中文摘要：

以人工诱导卵巢颗粒细胞胰岛素抵抗（Insulin Resistance,IR）细胞模型,观察细胞内关键信号分子的表达以及胰岛素增敏剂的体外作用。通过沃曼青霉素（Wortmannin,WT）特异性地抑制胰岛素传导途径中磷脂酰肌醇-3激酶（PI-3K）通路,作用于体外培养的猪卵巢颗粒细胞48h后,再加入曲格列酮、二甲双胍、小檗碱、隐丹参酮48h,观察对IR的改善情况。以葡萄糖氧化酶法检测细胞对葡萄糖的消耗量,以蛋白印记法（Western Blot）、逆转录-聚合酶链反应（RT-PCR）和实时荧光PCR法,检测葡萄糖转运蛋白4（Glut4）、丝裂原激活蛋白激酶（MAPK）、核转录因子过氧化物酶体增殖物激活受体γ（PPAR-γ）表达。结果表明：胰岛素糖代谢途径被WT阻滞,IR细胞对培养基中葡萄糖的摄取明显抑制。二甲双胍、小檗碱和隐丹参酮,都能使葡萄糖摄取增加,但稍逊于曲格列酮。IR细胞的Glut4的表达下降,MAPK和PPAR-γ表达升高;胰岛素增敏剂可以部分逆转IR细胞的上述信号蛋白的表达异常,以曲格列酮作用最为显著。总之,WT阻断PI-3K途径产生IR,胰岛素增敏剂能改善IR,隐丹参酮和小檗碱增敏的作用类似二甲双胍,而曲格列酮作用最为显著。

英文摘要：

This paper studies the biological effects of key molecules and insulin-sensitizing agents on Insulin Resistance （IR） of porcine granulosa cells. The model of IR for porcine granulosa cells induced by wortmannin, the PI-3K inhibitor, was established in vitro culture for 48 hours. Then porcine granulosa cells of IR were treated by troglitazone, metfomline, berberine and cryptotanshinone, respectively, in the next 48 hours. The effects of insulin-sensitizing agents on IR were observed. The amount of 3H glucose uptake was assessed by glucose oxidase method. The protein and mRNA expression of Glut4, MAPK and PPAR-γ were assessed by Western Blot, RT- PCR and fluorescence RT-PCR. The results indicate that the IR of porcine granulosa cells is induced by wortmannin during insulin signaling. The glucose intake is significantly decreased with the treatment of wortmannin and is increased with the treatment of insulin-sensitizing agents. The effects of berberine, cryptotanshinone and metformine on decrease levels of glucose are not as significant as those of troglitazone. The protein and mRNA expression of Glut4, measured in porcine granulosa cells of IR, are down--regulated by immunofluorescence and RT-PCR, respectively and are upregulated by the treatment of insulin-sensitizing agents, as compared with control granulosa cells. The mRNA expression of MAPK and PPAR-γ are up-regulated by IR and down-regulated by the treatment of insulin-sensitizing agents as compared with the controls. It is concluded that Wortmannin can induce the IR of porcine granulosa cells. The effects of berberine and cryptotanshinone on decrease levels of glucose are similar to the effects of metformine.