二甲双胍对多囊卵巢综合征大鼠排卵及卵巢颗粒细胞中B淋巴细胞瘤-2和兔抗人单克隆抗体表达的影响

Effect of metformin on ovulation of polycystic ovary syndrome rats and expressions of Bcl-2 and Bax in ovarian granule cells

目的探究二甲双胍(MET)对多囊卵巢综合征(PCOS)大鼠排卵及卵巢颗粒细胞中B淋巴细胞瘤-2(Bcl-2)和兔抗人单克隆抗体(Bax)表达的影响。方法用来曲唑(LTZ)皮下埋植方法诱导PCOS。称量大鼠体质量和卵巢质量,HE染色卵巢并在显微镜下进行形态学观察。采用放射免疫法(RIA)方法检测血清中促黄体生成素(LH)、雌二醇(E_2)、孕酮(P)和睾酮(T)含量水平。最后用实时聚合酶链反应(Real-time PCR)和蛋白质印迹法(western blotting)分别检测颗粒细胞中Bcl-2和Bax基因、蛋白的表达。结果 PCOS SD雌性大鼠出现体质量增长减慢、卵巢组织质量增加、血清中LH和T含量水平增加、E_2和P含量水平下降,差异有统计学意义(F=71.457、54.052、24.341、162.981、62.593、172.848,均P<0.05),具有PCOS典型症状。然而PCOS+MET组大鼠的上述症状均较PCOS组大鼠明显缓解。PCOS大鼠卵巢颗粒细胞中Bcl-2的表达下降,Bax的表达上升,差异有统计学意义(F=15.179、13.375,均P<0.05);MET上调PCOS大鼠卵巢颗粒细胞中Bcl-2的表达,下调Bax的表达。结论 MET通过增加PCOS大鼠卵巢颗粒细胞中Bcl-2的表达和降低Bax的表达,抑制颗粒细胞的凋亡,从而促进PCOS大鼠的排卵作用。

Objective To explore the effect of metformin on ovulation of polycystic ovary syndrome （PCOS） rats and expressions of Bcl-2 and Bax in ovarian granule cells.Methods PCOS was induced by subcutaneous implantation of letrozole.Body weight and ovary weight were weighed.HE staining of ovary was performed,ovarian morphology was observed under microscope.Radioimmunoassay （RIA） was used to detect the levels of serum luteinizing hormone （LH）,estradiol （E2）,progesterone （P）,and testosterone （T）.Real-time PCR and Western blotting were used to detect the expressions of Bcl-2 and Bax genes and proteins in ovarian granule cells,respectively.Results Female SD rats with PCOS has typical symptoms of PCOS,the increase of body weight slowed down,the weight of ovarian tissues increased,the levels of serum LH and T increased,the levels of E2 and P decreased,there were statistically significant differences （F=71.457,54.052,24.341,162.981,62.593,172.848,all P〈0.05）.The above-mentioned symptoms in PCOS rats treated by metformin were relieved significantly compared with PCOS rats.The expression of Bcl-2 in ovarian granule cells of PCOS rats decreased,while the expression of Bax in ovarian granule cells of PCOS rats increased,there were statistically significant differences （F=15.179,13.375,P〈0.05）.Metformin upregulated the expression of Bcl-2 and downregulated the expression of Bax in ovarian granule cells of PCOS rats.Conclusion MET can inhibit the apoptosis of ovarian granular cells and promote ovulation of PCOS rats by increasing Bcl-2 expression and decreasing Bax expression.