罗布麻茶黄酮提取物对小鼠CYP2E1的影响

目的 CYP2E1是肝脏中主要的药物代谢酶,在肝脏疾病发生及发展中发挥重要的作用。本实验分别通过体内、体外实验,研究罗布麻茶黄酮提取物对小鼠CYP2E1活性和表达的影响,为评价罗布麻茶对肝脏的保护作用和药物的联合使用提供理论依据。方法将30只昆明小鼠随机分为空白对照组、黄酮提取物低剂量组和高剂量组(50和100 mg/kg),连续灌胃给药10天。采用探针药物法测定小鼠肝脏微粒体中CYP2E1催化对硝基苯酚代谢的活力。采用Western Blot法检测肝脏微粒体中CYP2E1蛋白的表达情况。体外抑制实验中,考察不同浓度的黄酮提取物对CYP2E1的抑制作用,计算半数抑制浓度(IC_(50))值。结果灌服高剂量黄酮提取物(100 mg/kg)后,小鼠体内CYP2E1酶活性和表达量均显著降低,黄酮提取物对CYP2E1的IC_(50)值为128.4μg/mL。结论罗布麻茶黄酮提取物对小鼠CYP2E1有抑制作用。     

基于Nrf2/CYP2E1通路探讨刺梨多糖对酒精性肝损伤小鼠的保护机制

目的:探讨刺梨多糖（Rosa roxburghii polysaccharide,RP）对食用酒精诱导的酒精中毒小鼠肝脏的影响及保护机制。方法:采用热水浸提法制得纯度为64％的RP。将60只健康KM雄性小鼠分为空白组、模型组、阳性组、RP低、中、高剂量组,连续给药15d,于末次给药4h后,除空白组外,其余各组一次性灌胃16 mL/kg 50％的食用酒精建立酒精性肝损伤（alcoholic liver disease,ALD）小鼠模型;解剖后记录脏器质量,测定血清中谷草转氨酶（aspartate aminotransferase,AST）、谷丙转氨酶（alanine aminotransferase,ALT）、总胆固醇（total cholesterol,TC）、甘油三酯（triglyceride,TG）含量、测定肝脏氧化应激指标含量,核因子-红细胞2相关因子2（nuclear factor erythroid2-related factor 2,Nrf2）信号通路相关蛋白表达量及细胞色素P450 2E1（cytochrome P450 2E1,CYP2E1）蛋白表达量、并进行肝脏病理和肝细胞凋亡观察。结果:与模型组相比,RP各剂量组均能降低肝脏指数、血清中转氨酶AST、ALT,血脂指标TC、TG含量、肝脏丙二醛（Malonaldehyde,MDA）含量,提高谷胱甘肽（glutathione,GSH）、超氧化物歧化酶（superoxide dismutase,SOD）含量,上调Nrf2、血红素氧合酶1(heme oxygenase-1,HO-1)、超氧化物歧化酶1（superoxide dismutase-1,SOD-1）蛋白表达量,下调CYP2E1蛋白表达量,并改善肝脏病变和肝细胞凋亡情况。结论:RP对酒精性肝损伤小鼠起到良好的护肝效果,这可能与调控Nrf2信号通路起到抗氧化作用及调节脂质代谢有关。     

牡蛎寡肽对H2O2氧化损伤L02人肝细胞的保护作用

为了研究牡蛎寡肽对自由基清除作用及对人肝细胞L02细胞氧化损伤的保护作用,测定了牡蛎寡肽对羟自由基(·OH)的清除率,并建立过氧化氢(hydrogen peroxide,H_2O_2)氧化损伤人肝细胞L02的模型,研究牡蛎寡肽对氧化损伤L02细胞的存活率、活性氧(Reactive Oxygen Species,ROS)、抗氧化酶系(superoxide dismutase/SOD,glutathione/GSH)含量的影响。结果表明,牡蛎寡肽对羟自由基的IC_(50)为0.38 mg/m L。2 mg/m L牡蛎寡肽对于L02细胞的增殖率仍为123.98%。模型组SOD和GSH水平分别降低了40%、64.87%,而ROS增加了1倍。当添加不同剂量牡蛎寡肽后,中剂量组细胞中SOD含量几乎恢复到正常组水平,GSH活性也提高了118.89%,ROS水平降到模型组的62.43%,说明牡蛎寡肽对L02细胞无毒,能降低氧化损伤的L02细胞内的ROS水平,显著提高SOD和GSH含量,对细胞起到保护作用。因此,牡蛎寡肽对H2O2致氧化损伤的人肝L02细胞具有保护作用,可能是通过清除自由基,保护细胞内抗氧化酶系活性,抑制ROS的过多积累来实现。     

酪蛋白水解物分别与维生素A、维生素C和维生素E协同保护乙醇氧化损伤的肝细胞

酪蛋白水解物可以治疗和修复乙醇氧化损伤的肝细胞。本文的研究目的在于评价维生素A、维生素C和维生素E分别与酪蛋白水解物(Casein Hydrolysate,CH)协同保护乙醇氧化损伤的肝细胞。已有研究表明,酪蛋白水解物对肝细胞HHL-5没有明显的毒性,甚至有着良好的促进增殖的结果。结果表明,300 mmo L/L的乙醇对细胞有着明显的损伤作用;酪蛋白水解物显著提高了乙醇损伤肝细胞HHL-5的细胞存活率。分别在维生素A(0.0344 mg/m L)、维生素C(0.0881 mg/m L)和维生素E(0.1077 mg/m L)的浓度下与酪蛋白水解物协同作用显著提高了乙醇损伤细胞的细胞存活率。通过CCK-8法和培养基内LDH含量的测定,选取最佳CH的协同浓度。通过胞内丙二醛含量的测定和胞内ROS含量的测定进行再次验证。2 mg/m L的酪蛋白水解物单独作用乙醇氧化损伤的细胞有着一定的修复作用,且该浓度下分别与维生素A、维生素C和维生素E协同保护作用显著增强(p0.05)。     

银杏黄酮对慢性酒精所致大鼠氧化损伤保护作用的研究

目的 研究银杏黄酮对慢性酒精中毒大鼠氧化损伤的保护作用。 方法 大鼠酒精灌胃,银杏黄酮采用预防性给药的方法。于实验第30d、60d分别检测大鼠血清中谷丙转氨酶(Glutamic-Pyruvic Transaminase, ALT)、谷草转氨酶(Glutamic Oxaloacetic Transaminase, AST)活性;谷胱甘肽(Glutathione, GSH)、活性氧(Reactive Oxygen Species,ROS)以及丙二醛(Malondialdehyde, MDA)的含量。 结果 慢性酒精摄入30 d和60 d后,酒精组血清ROS、ALT、AST和MDA较对照组明显升高,差异有极显著性(p<0.01),GSH则显著性降低 (p<0.01);与此同时,实验第30 d和60 d,低、高剂量银杏黄酮能明显降低酒精所致的ROS、ALT、AST、MDA升高(p<0.01),第60 d,高剂量银杏黄酮能显著抑制酒精所致的GSH降低(p<0.01)。结论 银杏黄酮对酒精所致氧化损伤具有一定的保护作用。     

丁香提取物对人肝细胞H_2O_2氧化损伤的保护作用及其机制

研究丁香提取物对由H2O2引起的人肝细胞HL770氧化损伤的保护作用及其机制。采用H2O2诱导建立细胞氧化损伤模型,通过测定细胞抗氧化酶活性、丙二醛、过氧亚硝基阴离子等指标,分析探讨丁香提取物对细胞损伤的保护作用。结果显示丁香水提取物和乙醇提取物可以剂量依赖性地提高细胞内的SOD、CAT和GSH-Px酶活性,降低氧化产物过氧亚硝基阴离子和丙二醛含量。表明丁香提取物可通过提高HL7702细胞内抗氧化酶活性和降低氧化产物的含量减轻细胞受到的氧化损伤。     

朝鲜蓟叶水提取物对酒精诱导人肝癌细胞株HepG2损伤的影响

目的通过建立酒精损伤HepG2细胞模型,探讨朝鲜蓟叶水提取物对酒精诱导的HepG2细胞损伤的影响。方法以人肝癌细胞株HpeG2为实验材料,采用酒精加入细胞培养液培养HepG2细胞,建立酒精损伤HepG2细胞模型,以MTT(3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide)法筛选酒精处理浓度、时间以及朝鲜蓟叶水提取物最佳作用浓度,通过检测各组细胞培养基中谷丙转氨酶(alanine aminotran-sferase,ALT)、谷草转氨酶(aspartate transaminase,AST)和细胞内超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)、甘油三酯(triglyceride,TG)的活性,来评价朝鲜蓟叶水提取物对酒精诱导的HepG2肝细胞损伤的影响。结果分别以0.6%、1.2%、2.4%的酒精浓度联合2.5、5、10μg/mL朝鲜蓟叶水提取物培养HepG2细胞48 h。模型组随着酒精浓度增高,细胞的AST、ALT、TG、MDA水平明显升高,SOD活性显著下降。处理组随着朝鲜蓟叶水提取物使用浓度的增加,细胞中AST、ALT、TG、MDA水平与模型组相比明显下降,SOD活性升高,对HepG2细胞的保护作用增强。结论朝鲜蓟叶水提取物在一定程度上能保护酒精受损的HepG2细胞,具有深入研究和开发的潜能。     

三种黄酮醇化合物对H2O2 和CCl4 诱导的人肝细胞体外损伤的保护作用比较

黄酮醇是一类广泛存在于植物界中的多酚物质,并具有许多生理作用,在食品中具有较大的应用潜力。通过一些生化实验方法,研究了3 种黄酮醇(山奈素、槲皮素、杨梅黄酮)对由H2O2 和CCl4 诱导的人肝细胞(HL-7702细胞系)体外氧化损伤的保护作用,并研究了3 种黄酮醇的化学结构与其细胞保护作用之间的关系。结果表明,3种黄酮醇(20、40、60μmol/L)与肝细胞预先作用30min,再进行诱导损伤,可以提高肝细胞的细胞活率和还原型谷胱甘肽(GSH)含量水平,降低乳酸脱氢酶(LDH)渗出率和丙二醛(MDA)生成量。3 种黄酮醇的细胞保护能力大小顺序为槲皮素>杨梅黄酮>山奈素。结果显示,黄酮醇的细胞保护能力大小与其分子结构中B 环上羟基数目的多少无关。     

桑叶生物碱粗提物对D-半乳糖诱导的小鼠蛋白氧化损伤的改善作用

目的:探讨桑叶生物碱改善小鼠蛋白氧化损伤作用效果与机理,旨在为桑叶生物碱的开发利用提供理论依据。方法:采用D-半乳糖(D-galactose,D-Gal)诱导建立小鼠氧化损伤模型,灌胃不同剂量的桑叶生物碱,于第8周末测定小鼠血浆蛋白羰基(protein carbonyl,PCO)、晚期蛋白氧化产物(advanced oxidation protein products,AOPP)、3-硝基酪氨酸(3-nitrotyrosine,3-NT)含量及超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)、醌氧化还原酶1(NAD(P)H quinine oxidoreductase 1,NQO1)活力,实时荧光定量聚合酶链式反应测定肝脏组织SOD、GSH-Px、NQO1、核因子E2相关因子2(nuclear erythroid related factor 2,Nrf2)、Kelch样环氧氯丙烷相关蛋白1(Kelch like ECH-associated protein-1,Keap1)mRNA表达...     

Quercetin attenuates ethanol-derived microsomal oxidative stress: Implication of haem oxygenase-1 induction

As the main source of reactive oxygen species (ROS), hepatic microsomes are susceptible to ROS attack, especially upon CYP2E1 activation by ethanol. The objective of this study was to evaluate the hepatoprotective effect of quercetin, by inducing haem oxygenase-1 (HO-1), on ethanol-induced microsomal oxidative stress. Chronic alcohol administration to adult rats (4.0 g/kg for 90 days) resulted in microsomal redox disturbance and liver dysfunction, accompanying CYP2E1 upregulation and HO-1 downregulation of both protein expression and enzymatic activity. Quercetin (100 mg/kg) induced HO-1, which was not completely suppressed by ethanol. Moreover, quercetin pretreatment to ethanol-fed rats lowered CYP2E1 induction, partially normalised ethanol-overwhelmed microsomal antioxidative system, decreased ROS level and lipid peroxidation, and alleviated the leakage of transaminases. Given the beneficial effect of HO-1, its induction by quercetin may contribute to the protective role against CYP2E1-mediated oxidative stress on hepatic microsomes.     

A single nucleotide polymorphism in the CYP2E1 gene promoter affects skatole content in backfat of boars of two commercial Duroc-sired crossbred populations

The prevention of unpleasant boar taint is the main reason for castration of male piglets. This study aimed to investigate how the malodorous compound skatole is affected by a single nucleotide polymorphism (g.2412 C>T at -586 ATG) in the porcine cytochrome p450 II E1 (CYP2E1) gene. 119 boars of two commercial Duroc-sired crossbred populations raised at different farms were investigated. Skatole and androstenone in backfat averaged 114±125ng/g and 1206±895ng/g melted fat, respectively. The frequency of the genotypes CC, CT, and TT was 25, 52, and 23%, respectively. CC boars had the highest average skatole levels (175ng/g) compared to CT (92ng/g) and TT (93ng/g). Applying suggested sensory threshold levels for skatole (>150ng/g) and androstenone (>2000ng/g), 30% of the carcasses may be unacceptably tainted while the proportion of tainted carcasses is significantly higher within genotype CC (56.7%) compared to genotypes CT (24.3%) and TT (14.8%). Effective reduction of tainted carcasses appears feasible applying marker assisted selection.     

Protective effects of protein hydrolysate from marine microalgae Navicula incerta on ethanol-induced toxicity in HepG2/CYP2E1 cells

Ethanol is independently known to cause tissue damage through various mechanisms. This study was designed to evaluate the protective effect of marine microalgae, Navicula incerta protein enzymatic hydrolysates (NEHs) against ethanol-induced hepatotoxicity in HepG2 cells transfected with human CYP2E1. Induction of CYP2E1 by ethanol is one of the central pathways by which ethanol generates a state of oxidative stress in hepatocytes. When the alcohol-induced cells were treated with NEHs at various concentrations, there was a dose-dependent decrease of gamma-glutamyl transpeptidase (GGT) activity in the culture media and loss of cell viability. Among the NEHs constituents the hydrolysates which were obtained by papain (P-NEH), pronase-E (PR-NEH) and α-chymotrypsin (A-NEH) activity attenuated the ethanol cytotoxicity effectively, respectively. The activity appeared to be a GGT inhibitor. Therefore, the cytoprotective effects at alcohol-induced HepG2/CYP2E1 cells could be attributed to the inhibition of GGT activity by NEHs. This study suggests that NEHs have enough potential to be considered as highly active compounds against ethanol toxicity which leads NEHs to be significant nutraceuticals.     

Effects of sex, weight, diet and hCG administration on levels of skatole and indole in the liver and hepatic activities of cytochromes P4502E1 and P4502A6 in pigs

Cytochromes P4502E1 (CYP2E1) and P4502A6 (CYP2A6) catalyse metabolic reactions of skatole and indole metabolism. The objectives of this study were as follows: to evaluate whether activities of CYP2E1 and CYP2A6 in pigs of two live weights (LW) differ between males and females; to investigate whether activities of CYP2E1 and CYP2A6 are affected by hCG stimulation; and to investigate whether the levels of skatole and indole in the liver and the activities of CYP2E1 and CYP2A6 are affected by raw potato starch (RPS). Female pigs expressed higher CYP2A6 activity at 90kg LW, and higher CYP2E1 activity at 115kg LW compared to male pigs. Skatole levels in the liver were higher in male pigs than in female pigs at both LW, whereas indole levels were higher in males only at 115 kg LW. Neither levels of indolic compounds in the liver nor enzyme activities were affected by hCG stimulation. The inclusion of RPS in the diet reduced skatole levels in the liver in both sexes and increased CYP2A6 activity in female pigs. It was concluded that the incidence of boar taint may depend on both skatole amount, which reach the liver, and the activities of enzymes involved in skatole metabolism, which may vary depending on sex, live weight, and diet.     

The effects of T-2 toxin exposure on liver drug metabolizing enzymes in rabbit

High doses of T-2 toxin are known to decrease protein synthesis and mono-oxygenase activities in rat liver. The purpose of this study was to investigate whether exposure at a low dose could alter the normal metabolism of the xenobiotic by the liver. Three doses of T-2 toxin, dissolved in olive oil, were orally and daily administered to New Zealand white rabbits for five days. At 0.5mg/kg, three of the five animals died, whereas only a weak decrease in body weight gain and moderate signs of toxicity occurred in rabbits receiving 0.25mg/kg/day, and the body weight increased without signs of toxicity at 0.1mg/kg/day. At 0.25mg/kg/day, total liver microsomal P450 content, and the activities of aminopyrine and benzphetamine N-demethylases, pentoxyresorufin O-depentylase, glutathione S-transferases accepting 1-chloro-2,4-dinitrobenzene and 1,2-dichloro-4-nitrobenzene as substrates, were decreased. By contrast, ethylmorphine and erythromycin N-demethylases, ethoxyresorufin and methoxyresorufin O-dealkylases, aniline hydroxylase, and UDP-glucuronyltransferase accepting p-nitrophenol as substrate, were unaffected. The expression of P450 1A1, 1A2, 2A1, and 2B4, but not P450 2C3 and 3A6, were also decreased, whereas microsomal conjugated dienes, fluorescent substances, and malondialdehyde contents were increased. At 0.1mg/kg/day, neither significant effects on drug metabolizing enzymes nor microsomal oxidative damages were obtained. Taken together, these results suggest that a short exposure time to the mycotoxin would not be associated with significant changes in the normal metabolism of xenobiotics by the liver.     

The relationship between oxidative damage and vitamin E concentration in blood, milk, and liver tissue from vitamin E supplemented and nonsupplemented periparturient heifers

This study investigated the relationship between oxidative damage and the effect of vitamin E supplementation in blood, milk, and liver tissue in 16 periparturient heifers. The question is whether measurements of oxidative and vitamin E status in blood of a periparturient cow are representative of the total body, given that blood concentrations of both vitamin E and oxidative stress products change around this period. The daily vitamin E intake of the vitamin E-supplemented Holstein-Friesian heifers (n = 8) was 3,000 international units and was started 2 mo before calving; the control heifers (n = 8) were not supplemented. Oxidative damage was determined on the basis of malondialdehyde (MDA) concentrations. Blood was sampled 9 times before calving, on calving day, and twice after calving. Liver biopsies were taken at wk −5, −1, and 2 relative to calving day. Milk was obtained from all heifers immediately after calving, the first 2 milkings and on d 3, 7, and 14 at 0600 h. Serum and liver tissue were analyzed for vitamin E, cholesterol, and MDA; and milk samples were analyzed for vitamin E, MDA, fat, protein, and somatic cell count. The results showed that vitamin E supplements increased both absolute vitamin E concentrations and the ratio of vitamin E to cholesterol in blood and liver tissue. Absolute vitamin E concentration in milk tended to be greater in supplemented cows. Based on the increased MDA blood concentrations at calving, it seems that dairy heifers experience oxidative stress. The effect of vitamin E on MDA differs between the blood, liver, and mammary gland. Vitamin E supplementation could not prevent the increase in blood MDA at calving, but the significantly lower MDA blood concentrations of supplemented cows in the 2 wk after calving suggest that vitamin E has a role in recovery from parturition-related oxidative stress. Vitamin E supplementation reduced oxidative damage in liver, whereas no obvious effect was found on milk MDA concentrations. A strong relationship was found between blood and liver vitamin E and the ratio of vitamin E to cholesterol. Concentrations of MDA in blood and milk were also strongly related. The results show that the relationship between oxidative damage and vitamin E differs within blood, liver tissue, and milk. This implies that oxidative and vitamin E status calculated on the basis of blood values alone should be interpreted with caution and cannot be extrapolated to the whole animal.     

Effects of ready to drink teas on AhR- and PXR-mediated expression of cytochromes P450 CYP1A1 and CYP3A4 in human cancer cell lines and primary human hepatocytes

A variety of xenobiotics are taken in the diet and they can interfere with regulatory pathways of drug metabolizing enzymes in humans. This can result in food-drug interactions, which is undesirable clinical situation where drug pharmacokinetics are influenced by dietary compounds. Xenobiotics-mediated food-drug interactions include the induction of drug metabolizing cytochromes P450. The expression of the most important inducible cytochromes CYP1A and CYP3A4 are regulated by xenoreceptors PXR and AhR.We examined extracts from 17 different flavoured ready to drink teas (RDTs) for their capabilities to activate PXR and AhR receptors and to induce CYP3A4 and CYP1A genes. Primary cultures of human hepatocytes and cancer cell lines HepG2 and LS174T were used as in vitro models. Gene reporter assays, RT-PCR and Western blots were performed.We identified three RDTs that induced CYP3A4 mRNA and protein, implying a potential for food-drug interactions. Several RDTs slightly elevated CYP1A1 expression or activated AhR.     

Docosahexaenoic acid downregulates phenobarbital-induced cytochrome P450 2B1 gene expression in rat primary hepatocytes via the c-Jun NH2-terminal kinase mitogen-activated protein kinase pathway

Mitogen-activated protein kinase (MAPK) pathways play central roles in the transduction of extracellular stimuli into cells and the regulation of expression of numerous genes. Docosahexaenoic acid (DHA) was shown to be involved in the regulation of expression of drug metabolizing enzymes (DMEs) in rat primary hepatocytes in response to xenobiotics. Cytochrome P450 2B1 (CYP 2B1) is a DME that is dramatically induced by phenobarbital-type inducers. The constitutive androstane receptor (CAR) plays a critical role in regulating the expression of DMEs, and the phosphorylation/dephosphorylation of CAR is an important event in CYP 2B1 expression. In the present study, we determined the effect of DHA on MAPK transactivation and its role in CYP 2B1 expression induced by phenobarbital. c-Jun NH2-terminal kinase (JNK) JNK1/2 and ERK1/2 were activated by phenobarbital in a dose-dependent manner. DHA (100 muM) inhibited JNK1/2 and ERK2 activation induced by phenobarbital in a time-dependent manner. Both SP600125 (a JNK inhibitor) and SB203580 (a p38 MAPK inhibitor) inhibited CYP 2B1 protein and mRNA expression induced by phenobarbital. SB203580 significantly increased the intracellular 3'-5'-cyclic adenosine monophosphate (cAMP) concentration compared with a control group (p < 0.05). Our results suggest that inhibition of JNK activation by DHA is at least part of the mechanisms of DHA's downregulation of CYP 2B1 expression induced by phenobarbital.