卡托普利对大鼠非酒精性脂肪肝的作用及机制研究

发布时间：2018-04-10 20:39

本文选题：非酒精性脂肪肝病 + 卡托普利　；参考：《泸州医学院》2014年硕士论文

【摘要】：目的：观察卡托普利对非酒精性脂肪肝大鼠的治疗效果并探讨其作用机制。方法：将80只雄性SD大鼠按1：5比例随机分为2组：正常对照组与高脂组。除第1周外,整个喂养期间,正常对照组大鼠给予普通饲料,其余大鼠给予高脂饮食(普通饲料+10%猪油+2%胆固醇+0.2%胆酸钠),所有大鼠自由饮水和进食。造模6周后,将高脂组随机分为5组：模型对照组、卡托普利高、中、低剂量组和罗格列酮阳性对照组。分别给予相应大鼠等容量罗格列酮和高、中、低剂量卡托普利,给予正常对照组和模型对照组等容量蒸馏水,每天灌胃一次。给药6周后,所有大鼠禁食12小时,称重,用1%戊巴比妥钠(0.4 ml·100g-1)进行麻醉后处死。腹主动脉取血,用全自动生化分析仪检测其血清空腹血糖(Fasting blood glucose, FBG)、丙氨酸氨基转移酶(Alanine aminotransferase、天门冬氨酸氨基转移酶(Aspartate aminotransferase, AST)、总胆固醇(Total cholesterol, TC)、甘油三酯(Triglyceride, TG)水平；放射免疫分析法(Radioimmunoassay, RIA)检测血清空腹胰岛素(fasting insulin, INS)水平；硫代巴比妥酸(Thibabituric acid, TBA)法检测血清丙二醛(Malondialdehyde, MDA)水平、比色法检测血清谷胱甘肽(Glutathione, GSH)水平；酶联免疫吸附测定法(Enzyme-linked immunosorbent assay, ELISA)检测血清瘦素(Leptin, LEP)水平。分离肝脏并称量肝湿重,然后取相同部位肝组织作HE染色,于光镜下观察其病理学变化；取另一部分肝脏进行组织匀浆,采用酶偶联比色法检测其TC、TG含量。采用逆转录-聚合酶链反应(Reverse transcription-polymerase chain reaction, RT-PCR)技术检测肝组织中CYP2E1的mRNA表达水平。按公式计算胰岛素抵抗指数(Insulin resistance index, IRI)、胰岛素敏感指数(Insulin sensitive index, ISI)和肝指数(Liver index)。结果：模型对照组FBG、INS、TC、TG、LEP、ALT、AST、MDA含量、肝指数、IRI、肝组织CYP2E1的mRNA表达水平均增高,GSH含量和ISI下降,与正常对照组比较,有显著性差异(P0.01或P0.05)。卡托普利高剂量组FBG、INS、TC、TG、LEP、ALT、AST、MDA含量、肝指数、IRI及肝组织CYP2E1的mRNA表达水平均降低,GSH含量和ISI上升,与模型对照组比较,有显著性差异(P0.01或P0.05)；阳性对照组FBG、INS、TC、TG、LEP、MDA含量、肝指数、IRI及肝组织CYP2E1的mRNA表达水平均降低,ISI上升,与模型对照组比较,有显著性差异(P0.01或P0.05)。肝组织光学显微镜下病理学观察发现：模型对照组可见弥漫性脂肪变性,主要为大泡性脂肪变性,肝细胞水肿、细胞排列紊乱,并可见大量炎细胞浸润。卡托普利高剂量组可见少量脂肪变性,主要为小泡性脂肪变性。大多数肝细胞排列正常,部分细胞水肿,并可见少量炎细胞浸润。阳性对照组病理变化情况与卡托普利高剂量组相似,只是脂肪变性较多且有部分为大泡性脂肪变性。结论：非酒精性脂肪肝大鼠造模成功。卡托普利能减轻肝细胞脂肪变性,对非酒精性脂肪肝有一定的防治作用。卡托普利可通过改善机体胰岛素抵抗、增强其抗氧化应激能力而达到防治效果。
[Abstract]:Objective: To observe the therapeutic effect of Kato Pury on nonalcoholic fatty liver in rats and explore its mechanism. Methods: 80 male SD rats by 1:5 were randomly divided into 2 groups: normal control group and hyperlipidemia group. In first weeks, the whole feeding period, the rats in the normal control group received general feed, the remaining rats were given high fat diet (normal diet +10% lard +2% cholesterol +0.2% sodium cholate), free drinking and eating all the rats. After 6 weeks, high fat diet group were divided into 5 groups: model control group, Kato Pury high, low dose group and rosiglitazone positive control group respectively. Give the corresponding rat capacity of rosiglitazone and high, low dose of Kato Pury, given the normal control group and model control group were treated with equal volume of distilled water by gavage once a day. After 6 weeks, all rats were fasted for 12 hours, weighing, with 1% pentobarbital sodium (0.4 ml 100g-1) of Ma Drunk. After killing the blood from the abdominal aorta, the serum fasting blood glucose was detected by automatic biochemical analyzer (Fasting blood, glucose, FBG), Alanine aminotransferase (alanine aminotransferase, aspartate aminotransferase (Aspartate, aminotransferase, AST), total cholesterol (Total, cholesterol, TC), triglycerides (Triglyceride, TG) level; radioimmunoassay (Radioimmunoassay, RIA) detection of serum fasting insulin (fasting, insulin, INS) level of thiobarbituric acid (Thibabituric; acid, TBA) were used to detect the serum malondialdehyde (Malondialdehyde, MDA) level, serum glutathione ratio method (Glutathione, GSH); enzyme linked immunosorbent assay (Enzyme-linked immunosorbent assay, ELISA) detection of serum leptin (Leptin, LEP). The level of separating the liver and liver wet weight weighing, and then take the liver tissue HE staining, light To observe the pathological changes under microscope; another part of the liver tissue homogenate, colorimetric method was used to detect the TC enzyme coupling TG content. Using reverse transcriptase polymerase chain reaction (Reverse transcription-polymerase chain reaction, RT-PCR) the expression level of CYP2E1 in liver tissue were detected in mRNA. According to the formula for the calculation of insulin resistance index (Insulin resistance index, IRI), insulin sensitivity index (Insulin sensitive, index, ISI) and liver index (Liver index). Results: the model control group FBG, INS, TC, TG, LEP, ALT, AST, MDA content, liver index, IRI, the expression level of CYP2E1 in liver tissue of mRNA were increased, the content of GSH and ISI decreased, compared with normal control group, there was significant difference (P0.01 or P0.05). Kato Pury Gogh FBG INS, TC group, TG, LEP, ALT, AST, MDA content, liver index, the expression level of IRI and CYP2E1 in liver tissues of mRNA were decreased and the content of GSH and ISI increased, and The model group, there was significant difference (P0.01 or P0.05); positive control group FBG, INS, TC, TG, LEP, MDA content, liver index, the expression level of IRI and CYP2E1 in liver tissues of mRNA were decreased, ISI increased, compared with the model group, there was significant difference (P0.01 or P0.05) observation of optical microscope. Liver tissue pathological model control group showed diffuse fatty degeneration, mainly for bullous steatosis, liver cell edema, cell disorder, and the number of inflammatory cell infiltration. Kato Pury Gogh dose group showed a small amount of fat, mainly for microvesicular steatosis. Most liver cells were normal. Some cells edema, and a small amount of inflammatory cell infiltration. Positive control group pathological changes and Kato Pury Gogh dose were similar, only more fatty degeneration and partial bullous steatosis. Conclusion: nonalcoholic fatty liver disease in rats Kato Pury can reduce the steatosis of liver cells and have some preventive and therapeutic effects on non-alcoholic fatty liver disease. Kato Pury can improve the body's insulin resistance and enhance its ability of anti oxidative stress, so as to achieve the control effect.

【学位授予单位】：泸州医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R965

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