Liver Marker Enzymes Research Articles

PHARMACOLOGICAL ACTIVITY OF ULVA LACTUCA POLYPHENOLS FRACTION: HEPATOPROTECTIVE AND ANTIOXIDANT ACTIVITIES AGAINST PARACETAMOL-INDUCED LIVER DAMAGE IN RATS

Objective: The objective of this study was to assess the activity Ulva lactuca polyphenols fraction in protecting the liver damage induced by high dose of paracetamol.
 Methods: This study was performed using Wistar albino rats divided into six groups. Group 1 was the normal group. Groups 2, 3, 4, 5, and 6 received paracetamol (2 g/kg) for 7 days. In addition to paracetamol, Groups 3, 4, 5, and 6 received silymarin (100 mg/kg), U. lactuca polyphenols fraction at the doses of 50, 100, and 200 mg/kg, respectively, for 7 days. On the 8th day, serum and liver samples were collected from the animals and the hepatoprotective and antioxidant activities were assessed by studying the levels of liver marker enzymes, bilirubin, protein, reduced glutathione, and antioxidant enzymes.
 Results: U. lactuca polyphenols fraction, at the tested doses, restored the levels of all serum markers and enzymes (aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, lactate dehydrogenase, γ-glutamyl transferase, total bilirubin, total protein, cholesterol, triglycerides, and reduced glutathione) and liver homogenate markers (reduced glutathione, superoxide dismutase, catalase, and glutathione peroxidase) significantly, in dose-dependent manner.
 Conclusion: This study suggests that U. lactuca polyphenols fraction has a hepatoprotective effect against paracetamol-induced liver damage and possesses antioxidant activities.

MIMOSINE - A NOVEL SUBSTANCE AND EVALUATE THE LIPID PEROXIDATION

Objective: The mimosine is a novel substance extracted and purified from Mimosa pudica. The present study is to evaluate lipid peroxidation (LPO).
 Methods: The sox halation is used to differentiate extracts. Mimosine is purified by gas chromatography–mass spectrometry (GC–MS) and liquid chromatography–mass spectrometry (LC–MS). The present study was conducted to evaluate the hepatoprotective activity of ethanol extract of M. pudica against alcohol-induced albino rats. This was evident from significant of M. pudica on LPO and antioxidants liver of rats.
 Results: By GC–MS and LC–MS, the mimosine is purified and it shows the highest peak. The phytochemical analysis of M. pudica and the activity of the enzyme antioxidants such as superoxide dismutase is 176.83 g and peroxidase is 0.989 g, were found to be present in the plant extract of M. pudica. In rat treated with toxic dose of alcohol, a significant increase in serum and liver were observed. Simultaneous oral administration of the aqueous extract of M. pudica reduced the lactate dehydrogenase (LDH) activity to a significant extent compared to control. LDH, a cytosolic liver marker enzyme, is a regulator of many biochemical reactions in the body tissues and the fluid.
 Conclusion: The result obtained in our work, existence of bioactive substance proves that it is mainly used as a traditional medicine. The phytochemical analysis indicated the presence of alkaloids, steroids, terpenoids, tannin, phenolic, saponins, proteins, and amino acids. The liver showed that the induction of alcohol causes hepatic necrosis and injury, but it is cured by M. pudica.

Protective effect of atorvastatin on oxidative stress in streptozotocin-induced diabetic rats independently their lipid-lowering effects.

In the present study, we investigate the effects of atorvastatin on the lipid profile, oxidative stress, and liver enzyme markers, and its protective activity against diabetic complications, in streptozotocin (STZ)-induced diabetic rats. Fasting blood glucose (FBG), triglyceride (TG), total cholesterol (TC), and high-density lipoprotein (HDL) levels, as well as alanine aminotransferase (ALT) and aspartate aminotransferase (AST) enzyme activities, were measured 7 weeks after the administration of STZ and atorvastatin. Thiobarbituric acid reactive substances (TBARS), non-protein associated sulfhydryl (NP-SH), total sulfhydryl (T-SH), and nitric oxide (NO) levels were measured to evaluate oxidative stress. Atorvastatin was found to inhibit ALT and AST activities and to reduce FBG levels in rats with STZ-induced diabetes. Moreover, atorvastatin treatment significantly reduced lipid peroxidation in kidney, heart, and eye tissues (P < 0.001, for all), and resulted in a significant increase in NP-SH levels in brain tissues (P < 0.001). Total NO and nitrate levels increased significantly after atorvastatin treatment (P < 0.01). Our results revealed that atorvastatin has a protective effect against STZ-induced oxidative damage by reducing TBARS levels and increasing NP-SH levels, has a hepatoprotective effect by decreasing ALT and AST activities. It also shows the antihyperglycemic activity by lowering FBG levels.

Solanum nigrum confers protection against CCl4-induced experimental hepatotoxicity by increasing hepatic protein synthesis and regulation of energy metabolism

BackgroundHepatotoxicity induced by carbon tetrachloride (CCl4) is used as an experimental model to screen phytochemicals with liver protecting activity. Solanum nigrum (SN) is a shrub which is widely distributed throughout India. Traditionally the plant has been used for curing various ailments related to gastroenterology and hepatology. The present study was performed to evaluate the hepatoprotective property of S. nigrum extract standardized for its flavonoid content against CCl4-induced hepatotoxicity.MethodsThe liver protecting property of SN was evaluated by means of various biochemical parameters and histopathologically. S. nigrum was administered to Swiss strain female albino mice with either 100/ 200/ 300 mg/kg body weight/day for 30 days along with CCl4 which is a well established model to induce hepatotoxicity.ResultsAdministration of CCl4 for 30 days caused a significant increase in liver marker enzymes and a decrease in hepatic DNA, RNA and protein levels which was effectively mitigated by treatment with the plant extract in a dose dependent manner. Similarly co-treatment of the extract along with the hepatotoxin improved hepatic energy status by increasing the activities of succinate dehydrogenase (SDH) and adenosine triphosphatase (ATPase). Histopathological findings indicated severe vacuolization and necrotic changes after CCl4 treatment which was mitigated by the co-administration of SN extract in a dose-dependent manner.ConclusionsCarbon tetracholoride is known to exert hepatotoxicty by forming adducts with tissue macromolecules through covalent interactions. S. nigrum extract was found to effectively mitigate CCl4-induced changes in hepatic macromolecular content and energy status of liver tissue. The present study has identified the standardized S. nigrum as an effective hepatoprotective agent probably due to its role in improving the protein and energy levels in the hepatic tissue.

Effect of Ethanol Leaf Extract of Spilanthes filicaulis on Liver Indices and Selected Biochemicals on Carbon Tetrachloride Induced Hepatic Damage in Wistar Rats

This study investigated the effect of ethanol leaf extract of Spilanthes filicaulis on liver indices and selected biochemicals on carbon tetrachloride induced hepatic damage in wistar rats. A total of thirty (30) rats comprising of five (5) rats per group (90-130 g), were used for the study. Groups 1-3, served as normal, negative (CCl4 induced 1 mL/kg, 1:1 i.p.) and positive (CCl4 induced and administered with silymarin 50 mg/kg) controls respectively. Groups 4-6 were the extract treated groups administered with 250 mg/Kg bw, 500 mg/Kg bw and 750 mg/Kg bw respectively for 14 days. The results of the liver marker enzymes showed a significant decrease (p 0.05) in all treated groups when compared to the negative control group. Catalase (CAT) and Superoxide Dismutase (SOD) levels were significantly increased (p 0.05) in all treated groups when compared to the negative control. The study showed that the ethanol leaf extract of Spilanthes filicaulis has potential of restoring hepatic liver damage.

Nutritive properties of aqueous extract Justicia carnea leaves and its effects on haematological and some biochemical indices of anaemia induced male wistar albino rats

This study investigated nutritive properties of aqueous extract of J. carnea leaves and its effects on haematological and biochemical indices of anaemia induced male Wistar albino rats. In the nutritional study, proximate composition, phytochemicals, antinutritonal factors, minerals and vitamins analyses were carried out using standard analytical procedures. Eighteen male Wistar albino mice were used for the acute toxicity study while 28 male Wistar albino rats were used for haematological and biochemical study. The rats were randomly distributed into seven groups of four rats each with group 1 as normal control. Group 2 was anaemia induced untreated, and group 3 was anaemia induced treated with 50 mg/kg b. wt. of Astyfer. Groups 4-6 were anaemia induced and treated with 100, 300 and 500 mg/kg b. wt. of the extract respectively. Group 7 was non-anaemic rats that received 500 mg/kg b. wt. of the aqueous extract. Nutritional study showed richness of the aqueous extract in proximate composition, phytochemicals, minerals and vitamins contents. The extract caused no adverse reactions or death in the animals indicating it is relatively safe for consumption. Significant (p<0.05) reduction in the levels of haematological indices observed in anaemic rats were significantly (p<0.05) reverted within 4 to 8 d of treatment with the extract when compared with group 2. There were no significant difference observed in the liver marker enzymes activities and renal function indices concentrations of all the anaemic rats treated with the aqueous extract when compared with normal control, which could be attributed to the aqueous extract lacking potentials of causing liver and renal damage. These findings suggest that aqueous of J. carnea leaves is safe, nutritionally rich with no toxic effects on liver and kidney function indices, and possesses anti-anaemic effects due to its blood boosting and replenishing properties.

Hepatoprotective properties of methanol leaf extract of Pterocarpus mildbraedii Harms on carbon tetrachloride- induced hepatotoxicity in albino rats (Rattus norvegicus)

Aim: This study evaluated the effects of methanol leaf extract of Pterocarpus mildbraedii Harms on carbon tetrachloride (CCl4)-induced sub-acute hepatotoxicity in albino rats. Methods: Fresh leaves of P. mildbraedii were collected in December 2016. Thirty male albino rats were randomly assigned to 6 groups (A – F) of 5 rats each. Sub-acute hepatotoxicity was induced in groups A, B, C, D and E by intraperitoneal injection of 1ml/kg CCl4 in equal volume of olive oil at 3-day intervals for 12 days. Group A was given 10 ml/kg distilled water placebo and served as untreated (negative) control, groups B, C, and D were treated with 100, 200 and 400 mg/kg P. mildbraedii methanol leaf extract (PME) respectively, group E was treated with 100 mg/kg silymarin as positive control, while group F was given 10 ml/kg distilled water placebo and served as normal control. Treatment with PME and silymarin was done orally twice daily for 15 days. Blood samples were collected on the 15th day for evaluation of liver enzyme markers (alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) activities) and liver function (total serum protein, albumin, globulin, total cholesterol and bilirubin), following standard procedures. Relative liver weight was calculated. Results: Treatment with PME and silymarin significantly (P < 0.05) decreased the elevated ALT and AST, and thus restored hepatocellular integrity, and also ameliorated inflammatory liver enlargement in the rats given CCl4. Conclusion: These findings imply that treatment with PME as used in this study led to significant hepatoprotection against CCl4-induced hepatotoxicity.

Evaluation of the phytochemical content, in vitro antioxidant capacity, biochemical and histological effects of Dennettia tripetala fruits in healthy rats.

We estimated the content of specific phytochemicals and in vitro antioxidant properties of the powder, aqueous, and ethanolic extracts of ripe Dennettia tripetala fruits. We also tested the biochemical and histological effects of these fruit extracts on healthy rats. Aqueous and ethanolic extracts were prepared from the powder of ripe D. tripetala fruits, and standard phytochemical methods were used to evaluate its phytochemical content and antioxidant properties. Eighteen rats were randomized into three groups, one of which served as control, while the second and third groups received the aqueous and ethanolic extracts of D. tripetala fruits, respectively, at a dose of 1,000 mg/kg bw daily for 28 days. Our results show that the powder as well as the aqueous and ethanolic extracts of ripe D. tripetala fruits contains phenols, flavonoids, saponins, tannins, and alkaloids. The plant powder as well as both extracts scavenged DPPH and hydrogen peroxide as well as reduced ferric ions. The extracts of D. tripetala fruits did not alter liver marker enzymes or serum protein profile of the rats. The extracts also did not alter the serum concentration of urea and creatinine and the antioxidant enzyme activity and lipid peroxidation levels in the liver but altered that of the kidney. The extracts altered the serum and liver lipid profile but not to any significant extents. Also, the extracts caused minimal congestion to the centrioles of the liver but were not in any other way toxic to the liver, kidney, or heart of the rats. Our results point to the fact that the fruits of D. tripetala possess phytochemicals with medicinal properties and are well tolerated by rats.

Antioxidant traits and protective impact of Moringa oleifera leaf extract against diclofenac sodium-induced liver toxicity in rats.

Moringa oleifera gained importance as a medicinal plant. The current study assesses Moringa leaf ethanol extracts (MLE) against experimentally diclofenac sodium (DcNa)-induced liver toxicity in male rats. Leaves were extracted with different solvents differing in polarity. Assessment involved total phenolic compounds, total flavonoids and radical scavenging activity against 1,1-diphenyl-2-picrylhydrazyl (DPPH·). HPLC was performed for identifying phenolic compounds, wherein ethyl vanillin (1,205mg/kg), 3-OH-tyrosol (812.2mg/kg), benzoic acid (273.8mg/kg), salicylic acid (240.0mg/kg), chlorogenic acid (233.3mg/kg) and 3,4,5-methoxy-cinnamic acid (172.5mg/kg) were measured. Fifty animals (each treatment group consisted of 10 rats) were subjected to five treatments and the experiment lasted for 4weeks. Animals were exposed to DcNa (100mg/kg) and two doses of MLE as well as silymarin (an antioxidant flavonoid C25 H22 O10 ) for 4weeks. Liver marker enzymes, including alkaline phosphatase, alanine transaminase, and aspartate transaminase as well as urea, uric acid, and creatinine were increased. Serum albumin and total protein decreased in DcNa-treated rats. Homogenates nitric oxide increased in liver tissue of the DcNa-treated rats, while the activity of each of glutathione peroxidase, glutathione-S-transferase, glutathione, and catalase decreased. It could be concluded that MLE in both doses and silymarin are considerably hepatoprotective with antioxidant activity (AOA) against DcNa-induced hepatotoxicity in rats. PRACTICAL APPLICATIONS: Administration of MLE caused improvements in kidney functions and acted as antioxidant enzymes as compared with silymarin (as a reference drug). AOA was exhibited by MLE in vivo, and this would have a positive effect against oxidative liver damage caused by DcNa. Plasma membrane was protected and the regenerative and reparative capacity of liver increased by phenolics in the MLE. The study demonstrated the MLE hepatoprotective activity and recommends using M. oleifera leaves for the treatment of liver disorders.

English

Eighty male albino rats were used to investigate the effects of methanolic calyx extract of Hibiscus sabdariffa on body weight, blood cholesterol and liver enzymes markers. Twenty-eight days oral administration of 100, 200, 400 and 800 mg/kg body weight of the extract showed a significant (p 0.05) decreased the serum cholesterol and increased the liver marker enzymes (ALP, ALT and AST) in dose-dependent and time-dependent manner, when compared with the control. However, on day 21, the group treated with 400 mg/kg showed a significant (p<0.05) increase the serum cholesterol, and decrease in liver marker enzymes when compared with the rest of the treatment groups. Histopathology from all the treatment groups revealed graded degrees of vacuolar degeneration of the hepatocytes and peri-portal infiltration of mononuclear leucocytes. The results of this present study suggests that the methanolic calyx extract of H. sabdariffa possesses anti-obesity and hypocholesterolemic potentials which should be harnessed with caution due to its tendency to adversely affect the liver. Key words: Hibiscus sabdariffa, hypocholesterolemia, anti-obesity, liver maker enzymes, Wistar rats.

Hepato‐protective effects of R‐phycoerythrin‐rich protein extract ofPortieria hornemannii(Lyngbye) Silva against DEN‐induced hepatocellular carcinoma

The present investigation evaluates the hepato‐protective potential of R‐phycoerythrin (R‐PE)‐rich protein extract obtained from Portieria hornemannii (Lyngbye) Silva against H₂O₂‐induced hepatotoxicity using HepG2 cells (in vitro) and male Wistar albino Hepatocellular carcinoma (HCC) rats (in vivo). H₂O₂ treatment led to dose‐dependent decreases in cell viability of HepG2, which was ameliorated by the treatment with R‐PE‐rich extract. In vivo studies showed that all the vital parameters including enzymatic and nonenzymatic antioxidants as well as liver marker enzymes were affected in the experimentally induced HCC male Wistar rats upon exposure to the N‐diethylnitrosamine (DEN), a carcinogen. However, there was moderate to better restoration of such parameters in the carcinoma rats treated with R‐PE‐rich protein extract. PRACTICAL APPLICATIONS: Therapeutic alternatives originating from food or food supplements are gaining popularity as “nutritional therapy” and they are well studied for their chemo‐preventive effects. Investigations of several food‐derived bioactive compounds revealed their ability to antagonize dysregulated targets in cellular signaling pathways to exhibit antineoplastic activities. Isolation of bioactive molecules present in marine food products and determination of their broad range pharmaceutical activity by means of deducing specific molecular targets as well as establishing minimal toxicity to normal tissues could aid in treatment of cancer. Phycobiliproteins are an important group of pigment molecules extracted from the red and blue green algae. A study was envisaged to evaluate the anticancer potential of RPERPE (R‐PE‐rich protein extract) obtained from P. hornemannii (Lyngbye) Silva against HCC using male Wistar albino rats as models. This study offers the scope of using the R‐PE‐rich protein extract of P.hornemannii for developing therapeutic anticancer formulation.

Ameliorative effect of silymarin against linezolid-induced hepatotoxicity in methicillin-resistant Staphylococcus aureus (MRSA) infected Wistar rats.

Linezolid has a better choice for eradication of methicillin-resistant Staphylococcus aureus (MRSA) infections, but its use is limited because of linezolid-induced hepatotoxicity, myelosuppression, and lactic acidosis. This research elucidated the role of silymarin against hepatoxicity of linezolid therapy in MRSA infected Wistar rats. The rats were rendered neutropenic by an intraperitoneal injection of cyclophosphamide injection. The neutropenic rats were injected subcutaneously with 106 CFU/ml of MRSA. The rats were divided into 6 groups. Normal control, Infected, Infected animals treated with linezolid 50 mg/kg/twice/day and Infected animals treated with linezolid and different dose of silymarin 25, 50, and 100 mg/kg/twice/day for 14 days. On the 15th day, the blood, liver, kidney, and bone marrow were collected for biochemical and histopathological examination. The MRSA was confirmed by PCR assay. The minimum inhibitory concentration of linezolid was 0.5-2 μg/ml. The linezolid induced liver damage was confirmed by elevation of marker enzymes alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), Lactate dehydrogenase (LDH) levels, serum bilirubin, lactate, and histopathological studies of the liver. The linezolid treated rats also showed myelosuppression, lactic acidosis, oxidative stress and decreased intestinal alkaline phosphatase (IAP). The silymarin administration exhibited marked hepatoprotective effect by significantly lowering the liver marker enzymes, serum parameters, and cytological findings reflect the hepatoprotection. Additionally, Silymarin showed protection against myelosuppression and lactic acidosis evidenced by bone marrow smear and serum lactate estimation. Antioxidant effect of silymarin was confirmed by decreased levels of lipid peroxidation, restored the enzymatic and non-enzymatic antioxidants of the liver nearer to normal. The present study indicates that the silymarin could be a better herbal therapeutic agent which protects against the linezolid induced hepatotoxicity in MRSA infected rats.

In vivo toxicity evaluation of biologically synthesized silver nanoparticles and gold nanoparticles on adult zebrafish: a comparative study.

In this study, toxicity of biologically synthesized silver nanoparticles (AgNPs) and gold nanoparticles (AuNPs) was compared using zebrafish as a model organism. At 96h, LC50 of AgNPs and AuNPs was found to be 24.5µg/L and 41mg/L, respectively. Following the LC50 determination, half of the LC50 of AgNPs (12.25µg/L) and AuNPs (20.5mg/L) was exposed to adult zebrafishes for 14days. Morphological changes, liver marker enzymes, reactive oxygen species (ROS) generation, genotoxic effects and mRNA expression levels of oxidative stress and innate immune response related genes were studied using nanoparticle treated gill, liver and blood cells. In this study, AgNP-treated gill and liver tissues showed a number of morphological changes such as cell membrane damage, irregular cell outlines, pyknotic nuclei and complete disruption of gill and liver cells; on the contrary, AuNPs treated liver tissues alone showed such changes. The levels of liver marker enzymes such as alanine aminotransferase and aspartate aminotransferase were increased after AgNPs treatment when compared to AuNPs treatment. AgNP-treated liver cells showed higher levels of ROS generation than the control; on the other hand, AuNPs treatment exhibited lower levels of ROS generation than the control. Interestingly, AgNP-treated blood cells showed micronuclei formation and nuclear abnormalities, while AuNPs treatment did not show such effects. Based on these observations, it is clear that AgNPs may cause oxidative stress and immunotoxicity to adult zebrafish than the AuNPs. However, these results clearly reveal the significance of relatively safe and less toxic bionanomaterials for possible biomedical applications.

Modulation of Oxidative Stress and Hyperglycemia by Rheum spiciformis in Alloxan Induced Diabetic Rats and Characterization of Isolated Compound.

This study evaluates the ameliorative potential of Rheum spiciformis methanolic (RS-MeOH) extract in reducing oxidative stress and hyperglycemia in albino rats along with characterization of possible therapeutic compound(s). Groups treated with 50 and 100 mg/kg bw plant extract (RS-MeOH ) decrease blood glucose levels from 359.9±8.2 to 209.5±8.5 mg/dl (50 mg/kg bw) and 354.7±13.3 to 162.5±7.4 mg/dl (100 mg/kg bw) on the 0th and 14th day (P<0.001) respectively. This reduction in blood glucose was significant as compared to glibenclamide (20 mg/dl) which reduced glucose levels from 297.7±11.39 to 132.9±8.74 mg/dl on 0th and 14th day respectively. Biochemical parameters triglycerdies, cholesterol, low density lipoprotein (LDL) and creatinine were also reduced in a dose dependent manner. Liver marker enzymes were positively modulated by administration of RS-MeOH (P<0.001). Antioxidant enzyme profile showed an enhanced/better pattern after the administration of RS-MeOH extracts for reduced glutathione, reduced glutathione (GR), glutathione peroxidase (GPx), glutathione-S-transferase (GST) and superoxide dismutase (SOD) in both liver and pancreas. Moreover pancreatic histopathology reports revealed β-cell restorative effects with RS-MeOH, thereby potentiating its role in improving blood glucose levels. RS-MeOH purification and isolation studies involving GC-MS and NMR techniques revealed presence of emodin type compounds in RS-MeOH. Overall Rheum spiciformis showed ameliorative action on oxidative stress and hyperglycemia, however further studies to explore the mechanism of action of possible therapeutic compound in invivo clinical trials will prove beneficial for the advancement of new oral antidiabetic drug.

Premna integrifolia ameliorates cyclophosphamide-induced hepatotoxicity by modulation of oxidative stress and apoptosis

The present study was designed to evaluate the ameliorative effect of ethyl acetate extract of Premna integrifolia L. (EAEPI) leaves in cyclophosphamide (CP)-induced hepatic injury in mice. Mice were intoxicated with CP (200 mg/kg b. wt., i.p.) for 5 weeks or EAEPI (400 and 600 mg/kg b. wt., orally) in combination with CP. The results demonstrated that EAEPI exerts protective effect against CP induced hepatotoxicity, as evident from restoration of altered haematological parameters and alleviations of liver marker enzymes in serum. EAEPI also attenuated oxidative stress and antioxidant markers as evident from reversal of lipid peroxidation, glutathione levels as well as activities of catalase and superoxide dismutase enzymes. Moreover, EAEPI attenuated apoptosis and histopathological liver tissue damage in CP-intoxicated mice. In conclusion, EAEPI could protect mice liver against cyclophosphamide toxicity by inhibiting oxidative stress and apoptosis.The protective activity of EAEPI may be due to presence of polyphenolic compounds as identified by UHPLC–Q-TOF-MS/MS.

IN VITRO ANTIOXIDANT POTENTIAL AND HEPATOPROTECTIVE ACTIVITY OF TAXUS WALLICHIANA

Objectives: Taxus wallichiana, the member of family Taxaceae is used for the cure of various ailments by a large ethnic population native to the forests in Kashmir valley. The aim of this study is to evaluate antioxidant potential and hepatoprotective action of T. wallichiana as claimed by the ethnic population segment. Methods: Antioxidant potential of different T. wallichiana extracts was evaluated by estimating the total phenolics, 1,1-diphenyl -2-picrylhydrazyl (DPPH), reducing power, microsomal lipid peroxidation (LPO), and hydroxyl radical scavenging activity through standard protocols. Hepatoprotective activity of T. wallichiana was determined by evaluation of liver marker enzyme (aspartate transaminase, alanine transaminase, and alkaline phosphatase) levels using standard protocols and by histological analysis of rat liver tissues of different animal groups against CCl4 induced liver damage. Results: Phenolic content expressed as gallic acid equivalents (GAE/g) was found maximum in case of ethanolic extract (153.34 mg GAE/g), followed by extracts of methanol (149.24 mg GAE/g), hexane (134 mg GAE/g), ethyl acetate (128.9 mg GAE/g), chloroform (121.37 mg GAE/g), and aqueous (113.00 mg GAE/g). At concentration of 700 μg/ml, DPPH radical scavenging activity of methanol extract was 88.29% with IC50 (212.00 μg/ml), aqueous (82.47%) with IC50 (258.29 μg/ml), and ethyl acetate (79.57%) with IC50 (301.80%μg/ml). The reducing power of the extracts increased in a concentration dependent manner. At concentration of 700 μg/ml, 79%, 68%, and 56% inhibition was observed with extracts of methanol, aqueous, and ethyl acetate on microsomal LPO with IC50 values of 126.09, 168.83, and151.96 μg/ml, respectively. Superoxide radical scavenging activity of T. wallichiana extracts increased in a dose-dependent manner with IC50 values 170.30 μg/ml (methanol), 257.00 μg/ml (aqueous), and 297.55 μg/ml ethyl acetate extract. T. wallichiana extracts exhibited antioxidant effects on calf thymus DNA damage. The results further depict the hepatoprotective action of T. wallichiana extract at doses of 100 and 300 mg/kg and were comparable with that of standard treatment comprising 50 mg/kg-day of Vitamin C, a known hepatoprotective agent. The data were substantiated with histopathological studies of rat liver sections. Conclusion: These results comprehensively depict that T. wallichiana extracts are endowed with the free radical sequestering potential and be employed as potential antioxidant and hepatoprotective arsenal against many oxidative stress linked diseases.

KADMİYUMA MARUZ BIRAKILAN DİYABETİK RATLARA ALFA LİPOİK ASİT VE İNSÜLİN UYGULANMASININ OLASI DÜZENLEYİCİ ETKİLERİ

OBJECTIVE: Environmental exposure to the cadmium (Cd), is associated with hyperglycemia and reduced serum insulin. This investigation was planned to assess the effects of Lipoic Acid (LA) and insulin on glycolytic enzymes, liver marker enzymes and lipids in Cd exposed diabetic rats. MATERIAL AND METHODS: Male Wistar rats were separated into 7 groups (n=8 in each group). Groups were designed as control, diabetic control, diabetic + CdCl2, diabetic + insulin, diabetic + CdCl2 + insulin, diabetic + CdCl2 + LA, anddiabetic + CdCl2 + insulin + LA groups. Type 1 diabetes was established by intraperitoneal (i.p.) injection of streptozotocin (STZ) (65 mg/kg) into 6 groups. Insulin (4 IU/kg/day) was given subcutaneously (s.c.) to insulin treated groups. CdCl2 (1,2 mg/kg/day) was given s.c. to CdCl2 treated groups. LA (100 mg/kg/day) was given i.p. to LA treated groups. CdCl2, LA, and insulin treatment were started 2 days after intraperitoneal STZ injection and continued for 3 weeks. Serum glucose, AST, ALT, BUN, LDL, HDL, and TG levels and liver hexokinase (HK), pyruvate kinase (PK), whole blood HbA1c level, and Na+/K+ATPase activity were evaluated. RESULTS: In diabetic group, serum glucose, HbA1c, TG, LDL, AST, ALT, ALP, and BUN levels were higher than cont- rol, but HDL was lower. In liver tissue, activities of Na+/ K+ATPase, HK and PK activities were decreased in dia- betic control group. PK, HK and Na+/K+ATPase activities were increased in liver in diabetic+CdCl2 and Diabeti- c+Insulin+CdCl2 groups. An increase was determined in activities of HK, PK, and Na+/K+ATPase in insulin and LA treated groups compared with diabetic control group. CONCLUSIONS: These results suggest that application of insulin and LA could be an effective therapeutic intervention against liver injury caused by Cd and STZ.

Rutin protects t‑butyl hydroperoxide-induced oxidative impairment via modulating the Nrf2 and iNOS activity

BackgroundRutin (quercetin-3-O-rutinoside), a flavonoid, is predominantly found in the buckwheat, cranberries, mulberry and citrus fruits. It is used as a restorative in the preparation of herbal medicine, multivitamin and known to reduce the fate of heart attack. HypothesisWe aimed to elucidate whether rutin attenuates oxidative stress and its possible mechanism of action in ameliorating the deleterious effect of t-BHP. We also provide evidence that rutin protects the antioxidant status of erythrocytes and liver via Nrf2 and iNOS pathway from oxidative stress. Study Design/MethodHuman erythrocytes and mice liver were used for the evaluation of rutin's effect against t-BHP induced oxidative stress. The non-enzymatic (GSH, MDA, -CO, –SH) and enzymatic stress markers (SOD, CAT, GPx, GR and GST) were estimated by the colorimetric method. The level of Nrf2, iNOS, liver marker enzymes, triglycerides, cholesterol, HDL-cholesterol, albumin, BUN was measured using ELISA kits. Reactive oxygen species (ROS) was quantified using flow cytometry and fluorometry. RT-PCR was used for the quantification of Nrf2 and iNOS expression levels in the liver tissue of mice. In silico studies were done through receptor-ligand binding interaction. ResultsPre-treatment with the rutin ameliorated the toxic effect of t-BHP by modulating the basal level of GSH, -SH, MDA and –CO significantly (p < 0.01) with respect to untreated control. Rutin also protected the erythrocytes against the t-BHP-induced oxidative stress as evidenced by augmented activity of antioxidant enzymes (CAT, SOD, GPX, GR and GST). Furthermore, at the highest tested concentration (16.3 µM), it protected the morphology of the erythrocytes by decreasing the ROS level (p < 0.01). In addition, the lower MEF values of rutin (0.520 ± 0.005) alone or along with t-BHP (0.630 ± 0.021) indicated its non-toxic and protective behavior. The qPCR analyses revealed that t-BHP potently up-regulates the iNOS and down regulate the Nrf2 expression which was ameliorated with rutin treatment in a dose-dependent manner like silymarin. ConclusionOur findings demonstrate that rutin potentiates its beneficial aspect by displaying a profound role in iNOS-Nrf2 signaling pathway. Accordingly, it may be concluded that the dietary factors wherein rutin is an ingredient could be helpful in the maintenance of the intracellular redox-homeostasis and thus may be effective against oxidative stress related secondary complications.

Antineoplastic Approach of Semecarpus anacardium Leaves Against N-Nitroso Diethylamine Initiated Hepatocellular Carcinoma

Abstract: Objective: To evaluate the effect of MESA on NDEA induced hepatocellular carcinoma on Sprague Dawley rats. Methodology: Semecarpus anacardium commonly known as ‘marking nut tree’ is a rich antioxidant and in Ayurveda it has been used for treatment of various forms of cancer. Methanolic extract of leaves of Semecarpus anacardium (MESA) was studied for its potential antioxidant property both in vitro and in vivo. NDEA 200 mg/kg, single i.p was the hepatocarcinogen while 200 mg/kg and 400 mg/kg of MESA was administered daily orally for 12 weeks. Protective effect of MESA was evaluated by measuring biochemical parameters like SGOT, SGPT, ALP and LDH. Alpha fetoprotein test was performed and histopathology changes of livers were assessed. Results: Preliminary phytochemical study revealed the presence of flavonoids, tannins, steroids, alkaloids, glycosides and vitamin C. MESA restored the level of antioxidants near to normal and reduced the elevated serum level of SGOT, SGPT, AST, LDH and AFP. The histopathology changes i.e. necrosis, widened sinusoids, elevated inflammatory cell infiltrate were partly or fully prevented in animals treated with the extract. Conclusion: The result of the present study indicates MESA to be rich in antioxidants and its active constituents mediated the free radical scavenging activity. 400 mg/kg extract showed promising effect in management of primary liver cancer. Key words: Semecarpus anacardium, Hepatocellular carcinoma, N-nitroso diethylamine, Alpha fetoprotein, Liver marker enzymes.

COMPARATIVE STUDY OF BUTANOLIC AND ETHANOLIC EXTRACTS OF TRIBULUS TERRESTRIS FRUITS IN 2,3,7,8-TETRACHLORODIBENZO-P-DIOXIN-INDUCED HEPATOTOXICITY IN RATS

Objective: To compare hepatoprotective activity of ethanol and butanol fraction of Tribulus terrestris fruits against 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-induced hepatic damage in albino rats.Methods: The degree of liver protection was assessed in terms of biochemical parameters, liver marker enzymes, and lipid peroxidation (LPO) in drug-treated and TCDD-intoxicated rats.Results: The results revealed that the elevated serum marker enzymes and biochemical parameters in TCDD-treated rats were significantly reduced toward normal levels in saponin rich fraction of T. terrestris drug-treated animals when comparable to ethanolic extract of T. terrestris drug-treated group. The plant extracts have got a notable inhibitory activity on the formation of LPO products in the basal as well as in the presence of inducers.Conclusion: The plant extracts hindered oxidative damage that occurred to the hepatocytes during TCDD toxicity may be responsible for the hepatoprotective activity in the present study.