200 mg/kg bw) significantly (P <0.001) increased the percent latency time (76.13 ± 4.49 %) in hot plate test after 120 min and decreased (P <0.001) the count of writhes (77.23 ± 5.64 %) as compared to other extracts. The in vitro studies indicated that chloroform fraction at 15 µg/ml more effectively inhibited the TNF-α induced synthesis of NFkB (85.0 ± 8.12 %, IC50 = 5.98 µg/ml) and LPS-instigated nitric oxide (78.23 ± 2.39 %, IC50 = 6.59 µg/ml) synthesis. Although all the extract/fractions showed a dose dependent increase in inhibition of edema formation however, chloroform fraction (4(th) h = 77.64 ± 3.04 %) at 200 mg/kg bw exhibited relatively higher (P <0.001) anti-inflammatory activity in carrageenan-induced paw edema in rat. Moreover, chloroform fraction had the ability to decrease (P <0.001) the influx of leukocytes and the concentration of inflammatory mediators; TNF-α, NO, IL-6 and PGE2 in air pouch exudate. CONCLUSION: The study demonstrates the therapeutic potential of F. xanthoxyloides leaves against the inflammatory disorders suggesting the presence of active constituents in chloroform fraction.

Proficiencies of Artemisia scoparia against CCl4 induced DNA damages and renal toxicity in rat.

BACKGROUND: Artemisia scoparia is traditionally used in the local system of medicine in kidney disorders. This study aimed at scrutinizing the nephroprotective prospective of A. scoparia methanol extract against carbon tetrachloride (CCl4) provoked DNA damages and oxidative stress in kidneys of rat. METHODS: Dried aerial parts of A. scoparia were powdered and extracted with methanol to obtain the viscous material (ASM). Sprague Dawley male rats (42) were grouped (7) having 6 rats in each. Group I remained untreated and Group II treated intraperitoneally (i.p) with DMSO + olive oil (1 ml/kg body weight (bw). Rats of Group III - VI were treated with CCl4 (1 ml/kg bw; i.p 30 % v/v in olive oil). Animals of Group IV were co-administered with 100 mg/kg bw of silymarin whereas rats of Group V and VI with 150 mg/kg bw and 300 mg/kg bw of ASM at an interval of 48 h for four weeks. Animals of Group VII were administered with ASM (300 mg/kg bw) alone. DNA damages were investigated with comet assay in renal tissues while the oxidative injuries were estimated in serum and renal tissues. RESULTS: Co-administration of ASM to rats significantly reduced the DNA damages at 300 mg/kg dose as indicated in comet length (40.80 ± 2.60 µm), head length (34.70 ± 2.21 µm), tail length (7.43 ± 1.24 µm) and DNA content in head (88.03 ± 2.27 %) to that of CCl4 for comet length (63.16 ± 2.11 µm), head length (23.29 ± 1.50 µm), tail length (39.21 ± 2.81 µm) and DNA content of head (74.81 ± 2.18 %) in renal cell's nuclei. Increased level of urea, creatinine, bilirubin, blood urea nitrogen whereas decreased concentration of proteins in serum of CCl4 treated animals were restored towards the normal level with co-administration of ASM. CCl4 injection in rats decreased the activity level of CAT, POD, SOD, GST and γ-GT and GSH contents while elevated levels of TBARS, H2O2 and nitrite contents were observed in renal tissues. A noteworthy retrieval of all these parameters and the altered histopathological observations was notified near to the normal values after treatment with both the doses of ASM. CONCLUSION: Results obtained suggested the therapeutic role of ASM in oxidative stress related disorder of kidneys.

Transformation of Lactuca sativa L. with rol C gene results in increased antioxidant potential and enhanced analgesic, anti-inflammatory and antidepressant activities in vivo.

Lettuce is an important edible crop which possesses various medicinal properties. In this study Lactuca sativa L. (cv Grand Rapids) was transformed by Agrobacterium-mediated transformation with rol C gene. Transgene integration and expression was confirmed through PCR and semiquantitative RT-PCR. The transformed extracts were evaluated for their in vitro antioxidant and in vivo analgesic, anti-inflammatory and antidepressant activities in rats. The transformed plants showed 53-98 % increase in total phenolic and 45-58 % increase in total flavonoid contents compared with untransformed plants. Results of total reducing power and total antioxidant capacity exhibited 90-118 and 61-75 % increase in transformed plants, respectively. In contrast to control, DPPH, lipid peroxidation and DNA protection assay showed up to 37, 20 and 50 % enhancement in transformed plants, respectively. The extracts showed similar but significant enhancement behavior in hot plate analgesic and carrageenan-induced hind paw edema test. The transformed extracts showed 72.1 and 78.5 % increase for analgesic and anti-inflammatory activities, respectively. The transformants of rol C gene exhibited prominent antidepressant activity with 64-73 % increase compared with untransformed plants. In conclusion, the present work suggests that transformation with rol C gene can be used to generate lettuce with enhanced medicinally important properties, such as antioxidant, analgesic, anti-inflammatory and antidepressant potential.