二甲双胍对复发性流产大鼠妊娠结局、内分泌及VEGF/MAPK信号通路的影响

Effects of metformin on pregnancy outcome,endocrine and VEGF/MAPK signaling pathway in rats with recurrent spontaneous abortion

目的探讨二甲双胍(Met)对复发性流产(RSA)大鼠血管内皮生长因子(VEGF)/丝裂原活化蛋白激酶(MAPK)信号通路及卵巢内分泌的影响。方法SD雌性大鼠与雄性大鼠按照2∶1的比例合笼交配后将受孕成功的雌性大鼠采用随机数字表法分为对照组,RSA组,Met低、中、高剂量组(250、1000、2000 mg/kg),黄体酮组(阳性对照组,156 mg/kg),每组10只。末次给药结束后24 h,将大鼠麻醉后处死,解剖大鼠子宫观察胚胎丢失情况;苏木精-伊红染色检测子宫蜕膜组织病理变化;放射免疫试剂盒检测血清激素水平;蛋白免疫印迹法检测子宫蜕膜组织中VEGF/MAPK通路蛋白水平。结果与对照组比较,RSA组大鼠死亡胚胎数及胚胎丢失率升高,血清卵泡刺激素(FSH)、黄体生成激素(LH)、垂体泌乳素(PRL)、雌二醇(E2)、孕酮(P)表达量以及子宫蜕膜组织中VEGF受体-2(VEGFR-2)、VEGF、磷酸化细胞外信号调节激酶(p-ERK)蛋白表达量均降低(P<0.05),子宫蜕膜组织受损严重;与RSA组比较,Met各剂量组大鼠死亡胚胎数及胚胎丢失率降低,血清FSH、LH、PRL、E2及P表达量以及子宫蜕膜组织中VEGFR-2、VEGF、p-ERK蛋白表达量均升高(P<0.05),子宫蜕膜组织损伤得到一定的缓解,且高剂量对上述指标的控制效果更好。结论Met可缓解RSA大鼠内分泌失调及子宫蜕膜损伤,提高VEGF/MAPK通路相关蛋白的表达,具有促进VEGF/MAPK通路激活的作用。

Objective To investigate effects of metformin(Met)on vascular endothelial growth factor(VEGF)/mitogenactivated protein kinase(MAPK)signaling pathway and ovarian endocrine in recurrent abortion(RSA)rats.Methods Female SD rats were mated with male rats in a ratio of 2∶1,then the female pregnant rats were randomly divided into the control group,the RSA group,the Met low-dose,middle-dose and high-dose groups(250,1000,2000 mg/kg)and the progesterone group(positive control group,156 mg/kg),with 10 rats in each group.At 24 hours after the last administration,rats were anesthetized and killed.The embryo loss was observed by dissecting rat uterus.Hematoxylin eosin(HE)staining was used to detect pathological changes of decidual tissue.Radioimmunoassay kit was used to detect serum hormone levels.The protein levels of VEGF/MAPK pathway in uterine decidua were detected by Western blot assay.Results Compared with the control group,the number of dead embryos and the rate of embryo loss were significantly increased in the RSA group,and serum levels of follicle-stimulating hormone(FSH),leuteinizing hormone(LH),pituitary prolactin(PRL),estradiol(E2),progesterone(P)and protein expressions of VEGF receptor-2(VEGFR-2),VEGF and phosphorylated extracellular signal-regulated kinase(p-ERK)in decidual tissue were significantly decreased(P<0.05),and the decidual tissue of uterus was seriously damaged.Compared with the RSA group,the number of dead embryos and the rate of embryo loss were significantly decreased in the Met groups.Serum levels of FSH,LH,PRL,E2,P and the protein expression levels of VEGFR-2,VEGF and p-ERK in decidual tissue were significantly increased(P<0.05).The damage of decidual tissue was alleviated,and high dose of Met showed better control effect on the above indexes.Conclusion Met can alleviate endocrine disorder and uterine decidua damage in RSA rats,which significantly improves the expression of VEGF/MAPK pathway related proteins,and promotes the activation of VEGF/MAPK pathway.