Antioxidative effects of pumpkin seed (Cucurbita pepo) protein isolate in CCl4-induced liver injury in low-protein fed rats

The effects of pumpkin seed (Cucurbita pepo) protein isolate on the plasma activity levels of catalase (CA), superoxide dismutase (SOD), glutathione peroxidase (GSHpx) and total antioxidant capacity (TAC) as well as glucose-6-phosphatase (G6Pase) in liver homogenates and lipid peroxidation (LPO-malondialdehyde-MDA) levels in liver homogenates and liver microsomal fractions against carbon tetrachloride (CCl(4))-induced acute liver injury in low-protein fed Sprague-Dawley rats (Rattus norvegicus) were investigated. A group of male Sprague-Dawley rats maintained on a low-protein diet for 5 days were divided into three subgroups. Two subgroups were injected with carbon tetrachloride and the other group with an equivalent amount of olive oil. Two hours after CCl(4) intoxication one of the two subgroups was administered with pumpkin seed protein isolate and thereafter switched onto a 20% pumpkin seed protein isolate diet. The other two groups of rats were maintained on the low-protein diet for the duration of the investigation. Groups of rats from the different subgroups were killed at 24, 48 and 72 h after their respective treatments. After 5 days on the low-protein diet the activity levels of all the enzymes as well as antioxidant levels were significantly lower than their counterparts on a normal balanced diet. However, a low-protein diet resulted in significantly increased levels of lipid peroxidation. The CCl(4) intoxicated rats responded in a similar way, regarding all the variables investigated, to their counterparts on a low-protein diet. The administration of pumpkin seed protein isolate after CCl(4) intoxication resulted in significantly increased levels of all the variables investigated, with the exception of the lipid peroxidation levels which were significantly decreased. From the results of the present study it is concluded that pumpkin seed protein isolate administration was effective in alleviating the detrimental effects associated with protein malnutrition and CCl(4) intoxication. It is therefore apparent that pumpkin seed protein isolate has components that have antiperoxidative properties.     

齐墩果酸对大鼠实验性肝损伤作用机理的研究

OLA(50、100、150mg/kg)灌胃,一天一次,连续6天,呈剂量依赖性地降低CCl_4染毒大鼠肝MDA含量和SGPT活性,减少肝TG蓄积,提高肝GSH含量。在100、150mg/kg时,明显提高肝5＇-NT活性。提示,OLA抗肝损伤的作用机理可能与提高肝GSH含量而抑制肝细胞脂质过氧化反应有关。     

Evaluation of the effectiveness of Rosmarinus officinalis (Lamiaceae) in the alleviation of carbon tetrachloride-induced acute hepatotoxicity in the rat

The effect of oral administration of Rosmarinus officinalis L. (Lamiaceae) on CCl(4)-induced acute liver injury was investigated. Rats were daily treated with the plant extract at a dose of 200 mg/kg corresponding to 6.04 mg/kg of carnosol as determined by reverse phase high-performance liquid chromatography. The treatment was initiated 1 h after CCl(4) administration and Rosmarinus officinalis fully prevented CCl(4) effect on hepatic lipid peroxidation after 24 h of CCl(4) administration. The increase in bilirubin level and alanine aminotransferase activity in plasma induced by CCl(4) was completely normalized by Rosmarinus officinalis. The treatment also produced a significant recovery of CCl(4)-induced decrease in liver glycogen content. CCl(4) did not modify the activity of liver cytosolic glutathione S-transferase (GST) compared with that of control groups. However, Rosmarinus officinalis increased liver cytosolic GST activity and produced an additional increment in plasma GST activity in rats treated with CCl(4). Histological evaluation showed that Rosmarinus officinalis partially prevented CCl(4)-induced inflammation, necrosis and vacuolation. Rosmarinus officinalis might exert a dual effect on CCl(4)-induced acute liver injury, acting as an antioxidant and improving GST-dependent detoxification systems.     

Effects of black tea extract on carbon tetrachloride-induced lipid peroxidation in liver, kidneys, and testes of rats

Previous studies have shown that green tea and black tea have antioxidant effects and chemopreventive activity against chronic disease including some forms of cancer. We have, therefore, examined the effects of an aqueous extract of black tea against carbon tetrachloride-induced lipid peroxidation as determined by the formation of thiobarbituric acid reactive substances in liver, kidneys and testes of rats. A 0.7% black tea extract was used which contained 2 mg of black tea extract solids per mL. Black tea was administered as drinking water for 3, 6, 9 and 12 months before and during carbon tetrachloride (CCl(4)) treatment in female and male rats. Rats were treated with a single oral dose of CCl(4) 1.0 mL/kg. All rats were killed 24 h after CCl(4) treatment. All animals were dosed with CCl(4) at the end of the 3, 6, 9, and 12 month of treatment. Black tea treatment for 75 days produced a decrease in CCl(4)-induced hepatic lipid peroxidation but significant decreases in thiobarbituric acid reactive substances occurred 3 months after treatment in both female and male rats. In liver and kidneys, black tea alone increased lipid peroxidation by 30%-50% in female and male rats. However, black tea decreased CCl(4)-induced lipid peroxidation in liver of female and male rats by approximately 49% and 37%, respectively. Black tea decreased CCl(4)-induced lipid peroxidation in testes by approximately 37% at a dose of 1.0 mL CCl(4)/kg. These results suggest that the protective effects of black tea against CCl(4)-induced lipid peroxidation in liver, kidneys and testes is due at least partly to its antioxidant properties, scavenging CCl(4)-associated free radicals.     

Hepatoprotective effect of the aqueous extract of Flemingia macrophylla on carbon tetrachloride-induced acute hepatotoxicity in rats through anti-oxidative activities

This study investigated the protective effect of the aqueous extract of Flemingia macrophylla (AFM) against hepatic injury induced by CCl(4). Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected as biomarkers in the blood to indicate hepatic injury. Product of lipid peroxidation (MDA), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and reduced glutathione (GSH) contents were evaluated for oxidative stress in hepatic injury. Moreover, histopathological observation was assayed for the degree of hepatic injury. After oral administration of AFM, 0.5 g/kg and 1.0 g/kg doses significantly decreased ALT and AST, attenuated the histopathology of hepatic injury, ameliorated oxidative stress in hepatic tissue, and increased the activities of CAT, SOD and GSH-Px. The hepatoprotective effect of daidzein and genistein were consistent to that of AFM. This study demonstrated for the first time that AFM has hepatoprotective effect on acute liver injuries induced by CCl(4), and the results suggested that the effect of AFM against CCl(4)-induced liver damage was related to antioxidant properties.     

Ameliorative effect of Silene aprica on liver injuries induced by carbon tetrachloride and acetaminophen

The effect of oral administration of a 50% ethanol extract of Silene aprica (SA) on acute liver injury was examined in rats intoxicated with carbon tetrachloride (CCl4) and acetaminophen. The results indicated that SA protected the liver from CCl4- and acetaminophen-induced injury as judged by morphological and biochemical observations. An increase in both lipid peroxidation (LPO) and triglyceride concentrations occurred in the liver with CCl4 injection, SA administration significantly reduced these changes.     

Ameliorative effects of pycnogenol on carbon tetrachloride-induced hepatic oxidative damage in rats

This study evaluated the putative antioxidant activity of Pycnogenol (PYC) against CCl4-induced hepatic oxidative damage in Sprague-Dawley rats. A single oral dose of CCl4 (1.25 mL/kg) produced significantly increased levels of serum aminotransferase (AST) and alanine aminotransferase (ALT) activities. In addition, increased malondialdehyde (MDA) concentration, reduced glutathione (GSH) content, and decreased catalase, superoxide dismutase (SOD) and glutathione-S-transferase (GST) activities were observed in the hepatic tissues. However, concomitant administration with PYC (10 or 20 mg/kg) significantly improved CCl4-induced hepatic injury, as evidenced by the decline of serum AST and ALT activities in a dose dependent manner. Moreover, PYC reduced MDA concentration and increased GSH levels and catalase, SOD and GST activities in hepatic tissues, indicating that concomitant administration with PYC efficiently prevent the CCl4-induced oxidative damage in rats. The free radical scavenging assay showed that PYC has a dose-dependent scavenging activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radicals. These results indicate that PYC has an antioxidant effect against CCl4-induced hepatic oxidative damage and is useful as a hepatoprotective agent against various liver diseases induced by oxidative stress.     

Ameliorative effect of an urinary preparation on acetaminophen and D-galactosamine induced hepatotoxicity in rats.

The effect of oral administration of a preparation of human urine (PHU) on acute liver injury was examined in rats intoxicated with acetaminophen and D-galactosamine. The results indicated that PHU protected the liver from acetaminophen and D-galactosamine-induced injury as judged by morphological and biochemical observation. An increase in lipid peroxide concentrations and decrease in protein concentrations occurred in the liver by D-galactosamine injection, PHU administration significantly prevented these changes.     

Hepatoprotective activity of Emblica officinalis and Chyavanaprash

Hepatoprotective activity of Emblica officinalis (EO) and Chyavanaprash (CHY) extracts were studied using carbon tetrachloride (CCl(4)) induced liver injury model in rats. EO and CHY extracts were found to inhibit the hepatotoxicity produced by acute and chronic CCl(4) administration as seen from the decreased levels of serum and liver lipid peroxides (LPO), glutamate-pyruvate transaminase (GPT), and alkaline phosphatase (ALP). Chronic CCl(4) administration was also found to produce liver fibrosis as seen from the increased levels of collagen-hydroxyproline and pathological analysis. EO and CHY extracts were found to reduce these elevated levels significantly, indicating that the extract could inhibit the induction of fibrosis in rats.     

柔肝降酶合剂对大鼠四氧化碳诱导肝损害的防治作用

目的：观察柔肝降酶合剂治疗四氯化碳（CCl4）所致大鼠急性肝损伤的效果。方法：SD 雄性大鼠60 只,随机分成6 组,分别为柔肝降酶合剂低、中、高3 个剂量组、葡醛内酯组、正常组及模型组,采用CCl4 腹腔注射致大鼠急性肝损伤模型,各中药组每日灌胃柔肝降酶合剂,正常组和模型组给予蒸馏水灌胃,葡醛内酯组灌胃葡醛内酯水溶液,于实验的第12 天,禁食16 澡后处死大鼠,检查肝组织病理学改变,测定大鼠血清丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）,肝组织匀浆测定大鼠肝组织超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）、还原型谷胱甘肽（GSH）、过氧化氢酶（CAT）及丙二醛（MDA）,RT-PCR 法检测大鼠肝脏血红素氧化酶（HO-1）mRNA 表达水平。结果：与模型组比较,柔肝降酶合剂各剂量组大鼠肝病理损害均不同程度减轻,柔肝降酶合剂各剂量组及葡醛内酯组血清ALT 及AST含量均明显降低（P＜0.05 或P＜0.01）;柔肝降酶合剂各剂量组、葡醛内酯组肝匀浆GSH、GSH-Px、CAT、SOD 含量明显升高（P＜0.05 或P＜0.01）,MDA 含量明显降低（P约0.05 或P约0.01）;各治疗组肝脏HO-1 mRNA 相对表达量明显升高（P＜0.05 或P＜0.01）。结论：柔肝降酶合剂防治CCl4 所致大鼠急性肝损伤疗效确切。     

In vitro and in vivo antioxidant activity of alfalfa (Medicago sativa L.) on carbon tetrachloride intoxicated rats

The present study was conducted to determine whether lyophilized aqueous extract of alfalfa, or Medicago sativa L. could exert antioxidant activity against carbon tetrachloride-induced oxidative stress and liver injury in rats. The hepatoprotective activity of alfalfa extract was determined by assessing the levels of serum transaminases, ALP, bilirubin and lipid profile. Further, the effect of the test substance on malondialdehyde (MDA), an end product of lipid peroxidation; antioxidant liver enzyme non-protein sulfhydryl (NP-SH); and total protein (TP) were also studied. Serum transaminase, ALP, bilirubin level, lipid profile and liver MDA were significantly elevated and the antioxidant status in liver NP-SH and TP contents were declined in animals treated with CCl (4) alone. Pretreatment with alfalfa and silymarin for three weeks prior to the administration of CCl (4) significantly prevented the increase in the serum levels of hepatic marker, LDL, VLDL levels enzymes and reduced oxidative stress indicated by elevated NP-SH and TP concentration. The histopathological examination of the livers also showed that the alfalfa extract reduced the incidence of liver lesions induced by CCl (4). The in vitro antioxidant assessment of alfalfa extract on DPPH and carotene-linoleic assays demonstrated a moderate antioxidant potential. Results suggest that the alfalfa extract possesses hepatoprotective and antioxidative stress properties possibly through its antioxidant phytochemical constituents and substantiates its use in various liver disorders as a hepatoprotector.     

秃疮花提取物对CCl4诱导的小鼠急性肝细胞微粒体损伤的影响

目的:探讨研究秃疮花提取物(DLE)对CCl4诱导的小鼠急性损伤肝细胞微粒体的影响。方法:小鼠灌胃给予不同剂量的DLE,连续2周。最后一次给药24h后CCl4造成急性肝损伤。制备肝细胞微粒体悬液,检测抗氧化酶SOD、CAT、GSH-Px的活性;测量肝细胞微粒体悬液中的MDA水平;观察NADPH-细胞色素C(P450)还原酶活性及ATPase活性的变化。结果:与模型组相比,不同剂量的DLE给药组肝细胞微粒体悬液中MDA含量显著减少;抗氧化酶SOD、GSH-Px活性显著增加;肝细胞微粒体NAD-PH-细胞色素C(P450)还原酶活性降低,Na+K+-ATPase和Ca2+-ATPase活性显著上升。结论:DLE对CCl4诱导的小鼠肝微粒体损伤具有很好的保护作用。其作用机理与DLE具抗氧化和自由基活性清除有关。     

Protective effects of Ginkgo biloba, Panax ginseng, and Schizandra chinensis extract on liver injury in rats

This study investigated the effects of the combined extracts of Ginkgo biloba, Panax ginseng, and Schizandra chinensis at different doses on hepatic antioxidant status and fibrosis in rats with carbon tetrachloride (CCl4)-induced liver injury. Male Sprague-Dawley rats (n = 8-12 per group) were divided into the control, CCl4, CCl4 + silymarin (0.35%), CCl4 + low-dose herbal extract (0.24% of Ginkgo biloba, Panax ginseng, and Schizandra chinensis extract at 1:1:1; LE), and CCl4 + high-dose herbal extract (1.20% of the same herbal extract; HE) groups. Silymarin or herbal extract was orally given to rats a week before chronic intraperitoneal injection with CCl4 for 6 weeks. The pathological results showed that herbal extract suppressed hepatic bile duct proliferation, and low-dose herbal extract inhibited liver fibrosis. Hepatic superoxide dismutase (SOD) activity was lower in the CCl4 group, but there was no difference in the silymarin or herbal extract treated groups compared to the control group. Hepatic catalase activity and the ratio of reduced to oxidized glutathione were significantly higher (p < 0.05) in the HE group than those in the CCl4 group. Silymarin and herbal extract reversed the impaired hepatic total antioxidant status (p < 0.05). Herbal extract partially reduced the elevated hepatic lipid peroxides. Hepatic transforming growth factor-beta1 (TGF-beta1) level decreased significantly (p < 0.05) in the LE group. Therefore, high-dose herbal extract improved hepatic antioxidant capacity through enhancing catalase activity and glutathione redox status, whereas low-dose herbal extract inhibited liver fibrosis through decreasing hepatic TGF-beta1 level in rats with CCl4-induced liver injury.     

Effects of fig latex on lipid peroxidation and CCl4-induced lipid peroxidation in rat liver

The oral and intraperitoneal effects of fig milk latex on lipid peroxidation and CCl4-induced lipid peroxidation in liver homogenates of female rats were investigated. Oral treatment had no effect, while i.p. administration produced a significant increase in hepatic lipid peroxidation. When the latex was given before CCl4 treatment; it produced no protective effect against CCL4-induced hepatotoxicity. Addition of the latex to the incubation mixture produced a dose-dependent increase in lipid autoxidation, while the chloroform and ether extracts of the latex, as well as heated latex, had no effect on hepatic lipid autoxidation.     

Protective effect of Aquilegia vulgaris (L.) on APAP-induced oxidative stress in rats

Rats pretreated with acetaminophen (N-acetyl-p-aminophenol, APAP) (600 mg/kg b.w., p.o.) were administered with ethanol and ethyl acetate extracts as well as with isocytisoside (100 mg/kg b.w., p.o.) obtained from Aquilegia vulgaris (L.) (Ranunculaceae) herb. The substances tested decreased enzymatic, non-enzymatic and uninduced microsomal lipid peroxidation (LPO) in the liver of rats treated with APAP by 18-48%. Activity of the antioxidant enzymes in the liver inhibited by APAP was increased in the majority of groups after administration of the substances tested: catalase (CAT) by 55%, glutathione peroxidase (GPx) by 50%, glutathione reductase (GR) by 35% and glutathione S-transferase (GST) by 60%. Hepatic glutathione level depleted by APAP was only slightly increased by the substances tested. The cytochrome P450 contents, and the activities of NADPH-cytochrome P450 reductase and two monooxygenases were not affected by the extracts and isocytisoside. It can be concluded that the protective ability of the substances tested in APAP-induced liver injury is mediated by amelioration of microsomal lipid peroxidation and restoring antioxidant enzymes activity. Inhibition of enzymes responsible for metabolic activation of APAP is not involved in this process.     

玉郎伞黄酮对四氯化碳诱导的大鼠肝纤维化的影响

目的:研究玉郎伞黄酮(YLSF)对四氯化碳(carbon tetrachloride,CCl4)所致大鼠肝纤维化的治疗作用,并探讨其作用机制.方法:将SD大鼠随机分成模型组及空白对照组(NC),模型组以50％ CCl4食用油溶液为诱导剂ig造模,NC组以生理盐水ig.将病理检查确认形成肝纤维化的SD大鼠,随机分成模型对照组(MC)和药物干预组.药物干预组分别以YLSF(20,40,80 mg·kg-1)及秋水仙碱片(0.20 mg·kg-1)ig;MC组以等剂量生理盐水ig.末次给药24 h后处死大鼠,采集血清及肝组织,检测大鼠血清中天冬氨酸转氨酶( AST)、丙氨酸转氨酶(ALT)活性,测定肝组织中超氧化物岐化酶(SOD)、丙二醛(MDA)、谷胱甘肽(GSH)和谷胱甘肽过氧化酶(GSH-Px),并观察肝组织病理学改变.结果:YLSF(20,40,80 mg·kg -1)各剂量组大鼠血清中AST和ALT分别为(207.85±101.72),(131.55 ±35.09),(129.98±37.21)U·L-1和(90.51±44.24),(47.79 ±16.11),(44.56±16.31)U·L-1;YLSF各剂量组大鼠肝组织中SOD,MDA,GSH-Px,GSH含量分别为(143.14±42.82),(146.53±31.98),(147.41±32.82) U· mg-1,(2.57±0.54),(2.19±0.57),(2.11±0.59)nmol·mg-1,( 463.55±271.07),(659.14±162.23),(752.08±200.70) nmol·mg-1,(5.06±1.09),(6.10 ±0.97),(6.89 ±0.98) nmol·mg-1.各剂量YLSF和阳性药能显著降低大鼠血清中AST(P ＜0.01)及肝组织MDA含量(P＜0.05或P＜0.01),并显著提高肝组织中SOD含量(P ＜0.05或P＜0.01);中、高剂量YLSF和阳性药可降低大鼠血清中ALT水平(P＜0.01),显著升高大鼠肝组织GSH含量(P＜0.01);中、高剂量YLSF能显著升高大鼠肝组织GSH-Px(P ＜0.01).各剂量YLSF及阳性药均能够减轻肝细胞损伤程度(P ＜0.05或P＜0.01).结论:玉郎伞黄酮对CCl4诱导的大鼠具有一定治疗作用,其机制可能与其清除自由基、抑制脂质过氧化有关系.     

Protective effect of Lygodium flexuosum (L.) Sw. extract against carbon tetrachloride-induced acute liver injury in rats

The hepatoprotective potential of Lygodium flexuosum (L.) Sw. was evaluated in male Wistar rats against carbon tetrachloride-induced liver damage in preventive and curative models. Toxic control and n-hexane extract-treated rats received a single dose of CCl4 (150 microL/100g, 1:1 in corn oil). Pre-treated rats were given n-hexane extracts at 200 and 100 mg/kg dose 48, 24 and 2 h prior to CCl4 administration. In post-treatment groups, rats were treated with n-hexane extract at a dose of 200 and 100 mg/kg, 2, 24 and 48 h after CCl4 intoxication. Rats pre-treated with Lygodium flexuosum remarkably prevented the elevation of serum AST, ALT, LDH and liver lipid peroxides in CCl4-treated rats. Rats treated with the extract after the establishment of CCl4 induced liver injury showed significant (p < or = 0.05) protection of liver as evidenced from normal AST, ALT, LDH and MDA levels. Hepatic glutathione levels were significantly (p < or = 0.05) increased by the treatment with the extracts in both the experimental groups. Histopathological changes induced by CCl4 were also significantly (p < or = 0.05) reduced by the extract treatment in preventive and curative groups. Phytochemical studies revealed the presence of saponins, triterpenes, sterols and bitter principles in Lygodium flexuosumn-hexane extract which could be responsible for the possible hepatoprotective action.     

Scavenging of reactive oxygen species by a urinary preparation

In the present paper, the antioxidant properties of a preparation of human urine (PHU) were evaluated by studying the ability of this drug to react with relevant biological oxidants such as superoxide anion radical (O2*-) and hydroxyl radical (OH*). In addition, its effect on lipid peroxidation was investigated in vitro and ex vivo. PHU is not a good scavenger of O2*-. However, it reacts rapidly with OH radicals with a second-order rate constant of 2.8 x 10(9)/M/sec. The studies on rat brain homogenates showed that PHU had an inhibitory effect, which was dependent on its concentration and the magnitude of lipid peroxidation. Ex vivo studies also showed that oral administration of PHU increased the antioxidant capacity of plasma from rats. The ability of PHU to scavenge free radicals suggests that this drug may be potentially useful in counteracting free radical-mediated diseases.     

乙酰胡椒乙胺对小鼠化学中毒性肝损伤的保护作用

 目的 研究乙酰胡椒乙胺(PAA)对小鼠化学中毒性肝损伤的保护作用。方法 小鼠ig PAA后ip四氯化碳(CCl₄)或D-氨基半乳糖(D-Gal),检测肝损害的几项指标和肝脏的病理损害。结果 PAA可明显降低CCl₄,D-Gal引起的ALT升高,防止肝糖原下降,明显抑制由CCl₄体外引起的大鼠肝微粒体脂质过氧化。结论 PAA对CCl₄及D-Gal引起的肝损伤有明显保护作用。