利拉鲁肽对非酒精性脂肪肝大鼠模型疗效评价和作用机制研究

发布时间：2018-01-01 03:12

本文关键词：利拉鲁肽对非酒精性脂肪肝大鼠模型疗效评价和作用机制研究　出处：《安徽医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的(1)采用氢质子磁共振波谱(~1H-MRS)、病理及血清指标来评价利拉鲁肽治疗非酒精性脂肪肝大鼠模型的疗效;(2)探讨分析利拉鲁肽对非酒精性脂肪肝病模型大鼠血清及肝组织FGF21水平的影响以及可能机制。方法(1)清洁级雄性SD大鼠32只,随机分为高脂组20只和正常对照组12只,高脂组造模成功后随机分为利拉鲁肽组和安慰剂组,继续予高脂饮食并分别予皮下注射利拉鲁肽或生理盐水,对照组予普通饲料喂养。(2)干预16周末,~1H-MRS检测各组大鼠肝脏相对脂肪含量,并行肝组织病理HE染色及生化、肝组织匀浆等指标检测;(3)酶联免疫吸附测定法(Elisa)测定三组大鼠血清FGF21浓度,蛋白免疫印迹法(Western blot)测定肝组织细胞裂解液中FGF21含量;(4)蛋白免疫印迹法对肝细胞内PPAR a及PPAR r蛋白表达情况进行相对定量研究;(5)Image J软件分析蛋白条带的灰度值,IPP图像软件分析免疫组化蛋白相对含量;采用SPSS 17.0统计软件行正态分布性检验,采用Sigma plot12.5作图,用“?x±s”来表示符合正态分布的所有计量资料;采用单因素方差分析来分析组间差异,其中采用SNK法进行两两比较;采用多组独立样本秩和检验对有序定性资料进行检验。结果(1)与安慰剂组相比,利拉鲁肽组大鼠体质量、肝指数、胰岛素抵抗指数(HOMA-IR)、血清甘油三酯(TG)、胆固醇(CHO)、低密度脂蛋白(LDL)、空腹胰岛素(FINS)、谷丙转氨酶(ALT)、肝匀浆TG及CHO均明显下降,差异有统计学意义(P0.05);三组空腹血糖(FBG)、高密度脂蛋白(HDL)无明显差异;(2)与安慰剂组相比,利拉鲁肽组大鼠肝组织病理及~1H-MRS结果示肝脏脂肪变性程度明显减轻甚至恢复正常,肝内相对脂肪含量明显减少(26.8±1.30 vs 13.0±0.71,P0.05);(3)血清中,安慰剂组FGF21浓度较利拉鲁肽组和正常对照组明显升高,(637.53±14.12 vs 509.49±18.84,P0.05),利拉鲁肽组与正常对照组差异无统计学意义(509.49±18.84 vs 503.98±20.23,P0.05);肝组织细胞裂解液中,安慰剂组FGF21含量明显低于利拉鲁肽组及正常对照组(1.38±0.23vs 2.50±0.78 vs 2.46±0.49,P0.05),利拉鲁肽组与正常对照组FGF21差异无统计学意义(2.50±0.78 vs 2.46±0.49,P0.05);(4)与安慰剂组相比,利拉鲁肽组大鼠肝组织PPARα表达明显增加(0.50±0.07 vs 0.67±0.06,P0.05),利拉鲁肽组和正常对照组差异无统计学意义(0.67±0.06 vs 0.71±0.07,P0.05);(5)与正常对照组相比,利拉鲁肽组和安慰剂组大鼠肝组织PPARγ表达明显减少(0.73±0.06 vs 0.65±0.56 vs 0.62±0.06,P0.05),利拉鲁肽组较安慰剂组表达较高,但差异无统计学意义(0.65±0.56 vs 0.62±0.06,P0.05)结论(1)利拉鲁肽能明显减轻高脂饮食诱导的NAFLD大鼠体质量,改善肝指数和肝脏相对脂肪含量;(2)利拉鲁肽改善NAFLD,促进FGF21的表达,其机制可能与利拉鲁肽上调肝组织PPARα表达有关;
[Abstract]:Objective (1) using proton magnetic resonance spectroscopy (~1H-MRS), and to evaluate the effect of serum index of pathological model of liraglutide in the treatment of nonalcoholic fatty liver rats; (2) investigate effects of liraglutide on serum FGF21 level and liver tissues of rats with non alcoholic fatty liver disease model and possible mechanisms. (1) male SD 32 rats were randomly divided into high-fat group 20 rats and 12 rats in normal control group, hyperlipidemia group rats were randomly divided into liraglutide group and the placebo group to a high-fat diet and then were treated with subcutaneous injection of liraglutide or saline, the control group was given common feed. (2) 16 weeks intervention ~1H-MRS detection of liver of rats relative fat content, hepatic tissue biochemical and pathological HE staining, detection of liver homogenate and other indicators; (3) enzyme-linked immunosorbent assay (Elisa) determination of serum FGF21 concentration of three protein group, India Trace method (Western blot) for determination of the content of FGF21 in liver tissue in cell lysate; (4) Western blot of liver cells in PPAR A and PPAR R protein expressions were used for relative quantitative study; (5) Image J software to analyze the gray level of protein bands, analysis of the relative content of protein by immunohistochemistry IPP image software; using SPSS 17 statistical software for test of normal distribution, using Sigma plot12.5 mapping, with "x + S?" said to comply with all normal distribution measurement data; using the single factor analysis of variance to analyze the differences between groups, of which 22 were compared by SNK method; using multiple sets of independent samples rank sum test. In order to test the qualitative data. Results (1) compared with the placebo group, body weight, liraglutide group rats liver index, insulin resistance index (HOMA-IR), serum triglyceride (TG), cholesterol (CHO), low density lipoprotein (LDL), fasting insulin (FINS), valley Alanine aminotransferase (ALT), liver homogenate TG and CHO were significantly decreased, the difference was statistically significant (P0.05); the three groups of fasting blood glucose (FBG), high density lipoprotein (HDL) had no significant difference; (2) compared with the placebo group, the liver tissue pathological group Liraru peptide and ~1H-MRS showed fatty liver degeneration was significantly reduced and even returned to normal, liver fat relative content decreased significantly (26.8 + 1.30 vs 13 + 0.71, P0.05); (3) serum FGF21 concentration was significantly higher than the placebo group, Liraru peptide group and normal control group, (637.53 + 14.12 vs 509.49 + 18.84, P0.05), peptide group and Liraru the control group had no significant difference (509.49 + 18.84 vs 503.98 + 20.23, P0.05); liver tissue in the cell lysate, the placebo group was obviously lower than that of Liraru FGF21 peptide group and normal control group (1.38 + 0.23vs 2.50 + 0.78 vs 2.46 + 0.49, P0.05), Liraru peptide group and normal control group FGF2 There was no significant difference between 1 (2.50 + 0.78 vs 2.46 + 0.49, P0.05); (4) compared with the placebo group, the expression of Liraru in liver tissue of rats PPAR alpha peptide significantly increased (0.50 + 0.07 vs 0.67 + 0.06, P0.05), Liraru peptide group and the control group had no significant difference (0.67 + 0.06 vs 0.71 + 0.07, P0.05); (5) compared with the normal control group, the expression of Liraru peptide group and the placebo group of rat liver tissue PPAR was decreased (0.73 + 0.06 vs 0.65 + 0.56 vs 0.62 + 0.06, P0.05), Liraru peptide group than in the placebo group was higher, but the difference was not statistically significant (0.65 0.62 + 0.06 + 0.56 vs, P0.05) conclusion (1) Liraru peptide can significantly reduce the body mass of NAFLD rats induced by high fat diet, improve the liver index and relative liver fat content; (2) Liraru peptides improve NAFLD, promote the expression of FGF21, and its mechanism may be related to liver tissues Liraru peptide PPAR expression;

【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R575;R-332

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