Ameliorative effect of methanol extract of Rumex vesicarius on CCl4-induced liver damage in Wistar albino rats

Abstract

Context: Rumex vesicarius L. (Polygonaceae), an edible plant, is reported to have many bioactive phytochemicals, especially flavonoids and anthraquinones with antioxidant and detoxifying properties.

Objective: This study evaluated the methanolic extract of R. vasicarius (MERV) for hepatoprotective activity in rats against CCl₄-induced liver damage.

Materials and methods: The whole plant extract was prepared and investigated for its hepatoprotective activity. Rats were pretreated with MERV (100 and 200?mg/kg, p.o.) for 7?d prior to the induction of liver damage by CCl₄. Animals were then sacrificed 24?h after CCl₄ administration for the biochemical (AST, ALT, and ALP activity in serum; lipid peroxidation (LPO) and glutathione (GSH) levels in liver tissue) and histological analyses.

Results: CCl₄-induced hepatotoxicity was confirmed by an increase (p?<?0.05) in serum AST (4.55-fold), ALT (3.51-fold), and ALP (1.82-fold) activities. CCl₄-induced hepatotoxicity was also manifested by an increase (p?<?0.05) in LPO (3.88-fold) and depletion of reduced glutathione (3.14-fold) activity in liver tissue. The multiple dose MERV administration at 200?mg/kg showed promising hepatoprotective activity as evident from significant decrease levels of serum AST (230.01?±?13.21), serum ALT (82.15?±?5.01), serum ALP (504.75?±?19.72), hepatic LPO (3.38?±?0.33), and increased levels of hepatic glutathione (0.34?±?0.04) towards near normal. Further, biochemical results were confirmed by histopathological changes as compared with CCl₄-intoxicated rats.

Discussion and conclusion: The results obtained from this study indicate hepatoprotective activity of Rumex plant against CCl₄-induced liver toxicity; hence, it can be used as a hepatoprotective agent.     

Effect of hydroalcoholic extract of Aegle marmelos fruit on radical scavenging activity and exercise-endurance capacity in mice

Context: Aegle marmelos L. Corr (Rutaceae) is an important Indian Ayurvedic medicinal plant used for the treatment of various ailments. However, little information is available on the anti-fatigue properties of its fruit.

Objective: Evaluation of the physical endurance and exercise-induced oxidative stress modulating properties of A. marmelos fruit in mice.

Material and methods: Radical scavenging activity of the fruit hydroalcoholic extract was evaluated using in vitro systems. The extract was further evaluated for its endurance-enhancing properties at three oral doses (100, 200 and 400?mg/kg?b.wt) in BALB/c mice for 21?d using a swimming test.

Results and discussion: The extract exhibited significant scavenging activity against DPPH (IC₅₀, 351?±?37?µg/ml) and ABTS radicals (IC₅₀, 228?±?25?µg/ml), respectively, with the polyphenol content of 95?µg/mg extract. It also inhibited AAPH radical-induced oxidation of biomolecules such as BSA protein (63%), plasmid DNA (81%) and lipids (80.5%). Administration of extract resulted in an increase in the duration of swimming time to exhaustion by 23.4 and 47.5% for medium and higher doses, respectively. The extract significantly normalized the fatigue-related biochemical parameters and also down-regulated the swim stress-induced over-expression of heat shock protein-70 and up-regulated the skeletal muscle metabolic regulators (GLUT-4 and AMPK1-α) by 2- and 3-fold, respectively, at the higher dose in muscle tissues.

Conclusion: Our study demonstrates the anti-fatigue properties of A. marmelos fruit, most probably manifested by delaying the accumulation of serum lactic acid, increasing the fat utilization and up-regulating the skeletal muscle metabolic regulators.     

Ameliorative effect of Aegle marmelos leaf extract on early stage alloxan-induced diabetic cardiomyopathy in rats

Objective: The pathogenesis of diabetic cardiomyopathy (DCM) is complex, and the therapeutic options available to treat DCM are limited. The present study was designed to investigate the effect of Aegle marmelos (L.) Correa (Rutaceae) leaf extract on early stage DCM in alloxan-induced diabetic rats.

Methods: Diabetes was induced in Wistar rats (150–200?g) by injecting alloxan (150?mg kg^?1; i.p.). Ethanol extract of A. marmelos leaves was administered at varying doses (100, 200, and 400?mg kg^?1) and tolbutamide (100?mg kg^?1) as standard. Fasting blood glucose (FBG), total cholesterol, thiobarbituric acid reactive substances (TBARS), reduced glutathione (GSH), catalase (CAT), superoxide dismutase (SOD), lactate dehydrogenase (LDH) and creatine kinase (CK) were determined by standard methods.

Results: A. marmelos extract (AME) was found to decrease the levels of FBG, total cholesterol, TBARS, LDH and CK, and increase the levels of GSH, CAT and SOD dose dependently as compared to diabetic control groups. The maximum dose-dependent decrease in TBARS (63.46%), LDH (34.04%), CK (53.14%), and increase in GSH (64.91%), CAT (59.34%), SOD (69.65%) was evident at an optimum dose of 200?mg kg^?1. Histopathological studies revealed salvage in the morphological derangements as indicated by absence of necrosis and marked decrease in inflammatory cells in AME-treated groups as compared to diabetic control.

Conclusions: The present investigations conclude that treatment with AME attenuates the severity and improves the myocardium in the early stages of alloxan-induced DCM at a dose of 200?mg kg^?1.     

Protective Effect of Aegle marmelos Fruit in Gastrointestinal Dysfunction in rats

The effect of the ethanol extract of the unriped fruits of Aegle marmelos Correa was assessed on experimentally induced diarrhoea and gastric ulceration in rats. The extract (50, 100 and 200?mg/kg, p.o.) exhibited a dose-dependent decrease in the intestinal propulsion from 61.79–39.32% which is equivalent to 38.21–60.68% intestinal propulsion inhibition (control 58.3 ± 3.4 inhibition, P < 0.5 to P < 0.001) and caused a dose-dependent decrease in the total number of faecal matter in castor oil-induced diarrhoea (control 70, reduced to 51 and 42 at 100 and 200?mg/kg extract, p.o.). Further, yohimbine, a α₂ adrenoreceptor blocker, attenuated the antidiarrhoeal effect of the extract in a dose of 200?mg/kg to 17.14%, and diphenoxylate by 74.28%. The extract also reduced the ulcer index induced by ethanol (control 18.7 ± 4.4, 34.22–72.73% protection), aspirin (control 22.6 ± 3.4, 36.73–81.42% protection) and cold restraint stress (control 23.8 ± 3.2, 56.72% and 81.51% protection). Further study on tissue lipid peroxidation was significantly increased (P < 0.001) as evidenced by accumulation of malondialdehyde in cold restraint stress ulcers. Administration of A. marmelos (100 and 200?mg/kg), cimetidine 50?mg/kg and reduced glutathione (150?mg/kg) prior to cold restraint stress causes significant decrease in ulcer index and lipid peroxidation (P < 0.01 to P < 0.001). The result showed that A. marmelos had significant antidiarrhoeal and ulcer protective activity by scavenging the reactive oxygen species on the cold restraint stress-induced gastric damage.     

Antioxidant and hepatoprotective effects of Solanum xanthocarpum leaf extracts against CCl4-induced liver injury in rats

Context: Solanum xanthocarpum Schard. and Wendl. (Solanaceae) has been used in traditional Indian medicines for its antioxidant, anti-inflammatory, and antiasthmatic properties.

Objective: The present study demonstrates the antioxidant and hepatoprotective effects of S. xanthocarpum. On the basis of in vitro antioxidant properties, the active fraction from column chromatography of the methanol extract of S. xanthocarpum leaves (SXAF) was chosen as the potent fraction and used for hepatoprotective studies in rats.

Materials and methods: The antioxidant activity was evaluated by 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and reducing power assays. Rats were pre-treated with 100 and 200?mg/kg b.w. of SXAF for 14?d with a single dose of CCl₄ in the last day. Hepatoprotective properties were determined by serum biochemical enzymes, aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), antioxidant enzymes (SOD, CAT, GSH, and GST), and histopathology studies.

Results: SXAF exhibited significant antioxidant activity in scavenging free radicals with IC₅₀ values of 11.72?µg (DPPH) and 17.99?µg (ABTS). Rats pre-treated with SXAF demonstrated significantly reduced levels of serum LDH (1.7-fold), ALP (1.6-fold), and AST (1.8-fold). Similarly, multiple dose SXAF administration at 200?mg/kg b.w. demonstrated significantly enhanced levels of SOD (1.78?±?0.13), CAT (34.63?±?1.98), GST (231.64?±?14.28), and GSH (8.23?±?0.48) in liver homogenates. Histopathological examination showed lowered liver damage in SXAF-treated groups.

Discussion and conclusion: These results demonstrate that SXAF possesses potent antioxidant properties as well as hepatoprotective effects against CCl₄-induced hepatotoxicity.     

Phytoextract of Indian mustard seeds acts by suppressing the generation of ROS against acetaminophen-induced hepatotoxicity in HepG2 cells

Abstract

Context: Indian mustard [Brassica juncea (L.) Czern. & Coss. (Brassicaceae)] is reported to possess diverse pharmacological properties. However, limited information is available concerning its hepatoprotective activity and mechanism of action.

Objective: To study the protective mechanism of mustard seed extract against acetaminophen (APAP) toxicity in a hepatocellular carcinoma (HepG2) cell line.

Materials and methods: Hepatotoxicity models were established using APAP (2.5–22.5?mM) based on the cytotoxicity profile. An antioxidant-rich fraction from mustard seeds was extracted and evaluated for its hepatoprotective potential. The mechanism of action was elucidated using various in vitro antioxidant assays, the detection of intracellular generation of reactive oxygen species (ROS), and cell cycle analysis. The phytoconstituents isolated via HPLC-DAD were also evaluated for hepatoprotective activity.

Results: Hydromethanolic seed extract exhibited hepatoprotective activity in post- and pre-treatment models of 20?mM APAP toxicity and restored the elevated levels of liver indices to normal values (p?<?0.05). Post-treatment suppressed the generation of ROS by 58.37% and pre-treatment effectively prevented the generation of ROS by 90.5%. The mechanism of ROS suppression was further supported by antioxidant activity (IC₅₀) data from DPPH (103.37?±?4.2?µg AAE/mg), FRAP (83.26?±?1.1?µg AAE/mg), ORAC (1115?µM GAE/ml), ABTS (83.05?µg GAE/ml), and superoxide (345.22?±?5.15?µg AAE/mg) scavenging assays and by the restoration of cell cycle alterations. HPLC-DAD analysis revealed the presence quercetin, vitamin E, and catechin, which exhibited hepatoprotective activity.

Discussion and conclusions: A phytoextract of mustard seeds acts by suppressing the generation of ROS in response to APAP toxicity.     

Hepatoprotective and antioxidant effect of ellagitannins and galloyl esters isolated from Melaleuca styphelioides on carbon tetrachloride-induced hepatotoxicity in HepG2 cells

Context In a previous study, the total extract of Melaleuca styphelioides Sm. (Myrtaceae) showed a significant hepatoprotective effect in a CCl₄-induced toxicity model in mice. However, the active components responsible for the activity of the extract were not identified.

Objective To determine the in vitro hepatoprotective activity of the isolated pure compounds from M. styphelioides leaves using the CCl₄-challenged HepG2 cell model.

Materials and methods The hepatoprotective activity of the compounds (at concentrations of 100, 50 and 25?μm), the total extract and silymarin (Sil) (100, 50 and 25?μg/ml) was determined by measuring the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) after pretreatment with the tested samples for one hour. Glutathione (GSH) and superoxide dismutase activity (SOD) were estimated to determine the mechanisms of the hepatoprotective activity.

Results Some compounds showed marked hepatoprotection, including tellimagrandin I, which produced 42, 36 and 31% decrease in ALT and 47, 43 and 37% decrease in AST, at the tested concentrations, respectively, pedunculagin (32, 32 and 30% decrease for ALT and 48, 48 and 45% for AST), tellimagrandin II (38, 32 and 26% decrease for ALT and 45, 40 and 34% for AST) and pentagalloyl glucose (30, 28 and 26% decrease for ALT and 45, 38 and 36% for AST). Tellimagrandin I and II showed the highest increase in GSH (113, 105 and 81% and 110, 103 and 79%, respectively), which was comparable to Sil. Pedunculagin produced the highest increase in SOD (497, 350 and 258%).

Conclusion This study highlights promising natural hepatoprotective candidates derived from M. styphelioides.     

Protection of SAL B with H9C2 cells

Context: Salvianolic acid B (Sal B) is regarded as a potent antidiabetic agent and has been reported to possess cardioprotective effect in vivo.

Objective: This study investigated the cardioprotective effects of Sal B on H9c2 cells injury caused by high glucose in vitro, and clarified the possible mechanisms.

Materials and methods: Di ferent concentrations of Sal B were incubated with cells for 12?h prior being exposed to high glucose for 24?h. Cardioprotective effects of Sal B were evaluated using CCK-8 assay, ELISA, Hoechst 33258 nucleus staining, and western blot.

Results: Following a 24?h exposure of H9c2 to high glucose, obvious reduction was found in cell viability (45%), GSH (54.8?±?9.4?ng/mg protein), catalase (1.22?±?0.12 U/mg protein), and GPX level (67.9?±?9.4 U/mg protein), which were associated with the increases of GSSG (1.99?±?0.28?ng/mg protein) and ROS (2.00?±?0.19 RFU/mg protein) production. High glucose also elevated IL-6 (1.8-fold), IL-1β (1.9-fold), and TNF-α (1.6-fold) level, as well as induced cell apoptosis and NF-κB (6.1-fold) activation. However, Sal B (25 and 50?μM) elevated cell viability (28% and 44%), ameliorated oxidative stress (GSH, 1.3- and 1.6-fold; catalase, 1.9- and 2.0-fold; GPX, 1.1- and 1.4-fold; GSSG, 0.9- and 0.8-fold; ROS, 0.6- and 0.5-fold), and inflammatory response (IL-6, 0.9- and 0.7-fold; IL-1β, 0.8- and 0.6-fold; TNF-α, 0.9- and 0.8-fold), and inhibited cell apoptosis and NF-κB (0.5- and 0.2-fold) expression.

Conclusion: Sal B attenuated high glucose-induced injury and cytotoxicity through inhibiting inflammatory cytokine production in H9c2 cardiac cells.     

Anti-inflammatory activity of standardized dichloromethane extract of Salvia connivens on macrophages stimulated by LPS

Context: A previous study demonstrated that the chloroform extract of Salvia connivens Epling (Lamiaceae) has anti-inflammatory activity.

Objective: Identification of the active components in the dicholorometane extract (DESC), and, standardization of the extract based in ursolic acid.

Material and methods: DESC was prepared by percolation with dichlromethane and after washed with hot hexane, its composition was determined by CG-MS and NMR, and standardized by HPLC. The anti-inflammatory activity was tested on acute TPA-induced mouse ear oedema at doses of 2.0?mg/ear. The cell viability of macrophages was evaluated by MTT method, and pro- and anti-inflammatory interleukin levels were measured using an ELISA kit.

Results: Ursolic acid, oleanolic acid, dihydroursolic acid and eupatorin were identified in DESC, which was standardized based on the ursolic acid concentration (126?mg/g). The anti-inflammatory activities of DESC, the acid mixture, and eupatorin (2?mg/ear) were 60.55, 57.20 and 56.40% inhibition, respectively, on TPA-induced ear oedema. The IC₅₀ of DESC on macrophages was 149.4?μg/mL. DESC (25?μg/mL) significantly reduced TNF-α (2.0-fold), IL-1β (2.2-fold) and IL-6 (2.0-fold) in macrophages stimulated with LPS and increased the production of IL-10 (1.9-fold).

Discussion: Inflammation is a basic response to injuries, and macrophages are involved in triggering inflammation. Macrophage cells exhibit a response to LPS, inducing inflammatory mediators, and DESC inhibits the biosynthesis of the pro-inflammatory and promote anti-inflammatory cytokines.

Conclusion: DESC has an anti-inflammatory effect; reduced the levels of IL-1β, Il-6 and TNF-α; and increases IL-10 in macrophages stimulated with LPS. Ursolic acid is a good phytochemical marker.     

Antioxidative burst and hepatoprotective effects of ethanol root extract of Hippocratea africana against paracetamol-induced liver injury

Abstract

Context: Hippocratea africana (Willd.) Loes. ex Engl. (Celastraceae) root is used traditionally as an antipoison or antidote to treat liver diseases.

Objective: To evaluate antioxidative burst and hepatoprotective potentials of H. africana against paracetamol-induced liver injury in rats.

Materials and method: Antioxidative burst activity of the extract (1–100?µg/ml) in whole blood, neutrophils and macrophages was investigated using a luminol/lucigenin-based chemiluminescence assay. The hepatoprotective effect of the extract (200–600?mg/kg) was evaluated by the assay of liver function parameters, antioxidant enzymes and histopathological studies of the liver. GC-MS analyses of hexane and dichloromethane fractions were also carried out.

Results and discussion: The root extract/fractions exerted pronounced inhibition of oxidative burst activity in whole blood, neutrophils (intracellular and extracellular) and macrophages (3.04–99.70%). The administration of the root extract caused significant (p?<?0.05–0.001) reduction of high levels of liver enzymes (AST, ALT and ALP), total cholesterol, direct and total bilirubin as well as elevation of serum levels of total protein, albumin and antioxidant enzymes (SOD, CAT, GPx and GSH). Histology of the liver sections of extract and silymarin-treated animals showed reductions in the pathological features compared to the paracetamol-treated animals. The chemical pathological changes were consistent with histopathological observations suggesting a marked hepatoprotective effect of the root extract of H. africana. The GC-MS analysis revealed some pharmacologically active compounds.

Conclusion: The results show that the root extract of H. africana has hepatoprotective potential probably due to its antioxidative burst activity.     

Hepatoprotective effect of Commiphora myrrha against d-GalN/LPS-induced hepatic injury in a rat model through attenuation of pro inflammatory cytokines and related genes

Abstract

Context: Commiphora myrrha (Burseraceae), a shrub resembling a small tree, has been used for several centuries for the treatment of various diseases.

Objective: This study investigates the hepatoprotective activity of C. myrrha ethanol extract against d-galactosamine/lipopolysaccharide (d-GalN/LPS)-induced acute hepatic injury in an animal model.

Materials and methods: Rats were pretreated with ethanolic extract C. myrrha (250 and 500?mg/kg; p.o.) for 7 d prior to the induction of an acute phase response by d-GalN/LPS. Animals were sacrificed 24?h after d-GalN/LPS (800?mg/kg and 50?µg/kg i.p.) administration for the biochemical and histological analyses.

Results: The administration of d-GalN/LPS increased plasma aminotransferases (174.47?±?4.5761 and 260.96?±?1.9839?µkat/l) and total bilirubin levels (1.012?±?0.0288?mg/dl), which were attenuated by C. myrrha treatment. Hepatic lipid peroxidation activity and nitric oxide content also increased, while the antioxidant activity measured by GSH (0.76 nmol/g protein), SOD (81.91?U/mg protein), and CAT (15.78?U/mg protein) was reduced. Commiphora myrrha provided significant restoration of GSH (0.815 nmol/gm protein), SOD (140.57?U/mg protein), and CAT (27.02?U/mg protein) levels. Furthermore, the acute phase response elicited by d-GalN/LPS administration enhanced mRNA expressions of TNF-α, IL-6, IL-10, iNOS-2, and HO-1, which were ameliorated by C. myrrha treatment.

Discussion and conclusion: These findings indicate that C. myrrha considerably reduces the oxidative stress of d-GalN/LPS-induced hepatic injury via multiple pathways including adown regulation of inflammatory mediators and cytokines. Such a property might be sufficient to combat cellular damage caused by various conditions that resemble fulminant hepatitis and could be of a potential clinical application.     

Evaluation of antioxidant and hepatoprotective effects of white cabbage essential oil

Context: There have been no reports of the extraction of essential oil (EO) from white cabbage [Brassica oleracea L. var. capitata (L.) Alef. f. alba DC. (Brassicaceae)] (Bocfal) or its chemical composition, antioxidant activity, or hepatoprotective effects.

Objective: To extract Bocfal EO, to identify and quantify its chemical components, to assess their antioxidant capacity, and to evaluate the hepatoprotective properties of Bocfal EO.

Materials and methods: Bocfal EO was obtained using hydrodistillation (200?mm Hg/58?°C). The chemical composition was analyzed using GC-MS and was quantified using GC-FID. The antioxidant activity of Bocfal EO and its main constituents was evaluated using TBARS in rat brain homogenates. A Bocfal EO hepatoprotective effect (192?mg/kg) on acute carbon tetrachloride (CT)-induced liver damage was determined in rats using biochemical markers and histological analysis. Diallyl disulphide (DADS) (1?mmol/kg) was used as a control for comparison.

Results: Bocfal EO contained organic polysulphides (OPSs), such as dimethyl trisulphide (DMTS) 65.43?±?4.92% and dimethyl disulphide (DMDS) 19.29?±?2.16% as major constituents. Bocfal EO and DMTS were found to be potent TBARS inhibitors with IC₅₀ values of 0.51 and 3?mg/L, respectively. Bocfal EO demonstrated better hepatoprotective properties than did DADS (p?per se, as observed using histopathology.

Discussion and conclusion: The antioxidant properties of Bocfal EO and DMTS may be the mechanism of hepatoprotective action; the parenchymal disturbances by Bocfal EO or DADS alone may be related to the high doses used.     

Effect of Aegle marmelos on DEN initiated and 2-AAF promoted hepatocarcinogenesis: a chemopreventive study

In this study, we examined the inhibitory effects of Aegle marmelos methanolic extract on diethylnitrosamine (DEN) initiated and 2-acetyl aminofluorene (2-AAF) promoted liver carcinogenesis in male Wistar rats. Interestingly, it was found that A. marmelos (25 and 50?mg/kg body weight) resulted in a marked reduction of the incidence of liver tumors, which was further confirmed with histopathology. Furthermore to understand the underlying mechanisms of chemoprevention potential of A. marmelos, we evaluated the levels of hepatic antioxidant defence enzymes, ornithine decarboxylase (ODC) activity and hepatic DNA synthesis as a marker for tumor promotion since a direct correlation between these marker parameters and carcinogenicity have been well documented. Treatment of male Wistar rats for five consecutive days with 2-AAF induced significant hepatic toxicity, oxidative stress and hyper-proliferation. Pretreatment of A. marmelos extract (25 and 50?mg/kg body weight) prevented oxidative stress and toxicity by restoring the levels of antioxidant enzymes at both the doses. The promotion parameters (ODC activity and DNA synthesis) induced by 2-AAF administration in diet with partial hepatectomy (PH) were also significantly suppressed dose-dependently by A. marmelos. Therefore, we can conclude that ultimately the protection against liver carcinogenesis by A. marmelos methanolic extract might be mediated by multiple actions, which include restoration of cellular antioxidant enzymes, detoxifying enzymes, ODC activity and DNA synthesis.     

Boehmeria nivea attenuates LPS-induced inflammatory markers by inhibiting p38 and JNK phosphorylations in RAW264.7 macrophages

Abstract

Context: Boehmeria nivea (Linn.) Gaudich (Urticaceae), a natural herb, has a long history of treating several diseases including wound healing. However, the anti-inflammatory effect of B. nivea has not been investigated.

Objective: We investigated whether the 70% ethanol extract of B. nivea (Ebn) can exert anti-inflammatory activity. Several phenolic compounds of extracts were determined to provide further information on the correlation between anti-inflammatory effects and phenolic compounds.

Materials and methods: We prepared a 70% ethanol extract of B. nivea leaves and evaluated its anti-inflammatory activity (200, 400, 800, 1200?µg/mL) by measuring the secretions of nitric oxide (NO), tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6), which were stimulated by lipopolysaccharide (LPS) in RAW264.7 macrophages. The total phenolic compounds were determined by the Folin–Ciocalteu method and major compounds were determined by HPLC.

Results: Ebn was able to abolish the LPS-induced secretions of NO, TNF-α and IL-6. It also decreased the protein levels (IC₅₀?=?186?µg/mL) of LPS-induced inducible nitric oxide synthase (iNOS). The LPS stimulated p38, JNK and ERK phosphorylations significantly more than the controls. Surprisingly, although Ebn reduced p38 and JNK phosphorylations, it did not influence ERK phosphorylation. We found that Ebn revealed several major compounds such as chlorogenic acid (1.96?mg/100?g), rutin (46.48?mg/100?g), luteolin-7-glucoside (11.29?mg/100?g), naringin (1.13?mg/100?g), hesperidin (23.69?mg/100?g) and tangeretin (1.59?mg/100?g).

Discussion and conclusion: Boehmeria nivea exerts an anti-inflammatory effect on macrophages by inhibiting p38 and JNK, suggesting that it may be used as a functional ingredient against inflammation.     

Inhibitory potential of traditional herbs on α-amylase activity

Context: There has been enormous interest in the development of alternative medicines for the control of diabetes. Use of carbohydrate-hydrolyzing enzyme inhibitors proved to be an important strategy for the management of postprandial hyperglycemia by delaying the process of carbohydrate hydrolysis and absorption.

Objective: Three common traditional herbs, namely, stem bark of Terminalia arjuna (Combretaceae), seeds of Eugenia cumini (Myrtaceae), and leaves of Aegle marmelos (Rutaceae), were tested for their α-amylase inhibitory activities to establish antidiabetic potential.

Materials and methods: The plant extracts (aqueous, 50%, and 100% methanol) obtained were subjected to an in vitro amylase inhibitory assay using starch as a substrate and pancreatic amylase as the enzyme. Statistical differences and linear regression analysis were performed using GraphPad prism 5 software.

Results: The 50% methanol extracts of T. arjuna, E. cumini, and A. marmelos at a concentrations 50–500 μg/mL showed maximum percentage inhibition on amylase activity with IC₅₀ values of 302?±?0.55, 632?±?0.21, and 503?±?0.28 μg/mL, respectively. However, the 100% methanol extracts of all the three plants showed the least inhibitory activity.

Discussion and conclusion: The results show that T. arjuna > E. cumini > A. marmelos have excellent inhibitory activity and, therefore, might be effective in lowering postprandial hyperglycemia.     

Hepatoprotective effect of withanolide-rich fraction in acetaminophen-intoxicated rat: decisive role of TNF-α, IL-1β, COX-II and iNOS

Context: Overdose of acetaminophen (APAP) is common in humans and is often associated with hepatic damage. Withania somnifera (L.) Dunal (Solanaceae) shows multiple pharmacological activities including antioxidant and anti-inflammatory potential.

Objective: To evaluate the possible mechanism of hepatoprotective activity of withanolide-rich fraction (WRF) isolated from a methanolic extract of Withania somnifera roots.

Materials and methods: Hepatotoxicity was induced by oral administration of APAP (750?mg/kg, p.o.) for 14 d. The control group received the vehicle. APAP-treated animals were given either silymarin (25?mg/kg) or graded doses of WRF (50, 100 and 200mg/kg) 2?h prior to APAP administration. Animals were killed on 15th day and blood and liver tissue samples were collected for the further analysis.

Results: In WRF-treated group, there was significant and dose-dependent (p?<?0.01 and p?<?0.001) decrease in serum bilirubin, ALP, AST and ALT levels with significant and dose-dependent (p?<?0.01 and p?<?0.001) increase in hepatic SOD, GSH and total antioxidant capacity. The level of MDA and NO decreased significantly (p?<?0.01) by WRF treatment. Up-regulated mRNA expression of TNF-α, IL-1β, COX-II and iNOS was significantly down-regulated (p?<?0.001) by WRF. Histological alternations induced by APAP in liver were restored to near normality by WRF pretreatment.

Conclusion: WRF may exert its hepatoprotective action by alleviating inflammatory and oxido-nitrosative stress via inhibition of TNF-α, IL-1β, COX-II and iNOS.     

Hepatoprotective effects of Cassia semen ethanol extract on non-alcoholic fatty liver disease in experimental rat

Context: Cassia semen (Cs), a seed of Cassia obtusifolia L. (Leguminosae), is a popular functional beverage. Previous studies reported that Cs displayed antioxidant, antifungal and strong liver protective effects.

Objective: This study evaluates the hepatoprotective effects of Cs on non-alcoholic fatty liver disease (NAFLD).

Materials and methods: Seventy-two male Wistar rats raised with high-fat diet (HFD) were randomly allotted into model, metformin (0.2?g/kg) and Cs (0.5, 1, and 2?g/kg)-treated groups. Another 12 rats were raised with normal feed as control group; all the rats were orally administrated with drugs and vehicle for 6?weeks. Alanine transferase (ALT), aspartate transaminase (AST), triglycerides (TG), total cholesterol (TC), malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH), tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-8 and low density lipoprotein receptor (LDL-R) mRNA levels were measured at the end of the experiment.

Results: Twelve weeks of HFD administration significantly increased the levels of AST, ALT, TG, TC, TNF-α, IL-6, IL-8 and MDA, decreased SOD (199.42 vs. 137.70?U/mg protein) and GSH (9.76 vs. 4.55?mg/g protein) contents, compared to control group. Cs administration group significantly decreased the elevated biomarkers with the ED₅₀?=?1.2?g/kg for NAFLD rats. Cs treatment also prevents the decreased expression of LDL-R mRNA, and improved the histopathological changes compared to model group.

Conclusions: The hepatoprotective effect of Cs on NAFLD may possibly be due to its antioxidant effect. Cs may become a potent hepatoprotective agent in clinical therapy in the future.     

Immunomodulatory effect of Aegle marmelos leaf extract on freshwater fish Cyprinus carpio infected by bacterial pathogen Aeromonas hydrophila

Context: Aquatic organisms (especially fish) require potent defense mechanisms to protect themselves against pathogen invasion and disease formation. The use of immunostimulants in fish culture can prevent the diseases through augmentation of both specific and non-specific immunity.

Objective: A study was conducted to investigate the efficacy of different dietary doses of Aegle marmelos (Linn.) Corr. Serr. (Rutaceae) leaf extract for the immune response and the disease resistance of the freshwater fish, Cyprinus carpio Linn. (Cyprinidae) infected by Aeromonas hydrophila Chester (Aeromonadaceae).

Materials and Methodology: Hematological, specific immune response, non-specific immune response and enzyme assay studies were performed on fish and were scrutinized after 50 days of feeding trial.

Results: Fish were challenged with Aeromonas hydrophila at a dose of 1.5?×?10⁴ cells/mL through intraperitoneal injection, and the hematological changes, the immune response, the enzyme activity and the disease resistance of Cyprinus carpio against the pathogen were also studied for 20 days at 5-day intervals.

Discussion: The results obtained from the study demonstrated that the fish fed with leaf extract of Aegle marmelos incorporated into feed significantly enhanced the red blood cell count, white blood cell count, hemoglobin, phagocytic activity, nitroblue tetrazolium chloride assay, lysozyme, pathogen clearance and enzyme activity compared with the control group. The survivability was higher in the fish which consumed leaf extract-incorporated feed, and the fish group fed with 5?g diet showed highest percentage survival of the fish.

Conclusion: These results indicate that Aegle marmelos stimulates the immunity and makes the freshwater fish Cyprinus carpio more resistant to Aeromonas hydrophila.     

川芎嗪对N-甲基-N-亚硝基脲致光感受器细胞损伤的保护作用及其机制川芎嗪对N-甲基-N-亚硝基脲致光感受器细胞损伤的保护作用及其机制

目的探讨川芎嗪对N-甲基-N-亚硝基脲(N-methyl-N-nitrosourea，MNU)致大鼠视网膜损伤的作用。方法 大鼠生后47 d，腹腔注射不同剂量的川芎嗪注射液；50 d注射MNU 60 mg·kg^-1。测量视网膜总厚度；TUNEL和RT-PCR法分别检测细胞凋亡、c-jun和c-fos的表达。结果川芎嗪注射液可剂量依赖性地增加周边视网膜总厚度和降低凋亡指数，并抑制MNU诱导的即早基因表达。结论川芎嗪通过下调基因c-jun和c-fos的表达，抑制光感受器细胞凋亡，从而部分保护MNU引起的视网膜损伤。     

Hepatoprotective effect of Stachys pilifera ethanol extract in carbon tetrachloride-induce hepatotoxicity in rats

Context: Stachys pilifera Benth (Lamiaceae) has long been used to treat infectious diseases, respiratory and rheumatoid disorders in Iranian folk medicine. Antitumor and antioxidant activity of the plant have been reported.

Objective: The study was designed to assess the hepatoprotective activity of ethanol extract of Stachys pilifera in carbon tetrachloride (CCl₄)-induced hepatotoxicity in rats.

Materials and methods: The rats were randomly divided into six equal groups (n?=?7). Group I was treated with normal saline; Group II received CCl₄ (1?mL/kg. i.p., twice a week) for 60 consecutive days; Groups III, IV and V were given CCl₄ plus Stachys pilifera (100, 200 and 400?mg/kg/d,p.o.); Group VI received the extract (400?mg/kg/d, p.o.). Histopathological analysis and measurement of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), malondialdehyde (MDA), total protein (TP) and albumin (ALB) were performed.

Results: CCl₄ caused a significant increase in the serum levels of AST, ALT, ALP and MDA as well as decreased ALB, and TP serum levels (p?4-elevated levels of ALT, AST, ALP and MDA (p?4 group (p4.

Discussion: The results revealed that the Stachys pilifera extract could provide considerable protection against CCl₄ hepatotoxicity in rats that may be related to its antioxidant properties.