Rat Kidney Enzyme Research Articles

Chemical characterization of Hymenocardia acida stem bark extract and modulation of selected enzymes in Kidney and Heart of Wistar rats

Hymenocardia acida tul leaf and stem bark are used in treatment of several diseases in Africa. We examined the chemical constituents of the stem bark extract and its effects on some antioxidant indices and esterases in Wistar rats. Hymenocardia acida stem bark extract (HASBE) was obtained by Soxhlet extraction using methanol, followed by Atomic Absorption Spectroscopy (AAS), Fourier-Transform Infrared (FT-IR) spectroscopy, ultraviolet (UV) spectroscopy, High-performance liquid chromatography (HPLC) and Gas Chromatography-Flame ionization detection (GC-FID). Forty-eight male Wistar rats were assigned into eight groups (6 rats each), and administered orally with normal saline (Control), 50, 100, 150, 200, 250, 300, 350 mg/kg of HASBE twice per week for eight weeks. The rats were sacrificed under chloroform anesthesia, and kidneys and heart were excised, and processed into homogenates. Superoxide dismutase (SOD), catalase, lipid peroxidation (LPO), glutathione peroxidase (GPx), acetylcholinesterase (AChE) and carboxylesterases (CES) were determined spectrophotometrically. The AAS of HASBE detected Cobalt, Copper, Zinc, Iron, Nickel, Chromium, Manganese and Magnesium. The FT-IR shows four peaks as 2961.4, 2926.0, 1056.7 and 1034.3 cm-1, while UV shows absorbance between 250 nm and 650 nm. The HPLC identified orientin, β-sitosterol, rutin and betulinic acid, while GC-FID identified rutin, orientin, stigmasterol, hymenocardine and homopterocarpin as prominent compounds. The SOD significantly (p < 0.05) reduced in kidneys, while catalase was elevated in kidney and heart, with an increase in LPO level only in heart, relative to controls. The GPx, AChE and CE activities in kidneys were increased by HASBE, whereas, CE activity was lowered in heart. This study has demonstrated that Hymenocardia acida stem bark extract majorly contains iron, nickel, orientin, rutin, stigmasterol, hymenocardine, β-sitosterol, homopterocarpin and betulinic acid, and could possibly modulate the activities of antioxidant and esterase enzymes in kidney and heart of Wistar rats.

Sexual Dimorphism in the Expression of Cytochrome P450 Enzymes in Rat Heart, Liver, Kidney, Lung, Brain, and Small Intestine.

Cytochrome P450 (P450) enzymes are monooxygenases that are expressed hepatically and extrahepatically and play an essential role in xenobiotic metabolism. Substantial scientific evidence indicates sex-specific differences between males and females in disease patterns and drug responses, which could be attributed, even partly, to differences in the expression and/or activity levels of P450 enzymes in different organs. In this study, we compared the mRNA and protein expression of P450 enzymes in different organs of male and female Sprague-Dawley rats by real-time polymerase chain reaction and western blot techniques. We found significant sex- and organ-specific differences in several enzymes. Hepatic Cyp2c11, Cyp2c13, and Cyp4a2 showed male-specific expression, whereas Cyp2c12 showed female-specific expression. Cyp2e1 and Cyp4f enzymes demonstrated higher expression in the female heart and kidneys compared with males; however, they showed no significant sexual dimorphism in the liver. Male rats showed higher hepatic and renal Cyp1b1 levels. All assessed enzymes were found in the liver, but some were not expressed in other organs. At the protein expression level, CYP1A2, CYP3A, and CYP4A1 demonstrated higher expression levels in the females in several organs, including the liver. Elucidating sex-specific differences in P450 enzyme levels could help better understand differences in disease pathogeneses and drug responses between males and females and thus improve treatment strategies. SIGNIFICANCE STATEMENT: This study characterized the differences in the mRNA and protein expression levels of different cytochrome P450 (P450) enzymes between male and female rats in the heart, liver, lung, kidney, brain, and small intestine. It demonstrated unique sex-specific differences in the different organs. This study is considered a big step towards elucidating sex-specific differences in P450 enzyme levels, which is largely important for achieving a better understanding of the differences between males and females in the disease's processes and treatment outcomes.

Oral Administration of Copper Chloride Damages DNA, Lowers Antioxidant Defense, Alters Metabolic Status, and Inhibits Membrane Bound Enzymes in Rat Kidney.

Copper (Cu) is a heavy metal that is widely used in industries and is also an essential micronutrient for living beings. However, excess Cu is toxic and human exposure to high levels of this metal results in numerous adverse health effects. We have investigated the effect of oral administration of copper chloride (CuCl2), a Cu(II) compound, on various parameters of oxidative stress, cellular metabolism, and DNA integrity in the rat kidney. This was done to delineate the molecular mechanism of Cu(II) toxicity. Adult male rats were randomly divided into five groups. Animals in four CuCl2-treated groups were separately administered single acute oral dose of CuCl2 at 5, 15, 30, and 40mg/kg body weight. Animals in the fifth group were not given CuCl2 and served as the control. All rats were sacrificed 24h after the dose of CuCl2 and their kidneys removed. CuCl2 administration led to significant alterations in enzymatic and non-enzymatic parameters of oxidative stress. It changed the activities of metabolic and membrane bound enzymes and also decreased the activities of brush border membrane enzymes. CuCl2 treatment dose-dependently enhanced DNA damage and DNA-protein crosslinking in renal cells, when compared to the control group. The administration of CuCl2 also resulted in marked morphological changes in the kidney, with more prominent alterations at higher doses of CuCl2. These results clearly show that CuCl2 impairs the antioxidant defense system resulting in oxidative damage to the kidney.

Effect of therapeutic hypothermia against renal injury in a rat model of asphyxial cardiac arrest: Α focus on the survival rate, pathophysiology and antioxidant enzymes.

Although multi-organ dysfunction is associated with the survival rate following cardiac arrest (CA), the majority of studies to date have focused on hearts and brains, and few studies have considered renal failure. The objective of the present study, therefore, was to examine the effects of therapeutic hypothermia on the survival rate, pathophysiology and antioxidant enzymes in rat kidneys following asphyxial CA. Rats were sacrificed one day following CA. The survival rate, which was estimated using Kaplan-Meier analysis, was 42.9% one day following CA. However, hypothermia, which was induced following CA, significantly increased the survival rate (71.4%). In normothermia rats with CA, the serum blood urea nitrogen level was significantly increased one day post-CA. In addition, the serum creatinine level was significantly increased one day post-CA. However, in CA rats exposed to hypothermia, the levels of urea nitrogen and creatinine significantly decreased following CA. Histochemical staining revealed a significant temporal increase in renal injury after the normothermia group was subjected to CA. However, renal injury was significantly decreased in the hypothermia group. Immunohistochemical analysis of the kidney revealed a significant decrease in antioxidant enzymes (copper-zinc superoxide dismutase, manganese superoxide dismutase, glutathione peroxidase and catalase) with time in the normothermia group. However, in the hypothermia group, these enzymes were significantly elevated following CA. Collectively, the results revealed that renal dysfunction following asphyxial CA was strongly associated with the early survival rate and therapeutic hypothermia reduced renal injury via effective antioxidant mechanisms.

Fish Oil Supplementation Ameliorates Long Term Cisplatin Treatment Induced Toxicity and Oxidative Damage in Rat Kidney

Background: Cisplatin (CP) is an effective chemotherapeutic agent widely used against various malignancies. However, CP exerts several side effects through oxidative stress by generating reactive oxygen species (ROS). The major side effect of CP chemotherapy is dose dependent nephrotoxicity that limits its clinical usage. We have previously shown that fish oil (FO) supplementation ameliorated gentamicin induced nephrotoxicity as evident by increase in the activities of brush border membrane (BBM) and free radical scavenging enzymes in rat kidney. In the present study we aimed to investigate whether fish oil (FO) can prevent the long term CP-induced nephrotoxic effects. Methods: Rats were divided into four groups viz. control, CP, CPF and FO alone group. Rats in the groups CPF and FO were fed experimental diet prior to and simultaneously with or without CP treatment (3 mg/kg bwt i.p) respectively. Results: FO supplementation to CP treated significantly mitigated the CP induced increase in serum creatinine (Scr) and blood urea nitrogen (BUN) and decrease in the activities of brush border membrane (BBM) enzymes in renal cortical and medullary homogenates as well as in isolated BBM vesicles (BBMV). FO consumption also reduced CP induced alterations in the activities of carbohydrate metabolism enzymes and in the enzymatic and non-enzymatic antioxidant parameters. Conclusion: These results suggest that FO have potential to abrogate CP induced nephrotoxicity and can be a useful adjuvant in long term CP chemotherapy without any side effects. Moreover, the inherent anti-cancer properties of FO with CP treatment may exhibits synergistic beneficial effects.

Protective effect of magnesium lactate gluconate and Garcinia cambogia fruit extract in experimentally induced renal calculi in rats

Antiurolithiatic acitivity of magnesium lactate gluconate (MLG) and aqueous extract of Garcinia cambogia (GC) fruit was studied. Methods: Study was performed during December 2016 to April 2017. Urolithiasis was induced in male Wistar rats by administration of 0.75 % v/v ethylene glycol for 21 days. From 8th day onwards, intervention with MLG (200 and 400 mg/kg b.w.) and GC (100 and 200 mg/kg b.w.) was started. At the end of treatment period, biochemical parameters affecting renal stone formation were estimated in serum, urine, kidney homogenate and histopathology of harvested kidneys was performed. Results: From in vivo evaluation, it was observed that MLG 400 mg/kg b.w., GC 100 mg/kg b.w. and GC 200 mg/kg b.w. significantly reduced nitrogenous waste products in serum (blood urea nitrogen, creatinine, uric acid) as well as calculogenic promoters in urine (phosphate, oxalate) and kidney homogenate (calcium, phosphate, oxalate) when compared to disease control animals. The MLG 200 and MLG 400 were ineffective in restoring superoxide dismutase (SOD) and catalase (CAT) enzyme activity whereas GC 100, GC 200 and Cystone® 400 mg/kg b.w. significantly elevated SOD and CAT enzymes in urolithiatic rat kidney. Conclusions: MLG and GC extract are capable of preventing calcium oxalate (CaOx) crystal formation and subsequent deposition in renal tubules. The principle mechanism underlying nephroprotective effect of test drugs might be attributed to their calcium ion cheating ability and CaOx crystallization inhibitory activity. It is further asserted that GC was more potent than MLG in overall kidney protection by virtue of its antioxidant potential.

Effects of antipsychotic drug administration on antioxidative defense enzymes in male rat kidney

ABSTRACTThe use of atypical antipsychotic drugs (APD) was reported to be associated with adverse effects on the kidneys. Thus, the aim of this study was to examine whether APD exerted their adverse effects by interfering with the renal antioxidant defense system. Male 3-mo-old Wistar rats were treated for 28 d with ziprasidone (ZIP), clozapine (CLO), or sertindole (SER) using a daily dose recommended for antipsychotic drug therapy. The expression and activities of antioxidant enzymes superoxide dismutase (SOD) type 1 and type 2, catalase (CAT), glutathione reductase (GR), and glutathione S-transferases (GSTs) activity were measured in the kidneys. Changes in the kidneys were also evaluated histologically. Ziprasidone, CLO, and SER reduced renal SOD type 1 and type 2 activities. Decreased CAT activity was observed only in SER-treated rats. An inhibition in GR activity and increased activity of GST was found only after treatment with CLO. Histological analysis showed dilatation of proximal tubules in kidneys with all three drugs. In conclusion, data indicate that redox disturbances may contribute to renal morphologic alterations in proximal tubules in rats treated with all APD.

Gene Expression and Activities of Antioxidant Enzymes in Kidneys of Rats Intoxicated with Aflatoxin B1

Gene Expression and Activities of Antioxidant Enzymes in Kidneys of Rats Intoxicated with Aflatoxin B1

Effects of treatment with the anti-parasitic drug diminazene aceturate on antioxidant enzymes in rat liver and kidney.

Diminazene aceturate (DA) is the active component of some trypanocidal drugs used for the treatment of animals infected with trypanosomosis and babesiosis. Residues of DA may cause hepatotoxic and nephrotoxic effects. Therefore, the purpose of this study was to investigate the occurrence of oxidative stress, i.e., changes in the antioxidant defense system of rats treated with a single dose of 3.5 mg kg(-1) of DA. All treatments were intramuscularly administered, and evaluations were performed on days 7 and 21 post-treatment (PT). Liver and kidney samples were collected and evaluated by histopathology and oxidative stress parameters (thiobarbituric acid-reactive species, catalase, superoxide dismutase, carbonyl, non-protein thiols, and reduced glutathione). Finally, blood was collected to determine seric DA concentration. Superoxide dismutase (SOD) and catalase (CAT) activities in liver and kidney of rats were dramatically inhibited (p < 0.05) compared to the control group on day 21 PT. This difference is related to the concomitant increase (p < 0.05) in malondialdehyde (MDA) content, which was identified by an increase in thiobarbituric acid-reactive species (TBARS) levels. The carbonyl levels did not differ between groups (p > 0.05). Both non-protein thiols (NPSH) and glutathione (GSH) levels in liver and kidney decreased (p < 0.05) on day 21 PT. Chromatographic analyses showed lower levels of DA on day 21 PT compared to day 7 PT. A negative correlation was observed between DA concentration in serum and lipid peroxidation in liver and kidney tissues on 21 days PT. Histopathology revealed vacuolar degeneration in liver and kidney samples on day 21 PT. Our findings indicate that DA could cause oxidative damage to liver and kidney of rats.

Aspartame Induce Modification in Membrane Bound and Antioxidant Enzymes in Liver and Kidney of Wistar Albino Rats

Aspartame is rapidly and completely metabolized in humans and experimental animals to aspartic acid (40%), phenylalanine (50%) and methanol (10%). Methanol, a toxic metabolite is primarily metabolized by oxidation to formaldehyde and then to formate these processes are accompanied by the formation of superoxide anion and hydrogen peroxide. This study focuses on whether the oral administration of aspartame (40 mg/kg.bw/day) for 15-days and 30- days, has any effect on membrane bound ATPase, ‘Marker’ enzymes (ALP, ALT, ACP, AST &LDH) some selective liver and kidney function parameter and antioxidant status in liver and kidney of rats. To mimic human methanol metabolism, folate deficient rats were used. After 15-days of aspartame administration, there was no change observed in kidney, however liver showed a significant decrease in membrane bound ATPase enzyme (Na + /k + , Mg 2+ & Ca 2+ ), marker enzymes (ALP, ALT, AST & LDH) and significant increase in marker enzymes (ACP), antioxidant enzyme level. However, after aspartame administration for30-days, liver and kidney both showed a significant decrease in membrane bound ATPase enzyme( Na + k + , Mg 2+ & Ca 2+ ), marker enzymes (ALP, ALT, AST & LDH) and decrease of antioxidant enzyme level in liver and significant increase in marker enzymes (ACP) and antioxidant enzyme level in kidney. This study concludes that oral administration of aspartame (40mg/kg.bw/day) for longer duration may induce oxidative stress by aspartame metabolite methanol and can damage both liver and kidney. The liver was found to be more sensitive than kidney to oxidative stress, as also reflected by the histology of liver and kidney.

Effect of Aqueous Extract from Phaseolus vulgaris Pods on Lipid Peroxidation and Antioxidant Enzymes Activity in The Liver and Kidney of Diabetic Rats

Suppressive effect of Ginkgo biloba extract (EGb 761) on topsin induced ovarian toxicity and oxidative stress in albino ratsSaber A. Sakr,Hoda A. Mahran, Asmaa M. Abdel-Maksoud

Influence of testosterone on phase-II enzymes in liver and kidney of benzene-treated rats

Testosterone manifests some protective effects against benzene-induced toxicity in rats. However, mechanism of protection remains to be established. Data showed that testosterone modulates conjugation of reactive metabolites of benzene by influencing phase-II enzymes viz. glutathione-S-transferase, glutathione peroxidase, and catalase in both liver and kidney. These observations are supported by the opposite results obtained in castrated rats. It is postulated that testosterone decreases the formation of reactive oxygen species resulting into an increase in phase-II enzymes. Enhanced activity of these antioxidant enzymes is responsible for DNA strand repair as demonstrated by short comet tail length in liver and kidney of benzene and testosterone treated rats. Castration alters benzene pharmacokinetics by influencing these enzymes, a response which may be abolished by testosterone supplementation.

Modulatory effects of diosgenin on attenuating the key enzymes activities of carbohydrate metabolism and glycogen content in streptozotocin-induced diabetic rats.

Fenugreek and its active compound diosgenin are ancient herbal medicines recommended by the World Health Organization. In this study, the effect of diosgenin on changes in carbohydrate metabolic enzymes and glycogen content in muscle and kidneys of streptozotocin-induced diabetes rats were evaluated. Diabetes was induced in male albino Wistar rats by intraperitoneal administration of streptozotocin. The diosgenin at different doses (15, 30 and 60 mg/kg body weight) was administered orally to normal and streptozotocin-diabetic rats for 45 days. Streptozotocin intoxication led to a significant increase (p<0.05) in blood glucose and a decrease in insulin levels. The carbohydrate metabolic enzymes and glycogen content were also altered. The daily oral administration of diosgenin at different doses (15, 30 and 60 mg/kg body weight) to diabetic rats for 45 days resulted a significant (p<0.05) decline in blood glucose level and a significant increase in plasma insulin level. The altered activities of carbohydrate metabolic key enzymes in muscle and kidneys of diabetic rats were significantly (p<0.05) reverted to near normal level by the administration of diosgenin. The obtained results were compared with glibenclamide, a standard oral hypoglycemia drug. The modulatory effects of diosgenin on attenuating the activities of carbohydrate metabolic enzymes afford a promise for persistent use for the treatment of diabetes in the future, even though clinical studies to evaluate this possibility may be warranted.

Активність антиоксидантних ензимів нирок

Salts of heavy metals are excreted by the kidneys and, as pro-oxidants, stimulate the processes of free radical oxidation. Mercury ions are accumulated in the kidneys. So the study of the features of antioxidant enzymes adaptive response of different kidney layers in response to mercury dichloride is important. Catalase and glytathionperoxidase activity within rat kidneys 72 hours after mercury dichloride intoxication in the ratio of 5 ml per 1 kg of the animal weight was studied. It was important to reveal the influence of the mercury salts on rat kidney antioxidative system. Decreasing glytathionperoxidase activity in cortical and cerebral substances and renal papillae were accompanied by increased contents of oxidative modified proteins and lipids and morphological changes in renal tissue under salt and water loading after mercury dichloride poisoning. The results obtained evidence for the inhibition of antioxidative protection of enzymes in rat kidneys under the mercury dichloride effect.

Activity and genetic expression of antioxidant enzymes in liver and kidney of rats exposed to a modified clay and its migration extract

Activity and genetic expression of antioxidant enzymes in liver and kidney of rats exposed to a modified clay and its migration extract

Effects of Aqueous Extract of Nauclea latifolia Stem on Lipid Profile and Some Enzymes of Rat Liver and Kidney

Effects of Aqueous Extract of Nauclea latifolia Stem on Lipid Profile and Some Enzymes of Rat Liver and Kidney

Changes in expression profiles of antioxidant enzymes in diabetic rat kidneys

In diabetes mellitus, increased formation of reactive oxygen species due to high level of glucose in both blood plasma and tissues creates oxidative stress and damages the tissues. Antioxidants together with the antioxidant enzymes are very important in order to protect the cells against oxidative damage. Differential expressions of both mRNA and proteins of major antioxidant enzymes in streptozotocin-induced diabetic rat kidneys were measured with the help of real-time polymerase chain reaction and western blot analysis, respectively. Furthermore, effects of two strong antioxidants α-lipoic acid, vitamin C and their combination on the regulation of both expressions and the activities of antioxidant enzymes were also studied. In diabetic rat kidney tissue, both catalase and glutathione peroxidase activities were reduced (although mRNA expression for both was greatly increased) when compared with controls. No significant change was observed in superoxide dismutase (SOD) activity. Alpha-lipoic acid increased catalase activity towards the control values. Combined administration of alpha-lipoic acid and vitamin C increased the activities of both catalase and SOD, demonstrating a posttranslational effect. Glutathione concentrations were decreased in diabetic kidney; alpha-lipoic acid treatment partially restored the glutathione levels. All data showed the importance of post-transcriptional and translational regulation of the antioxidant enzyme activities against oxidative stress that is associated with diabetes.

Effects of chitosan oligosaccharides on drug-metabolizing enzymes in rat liver and kidneys

To investigate the effect of chitosan oligosaccharides (COS) on drug-metabolizing enzymes in rat liver and kidneys, male Spraque–Dawley rats were fed a diet containing 1% or 3% COS for 5weeks. The activities of cytochrome P450 (CYP) enzymes, UDP-glucurosyltransferase (UGT) and glutathione S-transferase (GST) in the liver and kidneys were determined. Significant decreases in microsomal CYP3A-catalyzed testosterone 6β-hydroxylation, CYP2C-catalyzed diclofenac 4-hydroxylation, and CYP4A-catalyzed lauric acid 12-hydroxylation in the liver of rats fed the COS diets were observed compared with those rats fed the control diet. Immunoblot analyses of CYP proteins showed the same trend as with enzyme activities. Increased glutathione content in liver was found in rats fed the 1% COS diet. Increased hepatic NADPH: quinone oxidoreductase 1 (NQO1) activity was found in rats fed the COS diets. In kidneys, COS had little or no effect on CYP enzyme activities. However, increased GST activity was observed in rats fed the COS diets. Moreover, a higher UGT activity was found in rats fed the 1% COS diet. Our results indicate that COS may suppress hepatic CYP enzymes and induce phase II detoxifying reactions in the liver and kidneys of rats.

Effect of Pimpinellatirupatiensison Oxidative Enzymes in STZ-induced Diabetic Rat Kidney.

The present study was aimed to evaluate the therapeutic potential of Pimpinellatirupatiensis(Pt) by assaying the activities of selective mitochondrial enzymes in streptozotocin induced diabetic rats. Diabetic rats showed a significant (p < 0.01) reduction in the activities of oxidative enzymes Succinate dehydrogenase (SDH), Malate dehydrogenase (MDH), Glutamate dehydrogenase (GDH) and isocitrate dehydrogenase (ICDH). Lactate dehydrogenase (LDH) activity was significantly (p < 0.01) increased in diabetic rats. The daily oral treatment of Pimpinellatirupatiensisethyl alcohol extract (750 mg/kg body weight/day) to diabetic rats for 30 days reversed the above changes in a significant (p < 0.01) manner. From our observations, we conclude that administration of Pt altered the activities of oxidative enzymes, thereby suggesting its role in mitochondrial energy production. The obtained results were compared with Glibenclamide, a standard anti diabetic drug. Thus, the modulatory effects of Pt on altering these enzymes activities afford a promise for widespread use for treatment of diabetes in the future.

Influence of rat strains and/or severity of hyperglycemia on systolic blood pressure and antioxidant enzymes in kidney of rats with hypertension and/or diabetes: Role of honey

Influence of rat strains and/or severity of hyperglycemia on systolic blood pressure and antioxidant enzymes in kidney of rats with hypertension and/or diabetes: Role of honey