Catalase Activity In Liver Tissues Research Articles

An efficient protocol for quantifying catalase activity in biological samples

BackgroundCatalase is an important enzyme that helps protect cells against oxidative stress. The current protocol presents a reliable method for measuring catalase (CAT) enzyme activity in biological systems using the CUPRAC-CAT method.MethodsIn the CUPRAC-CAT method, the component of the enzymatic reaction was incubated before adding the Cu(Nc)22+ reagent. The unreacted substrates reduced the Cu(II)-the neocuproine complex, resulting in the highly colored Cu(I)-neocuproine product, which could be detected spectrophotometrically at 450 nm. The negative correlation between catalase activity and the absorbance of the Cu(I)-neocuproine complex was examined. To assess the optimization of Cu(I)-neocuproine complex production, response surface methodology (RSM) was employed, specifically utilizing the Box–Behnken design (BBD). Additionally, the reliability of the newly developed protocol was confirmed through Bland–Altman analysis of catalase activity in paired samples, employing the peroxovanadate method.ResultsThe novel method is just as accurate as the established standard; the correlation between the two methods was 0.99. The CUPRAC-CAT method is stable, sensitive, linear, reproducible, accurate, and selective and can be used for quantifying oxidative stress while measuring catalase activity in liver tissue homogenates.ConclusionsThis study has demonstrated a straightforward and dependable protocol for evaluating catalase activity. The protocol is free from interference and can be easily employed in scientific research, ensuring a high level of accuracy and precision. The CUPRAC-CAT method is an effective technique to monitor bacterial contamination. This method provides quick and reliable results that can help ensure food safety and prevent or address bacterial contamination.Graphical abstract

GENDER DIFFERENCES IN THE EFFECT OF ANTIULCER DRUGS AND PLACENTA CRYOEXTRACT ON THE INTENSITY OF LIPID PEROXIDATION AND THE ACTIVITY OF THE ANTIOXIDANT SYSTEM IN EXPERIMENTAL HEPATITIS WITH ETHANOL-INDUCED CIRRHOSIS

Introduction. It is well recognized that drug metabolism products in the liver can induce oxidative stress and mitochondrial dysfunction, leading to the development of hepatocellular injury. As a potential agent capable of counteracting the hepatotoxic effects of drugs, we focused our attention on a domestic biotechnological preparation – cryopreserved placental extract (CPE).
 The aim. To characterize gender differences in the effect of esomeprazole, clarithromycin, metronidazole (E/C/M), and CPE on the intensity of lipid peroxidation and the activity of the antioxidant system in tetrachloromethane (CCl4) hepatitis with a background of ethanol-induced cirrhosis (ETCM).
 Materials and methods. The study was conducted with varying levels of sex hormones on 112 male and female rats. Chronic ETCM was induced by administering a 50.0% oil solution of CCl4 at a dose of 8 ml/kg body weight of the animals twice a week, in combination with a 5.0% ethanol solution for drinking over a period of 45 days. The content of TBA-RP in liver homogenates was determined spectrophotometrically by the method described by Asakawa T. et al. Catalase activity in liver homogenates was determined spectrophotometrically according to the method of Korolyuk M.A. and co-authors.
 Results. The most pronounced increase in lipid peroxidation processes was observed in females with chronic ETCM-induced liver damage and administration of antiulcer drugs following ovariectomy, resulting in a TBA-RS content of 36.1±2.79 μmol/kg of tissue. Administration of E/C/M in animals with chronic liver damage led to a suppression of the antioxidant system, as evidenced by a decrease in catalase activity in liver tissues.
 Conclusion. The combined use of anti-ulcer drugs and CPE on the background of chronic ETCM mitigated the activation of lipid peroxidation processes, which was indicated by a statistically significant (p < 0.001) 2.7-fold lower content of TBA-RP in liver homogenates. Additionally, it was established that the administration of CPE was accompanied by a statistically significant increase in catalase activity in females, more prominently than in males. In females without changes in hormonal status, the introduction of CPE resulted in a growth (p < 0.001) of catalase activity by 75.0%, with the most significant increase observed in females after ovariectomy – catalase activity statistically significantly (p < 0.001) increased by 2.6 times compared to the indicators of females not administered with CPE. The administration of CPE in female rats without altering hormonal status was accompanied by a twofold (p < 0.01) increase in the antioxidant-prooxidant index compared to male rats, indicating more pronounced antioxidant properties of CPE in female rats.

Alterations in blood parameters, DNA damage, oxidative stress and antioxidant enzymes and immune-related genes expression in Nile tilapia (Oreochromis niloticus) exposed to glyphosate-based herbicide

In this study, effects of different concentrations (0, 5, 10, 20, 30, and 40 mg/L) of glyphosate-based herbicide (GBH) on Nile tilapia (Oreochromis niloticus) were investigated after a 14-day of exposure. After determination of LC50 value, effects of GBH on hematological and serum biochemical parameters in blood samples, DNA damage, lipid peroxidation and catalase activity in liver tissues, expression levels of antioxidant enzyme-related genes (SOD, CAT, GPx, and GST) and immune-related genes (TGF-β, TGF-α and IL1-β) were evaluated. The LC50 value has been found as 44.4 mg/L for GBH. GBH exposure at all concentrations caused alterations in blood parameters. GBH induced oxidative stress in liver and DNA damage in blood. Antioxidant enzyme-related genes were significantly up-regulated to suppress oxidative stress. On the other hand, the expression levels of immune-related genes decreased in fish exposure to particularly ≥20 mg/L GBH.

Effects of Centella asiatica extract on antioxidant status and liver metabolome of rotenone-treated rats using GC-MS.

Centella asiatica has been used as a culinary vegetable or medicinal herb. In this study, the hepatoprotective effect of the standardized extract of C. asiatica (ECa233) in rotenone-treated rats was examined using a GC-MS-based metabolomic approach. ECa233 contains >80% triterpenoids with a ratio of madecassoside to asiaticoside of 1.5(±0.5):1. Rats were randomly divided into three groups (with six rats/group): sham negative control, rotenone positive control and the ECa233 test group. Rats in the ECa233 group received 10 mg/kg ECa233 orally for 20 days, followed by 2.5 mg/kg intraperitoneal rotenone injection to induce toxicity before being sacrificed. Metabolomic analysis showed that supplementation of ECa233 protected rat liver against rotenone toxicity. Pipecolinic acid was one of the most important metabolites; its level was decreased in the rotenone group as compared with the control. Supplementation with ECa233 before administration of rotenone raised pipecolinic acid to levels intermediate between controls and rotenone alone. The metabolomics approach also helped discover a possible new genuine epimetabolite in the present work. Antioxidant tests revealed that ECa233 inhibited lipid peroxidation and increased catalase activities in liver tissue.

Propolis alleviates concanavalin A-induced hepatitis by modulating cytokine secretion and inhibition of reactive oxygen species.

Viral hepatitis-induced oxidative stress accompanied by increased levels of transforming growth factor-β (TGF-β) and hepatic fibrosis are hallmarks of hepatitis C virus infection. The present study was designed to investigate the potential protective effect of propolis against liver injury induced by concanavalin A (Con A), a T-cell-dependent model that causes an immune-mediated hepatitis in a similar pattern to the one induced by viral infections. In the present study, rats were randomly divided into four groups. The first group (control) was administered the vehicle of Con A (i.v.) for 24h. The second group received Con A (12mg/kg body weight i.v.) for 24h. The third group received propolis (300mg/kg by oral gavage) 5days before and concurrently with Con A for 24h. The last group received propolis alone. Following a single injection of Con A, histopathological changes as well as significant reduction in albumin level were observed. In addition, serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and total bilirubin were significantly increased. These increases correlated with an increase in lipid peroxidation and downregulation of reduced glutathione (GSH) as well as superoxide dismutase (SOD) and catalase activities in liver tissue. Furthermore, these changes were associated with an increase in serum levels of the inflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) as well as the profibrotic cytokine TGF-β. Moreover, TGF- β activation was accompanied with an increase in Smad phosphorylation. Interestingly, concomitant administration of propolis along with Con A significantly attenuated all these negative effects and improved liver function indicating that propolis has the ability to protect rats from Con A-induced hepatitis.

Thaumatococcus daniellii Benn. (Marantaceae) Leaf Methanol Extract Possessed Hepatoprotective Effect Against Acetaminophen-induced Liver Injury in Albino Rats

[Background]Acetaminophen or Paracetamol is widely used as analgesic and antipyretic or antimalarial drug, but at high dose it leads to undesirable side effects, such as hepatotoxicity. [Objective]This present study demonstrates the in vivo hepatoprotective effect of Thaumatococcus daniellii against acetaminophen induced liver toxicity. [Methods] Acetaminophen- induced liver injury was evaluated by an increase in serum AST (or SGOT), ALT (or SGPT), ALP activity and bilirubin level accompanied by significant decrease in albumin level. [Results] Acetaminophen hepatoxicity was manifested by an increase in lipid peroxidation, depletion of reduced glutathione (GSH) and catalase activity in liver tissue. Oral administration of the plant extract protects the rats against acetaminophen induced liver injury by increased lipid peroxidation, restored altered serum marker enzymes and antioxidant level to normal liver morphology.[Conclusion]The results showed that T. daniellii leaf extract displayed significant liver healing efficacy against acetaminophen induced hepatotoxicity in rats. The result further affirm the use of the plant as an ethno-medicinal prescription for liver related injury.

Biochemical Studies on Evaluation of New Composite against Hepatocellular carcinoma-induced in Rats: Recent Therapeutic Approaches

The hepatoprotective effect of some natural and synthetic compounds against chemically inducedhepatocellular carcinoma in rats was evaluated. One hundred male albino rats were divided equally intofive groups. Normal control group, carcinogenic [Ferric nitrilotriacetate (Fe-NTA: 9 mg Fe/kg b.wt. i.p.)and chloroform (150 mg/ kg b.wt. orally)] - induced group, curcumin group (400 mg/kg. b.wt. orally),tetrachlorocuprate-lysine (25 mg/kg. b. wt. s.c.) and ascorbate (500 mg/kg. b. wt. orally) group and amixture group (composed of curcumin, tetrachlorocuprate -lysine and ascorbate). Blood samples andliver tissue specimens were collected at the end of experiment (4 months) for determination of thefollowing parameters: Tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1), catalase andmyeloperoxidase (MPO) in liver tissues, in addition to serum malondialdehyde (MDA) and reducedglutathione (GSH). Moreover, histopathological examination of liver tissues was done for resultsconfirmation. The obtained results showed a significant elevation in MDA, MPO and immunologicalmarkers levels, with significant reduction in serum reduced glutathione and catalase activity in livertissue in hepatocellular-carcinogen induced rats as compared to the control group. However,administration of rats with the compounds under investigation resulted in a significant reduction ofMDA, MPO and immunological markers levels, and increased in reduced glutathione and catalase levelscompared to the carcinogenic non treated group. Various pathological alterations were observed in liverof chemically induced-carcinogenic group interestingly, results supported the protective effect of thecompounds under investigation and preserved the histological structures of liver tissues. These resultsconcluded that basic curcumin, tetrachlorocuprate-lysine and ascorbate exert chemopreventative effectagainst hepatocellular carcinoma.

Hepatoprotective effect of Livplus-A polyherbal formulation

Objective: The aim of the present study was to investigate the hepatoprotective effect of Livplus (a polyherbal formulation) against CCl4-induced hepatotoxicity in rats. Methods: Hepatotoxicity was induced in rats by i.p. injection of CCl4 once three days for 14 days. Livplus or Silymarin was administered along with CCl4 and the biochemical parameters like aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkalinephosphatase (ALP), total bilirubin (TB), direct bilirubin, total protein (TP), gamma-glutamyl transferase (GGT), total cholesterol (TC) and triglycerides (TG) were estimated. Furthermore, biomarkers of oxidative stress such as MDA levels, Glutathione contents, SOD and catalase activity in liver tissue were estimated. Results: Treatment with Livplus significantly reduced the elevated levels of ALT, AST, ALP, bilirubin (direct and total), GGT, TC, TG and increased levels of TP compared to CCl4 control rats. The treatment with Livplus also showed a significant increase in glutathione contents, SOD and catalase activity and a decrease in MDA levels compared to CCl4 control rats. Conclusion: The finding of present study indicates that Livplus showed a potential hepatoprotective activity. These results support the traditional use of Livplus in the treatment of liver disorders.

Effects of dietary zinc oxide nanoparticles on growth performance and antioxidative status in broilers.

Broilers in four groups were fed a basal diet supplemented with 60 mg/kg zinc oxide (60-ZnO; control), or 20, 60, or 100 mg/kg ZnO nanoparticles (20-, 60-, and 100-nano-ZnO, respectively). Compared with the controls, after 14 days, birds in the 20- and 60-nano-ZnO groups had significantly greater weight gains and better feed conversion ratios. However, the body weight of birds in the 100-nano-ZnO group was dramatically reduced after 28 days. Relative to the control group, the total antioxidant capability (T-AOC) in serum and liver tissue was significantly higher in the 20-nano-ZnO group at all time points and also significantly higher in the 60- and 100-nano-ZnO groups in serum on days 28 and 35 and in liver tissues on days 21 and 28. Compared with the controls, the activity of copper-zinc superoxide dismutase (Cu-Zn-SOD) was significantly greater in the 60- and 100-nano-ZnO groups in serum on days 28 and 35 and in liver tissues after 21 days. Catalase activity in serum samples was significantly higher in the 20- and 60-nano-ZnO groups relative to the control and 100-nano-ZnO birds, but catalase activity in liver tissue was not affected by different nano-ZnO levels. Malondialdehyde content in serum and liver tissues was significantly reduced in the 20-, 60-, and 100-nano-ZnO groups compared with that in the control group at all time points except day 42. Taken together, our data indicate that appropriate concentration of dietary ZnO nanoparticles improves growth performance and antioxidative capabilities in broilers, and 20 mg/kg nano-ZnO is the optimal concentration.

Evaluation of hepatoprotective activity of abelmoschus moschatus seed in paracetamol induced hepatotoxicity on rat

Paracetamol (PCM) is widely used as analgesic and antipyretic drug, but at high dose it leads to undesirable side effects, such as hepatotoxicity. This study gives the information about hepatoprotective activity of abelmoschus moschatus seed extract against paracetamol and ethanol induced hepatotoxicity. Paracetamol induce hepatotoxicity was evaluated by an increase (P<0.05) in serum AST, ALT, ALP activity and bilirubin level Paracetamol hepatotoxicity was manifested by an increase (P<0.05) lipid peroxidation, depletion of reduced glutathione (GSH) and catalase activity in liver tissue. Administration of etanolic as well as aqueous plants extract [300mg/kg body weight of rat] protects the paracetamol induced lipid peroxidation, restored altered serum marker enzymes and antioxidant level towards normal. The results indicate hepatoprotective activity of all studied plants extract against paracetamol induced toxicity. Ethanolic extract was found more significant than the aqueous extract.

Antihypertensive and antioxidant effects of the Graptopetalum paraguayense E. Walther extract in spontaneously hypertensive rats

AbstractBACKGROUND: Our previous in vitro study showed that 50% ethanolic extract of Graptopetalum paraguayense E. Walther (GE50) exhibited an inhibitory effect on angiotensin I converting enzyme (ACE). This study was aimed at evaluating the antihypertensive and antioxidant effects of GE50 extract on spontaneously hypertensive rats (SHR).RESULTS: Daily oral administration of GE50 extract (2.5 g kg−1 body weight) for 4 weeks exhibited a significant decrease in blood pressure as well as ACE activity of tested tissues in SHR rats, while no significant change was found in normotensive Wistar Kyoto rats (WKY). For SHR, GE50 extract increased the total antioxidant status in the plasma and decreased malondialdehyde levels in all tested tissues. Furthermore, GE50 extract increased α‐tocopherol and glutathione levels in brain tissue, glutathione peroxidase and catalase activities in heart, as well as catalase activity in liver tissue. In WKY rats, GE50 extract only increased ascorbic acid level in liver tissue.CONCLUSION: The present study demonstrated that GE50 might serve as an antioxidant and an ACE inhibitor in convalescence from hypertension in rats. Copyright © 2009 Society of Chemical Industry

Effects of Antioxidant Vitamins on Glutathione Depletion and Lipid Peroxidation Induced by Restraint Stress in the Rat Liver

Stress as a cofactor has been reported to affect the progression and severity of several diseases. The influence of stress on the liver is of interest from the clinical point of view because stress plays a potential role in aggravating liver diseases in general and hepatic inflammation in particular, probably through generation of reactive oxygen species. The present study was undertaken to investigate the potential of the antioxidant vitamins A (retinol), E (tocopherol) and C (ascorbic acid) individually and in combination (vitamin E + C) to modulate restraint stress-induced oxidative changes. These effects were determined by measuring changes in hepatic levels of free radical scavenging enzymes such as superoxide dismutase (SOD), glutathione-S-transferase (GST) and catalase, as well as levels of total glutathione (GSH), malondialdehyde (MDA), aspartate aminotransferase (AST) and alanine aminotransferase (ALT). Immobilisation was achieved by placing the animals in wire mesh cages of their size. The rats were orally administered vitamins A, E and C individually and in combination (E + C) prior to and after 6 hours of immobilisation stress exposure. The hepatic levels of SOD, GST, catalase, GSH and MDA were determined by spectrophotometric methods. Liver SOD activity was assayed by monitoring the amount of enzyme required to inhibit autoxidation of pyrogallol by 50%. Hepatic GST was monitored by following the increase in absorbance at 340 nm of CDNB-GSH conjugate generated due to GST catalysis between GSH and CDNB. Catalase activity in liver tissues was determined using peroxidase as the substrate. Lipid peroxidation was measured by determining the level of thiobarbituric acid reactive substances. ALT and AST were determined by commercial kits. Six hours of immobilisation stress caused a decrease in liver levels of SOD (p = 0.001), catalase (p = 0.031), GST (p = 0.021) and GSH (0.013), while levels of MDA (p = 0.0015), AST (p = 0.05) and ALT (p = 0.046) were increased compared with non-stressed control rats. Both pre-vitamin stress and post-vitamin stress treatments either alone or in combination were associated with increased normalisation of these parameters towards control values, with post-vitamin treatment being the more effective of the two. Vitamins E and C individually were found to be more effective in restoring the endogenous antioxidant system than vitamin A. The combined vitamin (E + C) post-stress treatment was found to be effective but not additive in combating hepatic oxidative stress. The beneficial effects of these vitamin treatments were also reflected in reversions of altered AST and ALT levels towards their control values. Vitamins E or C alone or in combination can be given as prophylactic/therapeutic supplements for combating scavenging free radicals generated in liver tissue. This approach may reduce oxidative stress caused by diseases such as cirrhosis.

Potential effects of L-NAME on alcohol-induced oxidative stress

Nitric oxide (NO) is a highly reactive oxidant synthesized from L-arginine by nitric oxide synthase (NOS). NO may cause injury through the generation of potent radicals. Nw- nitro-L-arginine methyl ester (L-NAME) is a non-selective inhibitor of NOS. We aimed to evaluate whether L-NAME treatment had protective effects against oxidative stress in rats intragastrically fed with ethanol during a 4 wk-period. Thirty-six male Wistar rats were divided into 3 equal groups: group 1 (control group-isocaloric dextrose was given), group 2 (6 g/kg.d ethanol-induced group) and group 3 (both ethanol 6 g/kg.d and L-NAME 500 mg/L in drinking water-given group). Animals were sacrificed at the end of 4 wk-experimental period, and intracardiac blood and liver tissues were obtained. Biochemical measurements were performed both in plasma and in homogenized liver tissues. Alanine amino transferase (ALT), aspartate amino transferase (AST), malondialdehyde (MDA), NO, superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) levels were measured by spectrophotometry. ALT and AST in group 2 (62 U/L and 128 U/L, respectively) were higher than those in group 1 (24 U/L and 38 U/L) and group 3 (37 U/L and 81 U/L) (P<0.001 for both). Plasma and tissue levels of MDA in group 2 (4.66 micromol/L and 0.55 nmol/mg protein) were higher than in group 1 (2.65 micromol/L and 0.34 nmol/mg protein) and group 3 (3.43 micromol/L and 0.36 nmol/mg protein) (P<0.001 for both). Plasma and liver tissue levels of NO in group 2 (54.67 micromol/L and 586.50 nmol/mg protein) were higher than in group 1 (34.67 micromol/L and 435.33 nmol/mg protein) and group 3 (27.50 micromol/L and 412.75 nmol/mg protein) (P<0.001 for both). Plasma and liver tissue SOD activities in group 2 (15.25 U/mL and 5.38 U/ mg protein, respectively) were lower than in group 1 (20.00 U/mL and 8.13 U/ mg protein) and group 3 (19.00 U/mL and 6.93 U/ mg protein) (P<0.001 for both). Plasma and liver tissue CAT activities in group 2 (145 U/mL and 37 U/ mg protein, respectively) were lower than in group 1 (176 U/mL and 73 U/mg protein) and group 3 (167 U/mL and 61 U/mg protein) (P<0.001 for both). Meanwhile, erythrocytes and liver tissue levels of GSH in group 2 (4.12 mg/g Hb and 5.38 nmol/mg protein, respectively) were lower than in group 1 (5.52 mg/g Hb and 4.49 nmol/mg protein) and group 3 (5.64 mg/g Hb and 4.18 nmol/mg protein) (P<0.001 for both). Our findings show that L-NAME may produce a restorative effect on ethanol-induced liver damage via decreasing oxidative stress and increasing antioxidant status.