安徽医科大学硕士学位论文非诺贝特、吡格列酮对 SD 大鼠 Visfatin 蛋白表达的影响中文摘要目的研究非诺贝特、吡格列酮对 SD 大鼠 Visfatin 蛋白表达的影响。方法 120只 6 周龄雄性 SD 大鼠随机分为普食组和高脂组，每组 60 只。普食组给予普通饲料喂养，高脂组予高脂饲料。定期称重大鼠，12 周肥胖大鼠形成模型后，药物干预组(普食+非诺贝特组、高脂+非诺贝特组、普食+吡格列酮组、高脂+吡格列酮组,每组 20 只）分别予非诺贝特 30mg/kg.d、吡格列酮 10mg/kg.d 灌胃 4 周，余对照组(普食组、高脂组，每组 20 只）予相应溶剂灌胃，容积分别为 2ml/kg.d。16 周时抽血备测血清 Visfatin、游离脂肪酸(FFA）、甘油三酯(TG)、胆固醇(TCH）、空腹血糖(FBG)、空腹胰岛素(FINS）等指标，分离附睾周围脂肪、肾周脂肪、网膜脂肪(三者合称为内脏脂肪组织）和腹部皮下脂肪，称重并提取蛋白质，用 Westernblot 法检测 Visfatin 蛋白表达；分离腓肠肌组织，用 Western blot 法检测Visfatin 蛋白量和免疫组化方法检测肌肉组织中 Visfatin 蛋白的定位。结果 (1)与普食组相比：高脂组大鼠体重(BW)、内脏脂肪重量、皮下脂肪重量、血清Visfatin、FBG、FINS、TG、FFA、HOMA-index 均明显增高 (P0.05)； TCH 差异无统计学意义；普食组、普食+非诺贝特组和普食+吡格列酮组大鼠 BW、内脏脂肪重量、皮下脂肪重量、血清 Visfatin、FBG、FINS、TG、FFA、TCH、HOMA-index差异无统计学意义。与高脂组相比：高脂+非诺贝特组 BW、内脏脂肪重量、血清Visfatin、FBG、FINS、TG、FFA、HOMA-index 均显著下降 (P0.05)，皮下脂肪重量、TCH 也下降，但差异无统计学意义；高脂+吡格列酮组较高脂组血清 Visfatin、FBG、FINS、TG、FFA、HOMA-index 均显著下降 (P0.05)，BW 和皮下脂肪重量升高 (P0.05),内脏脂肪重量下降，但差异无统计学意义；(2)用 Western blot 法在内脏脂肪、皮下脂肪和肌肉组织中都检测到 Visfatin 蛋白的表达，各组动物内3安徽医科大学硕士学位论文脏脂肪组织中 Visfatin 表达均高于皮下脂肪组织 (P0.05)，皮下脂肪组织中Visfatin 表达高于肌肉组织 (P0.05)；与普食组相比，普食+非诺贝特组、普食+吡咯列酮组的内脏脂肪、皮下脂肪、肌肉组织中 Visfatin 表达差异无统计学意义。与高脂组相比，高脂+非诺贝特组、高脂+吡咯列酮组内脏脂肪 Visfatin 表达下降(P0.05),但皮下脂肪组织、肌肉脂肪组织中 Visfatin 表达差异无统计学意义。(3)免疫组化显示：肌肉组织中 Visfatin 主要表达于肌肉细胞而非肌肉间质的脂肪细胞或其他成纤维细胞等。(4)相关分析显示：血清 Visfatin 与内脏脂肪重量、内脏脂肪中 Visfatin 表达、FBG、HOMA-index、TG 呈正相关。结论 (1)高脂饮食可诱导大鼠产生营养性肥胖，出现糖脂代谢紊乱；非诺贝特和吡格列酮可以调节高脂饮食诱导的大鼠血脂代谢紊乱、改善胰岛素抵抗，同时非诺贝特可以降低肥胖大鼠体重；该两种药物对正常饮食状态下大鼠血糖、血脂无调节作用。(2)Visfatin 广泛表达于内脏脂肪、皮下脂肪和肌肉组织，其是主要表达于内脏脂肪的脂肪因子；(3)在高脂饮食诱导 SD 大鼠出现肥胖、胰岛素抵抗状态时，血清和内脏脂肪 Visfatin 出现代偿性高表达，非诺贝特和吡格列酮在改善糖脂代谢紊乱同时伴随有血清和内脏脂肪 Visfatin 表达下降。非诺贝特和吡格列酮对普通饮食状态下 Visfatin 表达无明显调节作用。关键词Visfatin 胰岛素抵抗 非诺贝特 吡格列酮4安徽医科大学硕士学位论文Effects of fenofibrate、pioglitazone on Visfatin protein expressionin SD ratsAbstractObjective: To investigate the effects of fenofibrate、pioglitazone onVisfatin proteinexpression in SD rats.Methods: 120 6-wk-old male SD rats were divided into 2 groups randomly : NC groupand HF group (n=60). NC group were fed with normal chow and the others withhigh-fat diets. After 12 weeks, models of obese rats were established, then the drugtreated groups (NC + fenofibrate group, NC + pioglitazone group , HF + fenofibrategroup and HF + pioglitazone group, n=20) were treated with 30mg/kg fenofibrate,10mg/kg pioglitazone intragastric daily for 4 weeks, and the control groups withhomologous solvent. The volume of drug or solvent was 2ml/kg daily. Body weight wasmeasured regularly.Serum Visfatin 、free fatty acids (FFA)、triglyceride (TG)、totalcholesterol (TCH)、fasting blood glucose (FBG) and fasting insulin (FINS) weredetected after 16 weeks. Epididymis fat、kidney fat、omenta fat ( the three as visceral fat)and abdominal subcutaneous fat were weighed and harvested for protein extractionand western blot analysis. Gastrocnemius was also harvested for western blot andimmunohistochemistry.Results(1) Compared with NC group , body weight (BW)、visceral fat 、subcutaneousfat 、serum Visfatin、FBG、FINS、TG、FFA and HOMA-index of HF group wereelevated significantly (P0.05); BW、visceral fat、subcutaneousfat、serum Visfatin、FBG、FINS、TG、FFA、TCH and HOMA-index5were not affected安徽医科大学硕士学位论文by fenofibrate or pioglitazone in the rats fed normal chow (P0.05)；Compared with HFgroup, BW、visceral fat、serum Visfatin、FBG、FINS、TG、FFA and HOMA-index wereall downregulated in HF+fenofibrate group（P0.05)；Serum Visfatin、FBG、FINS、TG、FFA and HOMA-index were decreasedby pioglitazone in HF +pioglitazone group (P0.05)，and pioglitazone increased BWand subcutaneous fat (P0.05). Fenofibrate and pioglitazone could downregulatevisceral fat Visfatin in high-fat diets fed rats (P0.05). Both of the two drugs had no effect on Visfatin expression in thetissues of the animals fed normal chow. (3) Immunohistochemistry: in the muscle,Visfatin was expression in the muscle cells but not the interstitial tissue. (4) Correlationanalysis: serum Visfatin was positively correlated with the weight of visceral fat、Visfatin expression in visceral fat、FBG、HOMA-ind