Rat Hepatic Tissue Research Articles

Chemical composition, antioxidant and macromolecule damage protective effects of Picrorhiza kurroa Royle ex Benth

In the present study, we identified the chemical constituents of 70% hydroalcoholic fraction of Picrorhiza kurroa by LC–ESI–MS/MS which showed the presence of iridoid glucosides such as picroside I, picroside II, picroside III, picroside IV, kutkoside, pikuroside and flavonoids like apocynin and vanillic acid. P. kurroa exhibited DPPH radical scavenging and metal chelating activities with IC50 of 75.16±3.2 and 55.5±4.8μg/mL and also showed potent reducing power and total antioxidant activities. The extract inhibited macromolecule damage such as H2O2 induced plasmid DNA damage and AAPH induced oxidation of bovine serum albumin and lipid peroxidation of rat hepatic tissues.

Ameliorating role of rutin on oxidative stress induced by iron overload in hepatic tissue of rats.

Iron is an essential element that participates in several metabolic activities of cells; however, excess iron is a major cause of iron-induced oxidative stress and several human diseases. Natural flavonoids, as rutin, are well-known antioxidants and could be efficient protective agents. Therefore, the present study was undertaken to evaluate the protective influence of rutin supplementation to improve rat antioxidant systems against IOL-induced hepatic oxidative stress. Sixty male albino rats were randomly divided to three equal groups. The first group, the control, the second group, iron overload group, the third group was used as iron overload+rutin group. Rats received six doses of ferric hydroxide polymaltose (100 mg kg(-1) b.wt.) as one dose every two days, by intraperitoneal injections (IP) and administrated rutin (50 mg kg(-1) b.wt.) as one daily oral dose until the sacrificed day. Blood samples for serum separation and liver tissue specimens were collected three times, after three, four and five weeks from the onset of the experiment. Serum iron profiles total iron, Total Iron Binding Capacity (TIBC), Unsaturated Iron Binding Capacity (UIBC), transferrin (Tf) and Transferrin Saturation% (TS%)}, ferritin, albumin, total Protein, total cholesterol, triacylglycerols levels and aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities were determined. Moreover, total iron in the liver, L-malondialdehyde (L-MDA), glutathione (GSH), Nitric Oxide (NO) and Total Nucleic Acid (TNA) levels and glutathione peroxidase (GPx), catalase (CAT) and superoxide dismutase (SOD) activities were also determined. The obtained results revealed that, iron overload (IOL) resulted in significant increase in serum iron, TIBC, Tf, TS% and ferritin levels and AST and ALT activities and also increased liver iron, L-MDA and NO levels. Meanwhile, it decreased serum UIBC, total cholesterol, triacylglycerols, albumin, total protein and liver GSH, TNA levels and Gpx, CAT and SOD activities when compared with the control group. Rutin administration to iron-overloaded rats resulted in significant decrease in serum total iron, TIBC, Tf, TS%, ferritin levels and AST and ALT activities and liver total iron, L-MDA and NO levels with significant increases in serum UIBC, albumin, total protein and total cholesterol levels and in liver GSH, CAT and SOD activities compared with the IOL group. This study provides in vivo evidence that rutin administration can improve the antioxidant defense systems against IOL-induced hepatic oxidative stress in rats. This protective effect in liver of iron-loaded rats may be due to both antioxidant and metal chelation activities.

The Influence of Collagen III Expression on the Efficiency of Cell Isolation With the Use of Collagenase H

ObjectiveWe previously demonstrated that collagenase H (ColH) plays a crucial role in rat islet isolation, whereas collagenase G (ColG) plays only a supporting role. We also showed that collagen III appears to be one of the key targets of ColH based on a mass spectrometry analysis. In the present study, we investigated whether our novel findings in an islet isolation model are universally applicable for other types of cell isolation, such as a hepatocyte isolation, with the use of enzyme blends of recombinant collagenases. MethodsAs the first step, the expression of one of the main matrix components, collagen III, on rat pancreatic and hepatic tissues was assessed with the use of immunohistochemical staining. ColG and ColH were expressed in recombinant E. coli carrying expression plasmids for each collagenase. Then the efficiency of the collagenase subtype on rat hepatocyte isolation was evaluated in terms of cell yield with the use of thermolysin combined with either ColG or ColH (n = 3, respectively). ResultsThe expression of collagen III on rat hepatic tissues was dramatically lower than that of rat pancreatic tissues. In the rat hepatocyte isolation, a substantial amount of hepatocytes (0.81 ± 0.11 × 106) were obtained in the ColG group, whereas almost no hepatocytes were retrieved in the ColH group, indicating that the influence of the collagenase subtypes in rat hepatocyte isolation are completely opposite to that observed in rat islet isolation. ConclusionsConsidering that the expression of collagen III on hepatic tissues was relatively low and that almost no hepatocytes were retrieved when ColH and thermolysin were used, the present study supports our novel finding that collagen III appears to be one of the key targets of ColH in hepatocyte isolation. Therefore, the semiquantification of collagen III on the target tissues not only may positively contribute to efficient islet isolation, but also may affect other types of cell isolation by optimizing the ColH amount.

Effect of intrauterine growth retardation on gluconeogenic enzymes in rat liver

To investigate the expression of gluconeogenic enzymes phosphoenolpyruvate carboxykinase (PEPCK) and G-6-Pase mRNA of hepatic tissue in rats with intrauterine growth retardation (IUGR) and to explore the molecular mechanism of insulin resistance in IUGR rats. Pregnant rats were randomly divided into 2 groups: a normal group and a model group. The normal group were fed with 21% protein forage and the model group with 10% low protein forage to obtain IUGR pup rats. The pup rats were introduced to the normal group and the IUGR group prospectively. At 1, 3 and 8 weeks, the body weight, blood glucose, insulin concentration and insulin resistance index of the pup rats were measured. Expression of PEPCK and G-6-Pase mRNA were detected by RT-PCR. The birth weight of the IUGR group was significantly lower than that of the normal group (P<0.001). The weight of the IUGR group was still lower than that of the normal group at 1, 3 and 8 weeks. There was no significant difference in the blood glucose, insulin level and insulin resistance index between the 2 groups (P>0.05). The hepatic expression of PEPCK and G-6-Pase mRNA in the IUGR group was significantly higher than that of the normal group at 1, 3 and 8 weeks (P<0.01). The significantly increased expression of PEPCK and G-6-Pase mRNA of hepatic tissue in IUGR rats may increase gluconeogenesis, which is probably one of the molecular mechanisms of insulin resistance and diabetes in IUGR.

Iridoid Glycoside from the Leaves of Clerodendrum volubile Beauv. Shows Potent Antioxidant Activity Against Oxidative Stress in Rat Brain and Hepatic Tissues

ABSTRACTAim: This study aims at reporting the isolation, structure elucidation, and antioxidant potentials of ajugoside from C. volubile leaves in sodium nitroprusside (SNP)-induced oxidative stressed rat brain and hepatic tissues. Materials and Method: An iridoid monoterpene, ajugoside was isolated from the n-butanol fraction of C. volubile and evaluated for its antioxidant protective potential on brain and liver tissues of male Wister rats in an ex vivo model. Two molar concentrations (6.4 × 10−4 M and 1.28 × 10−3 M) of the metabolite and SNP were incubated with the tissues homogenate at 37°C for 2 hr prior to the test and assayed for catalase, superoxide dismutase (SOD) activities, and lipid peroxidation. α tocopherol (6.4 × 10−4 M) was used as standard. Results: Both molar concentrations exhibited high catalase activity in the tissues. However, 6.4 × 10−4 M ajugoside exhibited a very high SOD activity (liver: 96.45 and brain: 96.30%) and inhibition of lipid peroxidation (liver: 88.11 and brain: 93.27%) compared to the standard. 1.28 × 10−3 M ajugoside also exhibited good activities but lower than that of the standard and 6.4 × 10−4 M ajugoside. Discussion and Conclusion: Ajugoside showed potent antioxidant activities as evidenced by the synergistic high activities of SOD and catalase as well as inhibition of lipid peroxidation in the studied tissues.

Protective effects of pine bark extract against cisplatin-induced hepatotoxicity and oxidative stress in rats.

We investigated the protective effects of pine bark extract (pycnogenol®, PYC) against cisplatin-induced hepatotoxicity and oxidative stress in rats. Twenty-four male rats were divided into the following four groups: (1) vehicle control, (2) cisplatin (7.5 mg/kg), (3) cisplatin & PYC 10 (10 mg/kg/day), and (4) cisplatin & PYC 20 (20 mg/kg/day). A single intraperitoneal injection of cisplatin induced hepatotoxicity, as evidenced by an increase in serum aminotransferase and histopathological alterations, including degeneration/necrosis of hepatocytes, vacuolation, and sinusoidal dilation. In addition, an increase in the malondialdehyde (MDA) concentration and a decrease in the reduced glutathione (GSH) content and catalase (CAT), superoxide dismutase (SOD), and glutathione S-transferase (GST) activities were observed in the cisplatin-treated rat hepatic tissues. In contrast, PYC treatment effectively prevented cisplatin-induced hepatotoxicity, including the elevation of aminotransferase and histopathological lesions, in a dosedependent manner. Moreover, PYC treatment also induced antioxidant activity by decreasing MDA level and increasing GSH content and SOD and GST activities in liver tissues. These results indicate that PYC has a protective effect against acute hepatotoxicity induced by cisplatin in rats, and that the protective effects of PYC may be due to inhibiting lipid peroxidation and increasing antioxidant activity.

Expression of TGF-beta1/Smad protein in rat liver fibrosis model and the role of IFN-gamma

To study the impact of IFN-gamma on liver fibrosis and its possible mechanism. Thirty healthy male SD rats were randomly divided into two groups: fibrosis model group, IFN-gamma treatment group. Experimental liver fibrosis was induced by subcutaneous injection of CCl4. After 12-week-treatment, serum hyalurnic acid and TGF-beta1 was examined, histopathological changes and degrees of fibrosis were observed by optical microscopy. Meanwhile, the expression of TGF-beta1, TbetaR- I and Smad2/3 proteins was detected by immunohistochemistry and quantified by using computerized image analysis. (1) Pathological observation of hepatic specimens: histological examination showed that there were significant difference between normal group and fibrosis model group by comparing with the degrees of inflammation and fibrosis (P < 0.05). And the difference between fibrosis model group and IFN-gamma treatment group was significant (P < 0.05). (2) Changes of the hepatic fibrosis index (serum HA and TGF-beta1): the levels of serum HA, TGF-beta1 in fibrosis model group were higher than IFN-gamma treatment groups (P < 0.05). (3) Changes of gene protein levels about TGF-beta1/Smad: the expressions of TGF-beta1, TbetaR- I and Smad2/3 in rat hepatic tissue were detected with immunohistochemistry techniques. The expressions of the three items in model group were higher than normal group (P < 0.01). The difference between model group and IFN-gamma treatment group was significant (P < 0.05); IFN-gamma treatment group had significant results on treating experimental hepatic fibrosis. By the way of inhibiting expressions of TGF-beta1, TbetaR- I, Smad2/3, IFN-gamma treatment group exerted its anti-fibrosis effect.

Leucas zeylanica (L.) R. Br. protects ethanol and hydrogen peroxide-induced oxidative stress on hepatic tissue of rats

The aim of the study was to evaluate the effect of Leucas zeylanica against oxidative stress on hepatic tissue. Oxidative stress was induced by exposing hepatic tissue to ethanol and Fenton’s reagent (H2O2+FeSO4). The effect of oxidative stress on liver also was evaluated by the determination of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) activity and the levels of lipid peroxide (LPO). The antioxidative activity of L. zeylanica was determined by estimating it ability to inhibit the hepatic levels of lipid peroxide (LPO), as indicator of oxidative stress. Concomitantly, the antioxidant phytochemicals such as polyphenols and flavonoids were assessed against pyrogallol and quercetin standards. The ALT and AST activities and the levels of LPO of hepatic tissue were significantly increased by oxidative stress. L. zeylanica pretreatment, however, significantly repressed the oxidative stress on hepatic tissue, as indicated by the decreased activities of ALT and AST enzymes and levels of LPO. Analyses of the phytochemicals revealed that the extract of L. zeylanica contained substantial amounts of polyphenols (74.32 ± 4.6 µg of pyrogallol equivalent/mg) and flavonoids (15.69 ± 2.2 µg quercetin equivalent/mg of extract). Finally, the results of the present study demonstrated the presence of antioxidant phytochemicals, including polyphenols and flavonoids in L. zeylanica and henceforth conferred protection against ethanol and H2O2-induced oxidative stress on hepatic tissue.DOI: http://dx.doi.org/10.3329/icpj.v2i9.16076 International Current Pharmaceutical Journal, August 2013, 2(9): 148-151

Expression change of TNF–α in myocardium and hepatic tissue of rats with compound stress of hyperthermia and lipopolysaccharide

ObjectiveTo investigate the expression characteristics of TNF-α in myocardium and hepatic tissue of rats with compound stress of hyperthermia and lipopolysaccharide (LPS). MethodsMale SPF Wistar rats were randomly divided into room temperature+physiological saline group (Group C), hyperthermia+physiological saline group (Group H), room temperature+LPS group (Group L) and hyperthermia+LPS group (Group HL). The rats were put in simulated climate cabin. Group HL and Group H were exposed in the environment at a dry bulb temperature (TDB) of (35.0±0.5) °C, while Group L and Group C were exposed in the environment at a TDB of (26.0±0.5) °C. The rats in Group HL and Group L were given tail intravenous injection of LPS 10 mg/kg, while the rats in Group H and Group C were given tail intravenous injection of 9 g/L NaCl 10 mL/kg. After the stress, immunohistochemical SABC staining method was used to detect the expression characteristics of TNF-α in myocardium and hepatic tissue of rats, and those rats were given routine pathological examinations. ResultsThe expression of TNF-α in myocardium and hepatic tissue in Group HL was enhanced remarkably, and the tissue damages of Group HL were severest. ConclusionsThe eardiotoxieity and hepatotoxicity caused by compound stress of hyperthermia and LPS is closely related to the expression of TNF-α.

English

The present study sought to investigate the effect of ripening on the antioxidant properties and phytochemical constituents of plantain, a major staple food of global consumption. This was done by measuring the total phenolics and flavonoid content of both ripe and unripe form of plantain. Moreover, the free radical scavenging, ferric reducing and iron chelating properties of both forms of plantain were also investigated. Finally, the ability of both extracts to prevent pro-oxidant induced deoxyribose degradation and lipid peroxidation in both cerebral and hepatic tissues of rat was also assessed. Results revealed that ripening depleted the total phenolics [(unripe, 16.2 &plusmn; 0.75 mg/g (GAE); ripe, 8.3 &plusmn; 0.55 mg/g (GAE)] and flavonoid [(unripe, 15.2 &plusmn; 0.45 mg/g (QE); ripe, 9.5 &plusmn; 0.21 mg/g (QE) content of plantain. Moreover, the unripe plantain extract exhibited a significantly higher free radical scavenging, iron (II) chelating and ferric reducing effect than the ripe extract. Finally, the unripe plantain extract offered a marked inhibitory effect against pro-oxidant induced deoxyribose degradation and lipid peroxidation than the ripe extract. Put together, we could suggest that the observed depletion in antioxidant properties of ripe plantain may be due to an irreversible degradation in the phytochemical constituents of the fruit as ripening progresses. This study would create a public awareness on the effect of ripening on the therapeutic potentials of plantain and serve as useful information in the global fight against degenerative diseases whose etiology has been linked to oxidative stress. &nbsp; Key words:&nbsp;Musa paradisiaca, free radical, oxidative stress, phytochemical, ripening.

Bioactivation of nevirapine to a reactive quinone methide: implications for liver injury.

Nevirapine (NVP) treatment is associated with a significant incidence of liver injury. We developed an anti-NVP antiserum to determine the presence and pattern of covalent binding of NVP to mouse, rat, and human hepatic tissues. Covalent binding to hepatic microsomes from male C57BL/6 mice and male Brown Norway rats was detected on Western blots; the major protein had a mass of ∼55 kDa. Incubation of NVP with rat CYP3A1 and 2C11 or human CYP3A4 also led to covalent binding. Treatment of female Brown Norway rats or C57BL/6 mice with NVP led to extensive covalent binding to a wide range of proteins. Co-treatment with 1-aminobenzotriazole dramatically changed the pattern of binding. The covalent binding of 12-hydroxy-NVP, the pathway that leads to a skin rash, was much less than that of NVP, both in vitro and in vivo. An analogue of NVP in which the methyl hydrogens were replaced by deuterium also produced less covalent binding than NVP. These data provide strong evidence that covalent binding of NVP in the liver is due to a quinone methide formed by oxidation of the methyl group. Attempts were made to develop an animal model of NVP-induced liver injury in mice. There was a small increase in ALT in some NVP-treated male C57BL/6 mice at 3 weeks that resolved despite continued treatment. Male Cbl-b–/– mice dosed with NVP had an increase in ALT of >200 U/L, which also resolved despite continued treatment. Liver histology in these animals showed focal areas of complete necrosis, while most of the liver appeared normal. This is a different pattern from the histology of NVP-induced liver injury in humans. This is the first study to report hepatic covalent binding of NVP and also liver injury in mice. It is likely that the quinone methide metabolite is responsible for NVP-induced liver injury.

Food restriction and refeeding induces changes in lipid pathways and fat deposition in the adipose and hepatic tissues in rats with diet‐induced obesity

The aim of this study was to determine the effects of successive cycles of a moderately restrictive diet and refeeding with a high-fat diet on the metabolism of the adipose and hepatic tissues of obese rats. Rats were assigned to the following groups: a chow diet; a high-fat diet; a moderate caloric restriction; or a moderate caloric restriction plus refeeding. Some animals in each group were given [1-(14)C]triolein intragastrically, while others received an intraperitoneal injection of 3 mCi (3)H(2)O. All animals were killed by decapitation. The retroperitoneal, visceral epididymal and omental white adipose tissues, brown adipose tissue, liver and blood were immediately removed. The lipid uptake from the diet, in vivo rate of lipogenesis, percentage of fat, lipid profile and leptin concentration were analysed. The high-fat diet promoted an increase in fatty liver (P ≤ 0.05), adiposity mass (P ≤ 0.05) and the plasma concentration of leptin (P ≤ 0.05) and a decreased lipid uptake in white adipose tissue depots (P ≤ 0.05) in relation to the chow diet. The moderate caloric restriction did not reverse the changes promoted by the high-fat diet but induced a small decrease in adiposity, which was reversed after refeeding, and the animals maintained a dyslipidaemic profile and high fat deposition in the liver. We can conclude that the high-fat diet and subsequent moderate caloric restriction plus refeeding increased the risks of developing visceral obesity, dyslipidaemia and non-alcoholic fatty liver disease, which suggests that this type of experimental protocol can be used to study mechanisms related to the metabolic syndrome.

Effect of boiling on the phytochemical constituents and antioxidant properties of African pear Dacryodes edulis seeds in vitro

African pear, Dacryodes edulis, seed extract has been used for the treatment of ailments in traditional medicine. However, the effect of boiling on its antioxidant properties is still poorly understood. Therefore, the present study investigates the effect of boiling on the antioxidant properties of D. edulis seed extract using in vitro parameters such as free radical scavenging ability against 2, 2- diphenyl -1- picrylhydrazyl (DPPH) radical, iron (III) reducing and iron (II) chelating ability. Furthermore, the ability of both extracts (boiled and unboiled) to offer protective benefit against lipid peroxidation in cerebral and hepatic tissues of rat was assessed. Moreover, the effect of boiling on the phytochemical constituents (total phenolics, flavonoids and vitamin C) of the seed extracts was measured. Results indicate that boiling significantly (P < 0.05) potentiates the total phenolic [(Boiled 60.1 ± 0.88 mg/g (GAE); Unboiled 30.2 ± 0.68 mg/g (GAE)] and flavonoid [(Boiled: 50.02 ± 0.12 mg/g (QE); Unboiled: 35.8 ± 0.15 mg/g (QE)] content but mildly depleted the vitamin C content [Boiled: (36.9 ± 0.44 mg/g; Unboiled: 40.1 ± 0.21 mg/g]. Similarly, boiling markedly increased the antioxidant properties (free radical scavenging, iron (II) chelating, iron (III) reducing and inhibitory effect against pro-oxidant-induced lipid peroxidation) of the seed extract. From the foregoing, the wide usage of African pear as remedy for ailment in folk medicine may be due to its phytochemical constituents which are potentiated by boiling. Hence, information from this study would create public awareness especially to traditional medical practitioners who are involved in the act of boiling the fruit to get the extract used for medicinal purposes. Key words: African pear, phytochemical, degradation, proxidant, antioxidant, mechanism, lipid peroxidation.

Increase in covalent binding of 5-hydroxydiclofenac to hepatic tissues in rats co-treated with lipopolysaccharide and diclofenac: involvement in the onset of diclofenac-induced idiosyncratic hepatotoxicity

Diclofenac (DCF), a nonsteroidal anti-inflammatory drug, is well known to induce idiosyncratic hepatotoxicity. Although there remains much to be elucidated about its onset mechanism, it is widely accepted as a hypothesis that idiosyncratic hepatotoxicity arises from a specific immune response to a hapten formed by covalent binding of drugs or their reactive metabolites to hepatic tissues. In this study, we investigated the effects of covalent binding of DCF reactive metabolites to hepatic tissues using a rat model of liver injury induced by co-treatment with lipopolysaccharide (LPS) at a non-hepatotoxic dose. In studies done in vitro using hepatic microsomes prepared from rats treated with LPS alone, 4'- and 5-hydroxylation activities on DCF metabolism and adducts of reactive metabolites to dansyl glutathione (dGSH) were markedly decreased associated with a decrease in total P450 content. However, in studies done in vivo, the LPS/DCF co-treatment significantly increased adducts of 5-hydroxydiclofenac (5-OH-DCF) to rat hepatic tissues and delayed the elimination of 5-OH-DCF from plasma. Furthermore, we investigated the effects of co-treatment on hepatic GSH level in rats. A decrease of hepatic GSH was observed with the LPS/DCF co-treatment but not with LPS or DCF alone. The results suggest that covalent binding of reactive metabolites via 5-OH-DCF to hepatic tissues may play an important role in the onset of DCF-induced idiosyncratic hepatotoxicity, especially under decreased GSH conditions.

Fractionation of acacia honey affects its antioxidant potential in vitro

ObjectiveTo investigate the effects of fractionation of acacia honey on its antioxidant potential in contrast with the pure honey from whole blood, brain and liver in vitro. MethodsHoney was partitioned into three fractions (dichloromethane, ethyl acetate and aqueous). Their immuno-modulatory effect on whole blood was assayed using Luminol-amplified chemiluminescence technique. Their antioxidant activities on rat brain and hepatic tissues which covers for catalase, SOD activities and lipid peroxidation. ResultsFractions of the honey enhanced the production of radicals with no significant (P>0.05) antioxidant activity on whole blood where as pure honey does. Pure honey significantly (P<0.05) stimulates SOD and Catalase activity with no significant (P>0.05) effects on lipid peroxidation. ConclusionsFractionation of acacia honey negatively affects its antioxidant potential thereby making it a radical generating agent in contrast with the unfractionated.

Serotonin modulates hepatic 6-phosphofructo-1-kinase in an insulin synergistic manner

Human and rat hepatic tissue express many serotonin (5-HT) receptor subtypes, such as 5-HT1B, 5-HT2A, 5-HT2B and 5-HT7 receptors, which mediate diverse effects. 5-HT is known to regulate several key aspects of liver biology including hepatic blood flow, innervations and wound healing. 5-HT is also known to enhance net glucose uptake during glucose infusion in fasted dogs, but little is known about the ability of 5-HT to control hepatic glucose metabolism, especially glycolysis. This study addresses the potential of 5-HT to regulate PFK activity and the mechanisms related to the enzyme activity. Based on our results, we are the first to provide evidence that 5-HT up-regulates PFK in mouse hepatic tissue. Activation of the enzyme occurs through the 5-HT2A receptor and phospholipase C (PLC), resulting in PFK intracellular redistribution and favoring PFK association to the cytoskeletal f-actin-enriched fractions. Interestingly, 5-HT and insulin act in a synergistic manner, likely because of the ability of insulin to increase fructose-2,6-bisphosphate because the presence of this PFK allosteric regulator enhances the 5-HT effect on the enzyme activity. Together, these data demonstrate the ability of 5-HT to control hepatic glycolysis and present clues about the mechanisms involved in these processes, which may be important in understanding the action of 5-HT during the hepatic wound healing process.

Increase of the Activity of Phase II Antioxidant Enzymes in Rats after a Single Dose of Coffee

This study evaluated the acute effect of the administration of coffee brew in the activity of phase II antioxidant enzymes in the hepatic tissue of rats. A single dose of this beverage increased the activity of the enzymes SOD, CAT, and GPx; the maximum increase occurred 1 h after administration (19.1, 22.1, and 25.1%, respectively). These changes were statistically significant (p < 0.05), the response was shown to be dose-dependent (p < 0.05), and the return to basal levels took >4 h from the intervention, suggesting a long-term effect. The total antioxidant capacity of the hepatic tissue also exhibited a peak 1 h after the intervention (6.5%), but the increase was not statistically significant and the response was not dose-dependent due to the low exposure to coffee. These results indicate that coffee increases the activities of antioxidant enzymes, improving protection against oxidative stress.

Effects of Chinese herbal medicine Yiguanjian Decoction on collagen metabolism of hepatic tissues in rats with CCl4-induced liver fibrosis

To investigate the effects of Yiguanjian Decoction, a compound traditional Chinese herbal medicine, on collagen metabolism of hepatic tissues in rats with carbon tetrachloride (CCl(4))-induced liver fibrosis. Liver fibrosis was induced in rats by intraperitoneal injection of 50% CCl(4)-olive oil solution at a dose of 1 mL/kg body weight, twice per week for 9 consecutive weeks. Six rats were sacrificed for dynamic observation at the end of the 3rd and 6th week respectively, and the other rats were divided into 9-week untreated group and Yiguanjian Decoction group which was given Yiguanjian Decoction intragastrically in the subsequent 3-week modeling period. Another 6 rats were used as normal group. Rats in the normal group and 9-week untreated group were treated with distilled water. At the end of the 9th week, all rats were sacrificed, and their blood serum and liver tissue were collected for measuring hepatic histology and expressions of α-smooth muscle actin (α-SMA), tissue inhibitor of metalloproteinase (TIMP)-1, TIMP-2, matrix metalloproteinase (MMP)-13, MMP-14, collagen type I (Col I), and activities of MMP-2 and -9. Compared with the normal group, collagen fiber deposition, expressions of α-SMA, Col I, TIMP-1, TIMP-2, MMP-13 and MMP-14 and activities of MMP-2 and -9 in the liver tissues gradually increased in the untreated group (P<0.05, P<0.01). These changes were significantly suppressed by Yiguanjian Decoction. Yiguanjian Decoction exerts inhibition on formation of CCl(4)-induced cirrhosis in rats. The therapeutic mechanism may be related to inhibiting hepatic stellate cell activation, collagen secretion, and promoting collagen fiber degradation.

Wheatgrass and Antioxidant Levels in Carbon Tetrachloride-induced Hepatotoxicity in Rats

© 2011, INASL 43 Wheatgrass and Antioxidant Levels in Carbon Tetrachloride-induced Hepatotoxicity in Rats JK Kamboj*, SV Rana*, RP Ola*, DK Dhawan**, K Vahiphei*** Departments of Gastroenterology and ***Pathology, PGIMER, Chandigarh Background: Wheatgrass (Triticum aestivum), rich in various antioxidants, is known to treat a number of conditions including burns, common cold, anemia, cough, fever and eczema, etc. Aim: Therefore, this study was planned to determine the effect of wheatgrass on antioxidant levels and oxidative stress induced by CCl4 in hepatic tissue of rats. Materials and Methods: Eighteen Wistar rats with body weight of 120–150 gms were divided into 3 groups. Group 1—rats in this group were given normal saline injection subcutaneously twice a week for 4 weeks. Group 2—carbon tetrachloride was administered subcutaneously at a dose of 2 mL/Kg body weight twice a week for 4 weeks in this group. Group 3—wheatgrass was administered orally at 80 mg/100 g body weight in drinking water for 4 weeks along with CCl4 injection given subcutaneously twice a week at a dose of 2 mL/Kg body weight wheatgrass dose was started 2 weeks prior to first injection of CCl4. Reduced glutathione (GSH) was measured in liver tissue for antioxidant levels and lipid peroxidation (LPO) for oxidative stress. Results: GSH level was significantly decreased while LPO increased in hepatic tissue of group 2 (CCl4 treated rats) as compared to group 1 (controls). In group 3 (CCl4 + wheatgrass treated rats), GSH level was increased while LPO decreased as compare to group 2. Conclusion: This study shows that wheatgrass treatment decreased the oxidative stress and increased the antioxidant levels in CCl4 induced hepatotoxicity. Conflict of Interest: None Comparative Hepatoprotective Activity of Polygonum Bistorta Extract and Its Active Compound Against Carbon Tetrachloride-Induced Hepatotoxicity in Rats DK Mittal School of Studies in Zoology, Jiwaji University, Gwalior, India Aim: Many herbal preparations have been recommended in alternative system of medicine for the treatment of hepatic disorder. Druginduced hepatotoxicity is still a significant unresolved clinical problem as liver is the most common site of damage. Present findings is based on the protective effect of PB extract and TA against acute and sub chronic APAP and CCl4 intoxication. Methods: Male Sprague Dawley strain rats (150 ± 10 g body weight) were administered a single bolus dose of APAP (2 g/Kg, p.o.) and CCl4 (0.15 mL/Kg, i.p.). Plant extract (Polygonum bistorta) and its active principle (tannic acid) was given at the dose of 100 mg/Kg and 25 mg/Kg, respectively (p.o.) after 24 hours of toxicant administration. Results: The effect of PB, TA and silymarin on AST, ALT, SALP, Blood sugar, Protein, Albumin, Bilirubin, Urea, Creatinine were observed against APAP toxicity. Further, the effects of the PB and TA was confirmed on LPO, GSH, ATPase, SOD, GR, GPx, G6PDH, GST, AH, AND were estimated against CCl4 intoxication. Conclusions: TA produced significant hepatoprotective effect by decreasing the activities of above biochemical indices followed by PB extract. Histopathological and ultrastructural observations and in vitro study by MTT assay supported above findings. Conflict of Interest: None 03_JCEH-Abstract.indd 43 3/18/2011 11:13:05 AM

Comparative Hepatoprotective Activity of Polygonum Bistorta Extract and Its Active Compound Against Carbon Tetrachloride-Induced Hepatotoxicity in Rats

© 2011, INASL 43 Wheatgrass and Antioxidant Levels in Carbon Tetrachloride-induced Hepatotoxicity in Rats JK Kamboj*, SV Rana*, RP Ola*, DK Dhawan**, K Vahiphei*** Departments of Gastroenterology and ***Pathology, PGIMER, Chandigarh Background: Wheatgrass (Triticum aestivum), rich in various antioxidants, is known to treat a number of conditions including burns, common cold, anemia, cough, fever and eczema, etc. Aim: Therefore, this study was planned to determine the effect of wheatgrass on antioxidant levels and oxidative stress induced by CCl4 in hepatic tissue of rats. Materials and Methods: Eighteen Wistar rats with body weight of 120–150 gms were divided into 3 groups. Group 1—rats in this group were given normal saline injection subcutaneously twice a week for 4 weeks. Group 2—carbon tetrachloride was administered subcutaneously at a dose of 2 mL/Kg body weight twice a week for 4 weeks in this group. Group 3—wheatgrass was administered orally at 80 mg/100 g body weight in drinking water for 4 weeks along with CCl4 injection given subcutaneously twice a week at a dose of 2 mL/Kg body weight wheatgrass dose was started 2 weeks prior to first injection of CCl4. Reduced glutathione (GSH) was measured in liver tissue for antioxidant levels and lipid peroxidation (LPO) for oxidative stress. Results: GSH level was significantly decreased while LPO increased in hepatic tissue of group 2 (CCl4 treated rats) as compared to group 1 (controls). In group 3 (CCl4 + wheatgrass treated rats), GSH level was increased while LPO decreased as compare to group 2. Conclusion: This study shows that wheatgrass treatment decreased the oxidative stress and increased the antioxidant levels in CCl4 induced hepatotoxicity. Conflict of Interest: None Comparative Hepatoprotective Activity of Polygonum Bistorta Extract and Its Active Compound Against Carbon Tetrachloride-Induced Hepatotoxicity in Rats DK Mittal School of Studies in Zoology, Jiwaji University, Gwalior, India Aim: Many herbal preparations have been recommended in alternative system of medicine for the treatment of hepatic disorder. Druginduced hepatotoxicity is still a significant unresolved clinical problem as liver is the most common site of damage. Present findings is based on the protective effect of PB extract and TA against acute and sub chronic APAP and CCl4 intoxication. Methods: Male Sprague Dawley strain rats (150 ± 10 g body weight) were administered a single bolus dose of APAP (2 g/Kg, p.o.) and CCl4 (0.15 mL/Kg, i.p.). Plant extract (Polygonum bistorta) and its active principle (tannic acid) was given at the dose of 100 mg/Kg and 25 mg/Kg, respectively (p.o.) after 24 hours of toxicant administration. Results: The effect of PB, TA and silymarin on AST, ALT, SALP, Blood sugar, Protein, Albumin, Bilirubin, Urea, Creatinine were observed against APAP toxicity. Further, the effects of the PB and TA was confirmed on LPO, GSH, ATPase, SOD, GR, GPx, G6PDH, GST, AH, AND were estimated against CCl4 intoxication. Conclusions: TA produced significant hepatoprotective effect by decreasing the activities of above biochemical indices followed by PB extract. Histopathological and ultrastructural observations and in vitro study by MTT assay supported above findings. Conflict of Interest: None 03_JCEH-Abstract.indd 43 3/18/2011 11:13:05 AM