补肾活血方对复发性流产小鼠母胎界面血管微环境的影响

Effects of Bushen Huoxue Prescription on Vascular Microenviroment at Maternal-fetal Interface in Mice with Recurrent Spontaneous Abortion

目的观察补肾活血方对复发性流产(RSA)小鼠母胎界面血管微环境的影响,探讨其潜在机制。方法将CBA/J雌鼠与DBA/2J雄鼠2∶1合笼饲养建立RSA模型,同时将CBA/J雌鼠与BALB/c雄鼠合笼饲养建立正常妊娠作为正常组,将成模小鼠随机分为模型组、中药组和西药组,每组10只。妊娠第1日起,中药组予补肾活血方灌胃,西药组予地屈孕酮溶液灌胃,正常组及模型组灌胃等体积蒸馏水,连续13 d。计算各组小鼠胚胎丢失率,HE染色观察蜕膜及迷路组织病理变化,RT-PCR检测蜕膜组织透明质酸合成酶(HAS)mRNA表达,Westernblot检测蜕膜组织血管内皮生长因子(VEGF)、血管内皮钙黏蛋白(VE-cadherin)表达,硝酸还原酶法检测蜕膜组织一氧化氮(NO)含量,ELISA检测血清血栓素B2(TXB2)和6-酮前列腺素F1α(6-Keto-PGF1α)含量。结果与正常组比较,模型组小鼠胚胎丢失率升高(P<0.001),蜕膜细胞碎裂,血管数量减少,迷路滋养层细胞增生肥大,胞浆广泛空泡化,部分细胞变性、坏死;蜕膜组织HAS-1、HAS-2、HAS-3 mRNA表达和VEGF、VE-cadherin蛋白表达降低(P<0.05,P<0.01,P<0.001),NO含量减少(P<0.01),血清6-Keto-PGF1α含量减少(P<0.001)。与模型组比较,中药组小鼠胚胎丢失率降低(P<0.01),蜕膜细胞较完整,迷路滋养层细胞呈网状排列,母血窦较丰富,整体结构完整;蜕膜组织HAS-1、HAS-2、HAS-3 mRNA表达和VEGF、VE-cadherin蛋白表达升高(P<0.05,P<0.01),NO含量增加(P<0.01),血清6-Keto-PGF1α含量增加(P<0.001)。结论补肾活血方能通过增加促血管生成因子表达,降低血栓形成风险,改善小鼠母胎界面血管微环境,从而发挥对RSA的治疗作用。

Objective To observe the effects of Bushen Huoxue Prescription on vascular microenvironment at maternal-fetal interface in recurrent spontaneous abortion(RSA)model mice;To discuss its possible mechanism.Methods CBA/J female mice and DBA/2J male mice were reared in a 2:1 co-cage to establish the RSA model,while CBA/J female mice and BALB/c male mice were co-caged to establish a normal pregnancy as the normal group.The RSA model mice were randomly divided into model group,TCM group and Western medicine group,with 10 mice in each group.From the first day of pregnancy,the TCM group was given Bushen Huoxue Prescription by gavage,the Western medicine group was given didroxyprogesterone solution,and the normal group and model group were given the same volume of distilled water,for consecutive 13 days.The embryo loss rate of mice in each group was calculated,and the pathological changes of decidua and labyrinth tissues were observed by HE staining.The expression of HAS mRNA in decidual tissue was detected by RT-PCR.VEGF and VE-cadherin in decidual tissue were detected by Western blot.The content of NO in decidual tissue was detected by nitrate reductase method.The contents of TXB2 and 6-Keto-PGF1αin serum were detected by ELISA.Results Compared with the normal group,the embryo loss rate of mice in the model group was significantly increased(P<0.001).The decidual cells were broken and the number of blood vessels was reduced.The labyrinth trophoblast cells proliferated and hypertrophy,cytoplasm was extensively vacuolated,and some cells denaturated and necrosised.The expression of HAS-1,HAS-2,HAS-3 mRNA and VEGF,VE-cadherin protein in decidual tissue significantly decreased(P<0.05,P<0.01,P<0.001),the content of NO significantly reduced(P<0.01).The content of 6-Keto-PGF1αin serum significantly decreased(P<0.001).Compared with the model group,the embryo loss rate of mice in the TCM group was significantly decreased(P<0.01).The decidual cells were relatively complete,the labyrinth trophoblast cells were arranged in a reticular pattern,the maternal blood sinus was more abundant,and the overall structure was complete.The expression of HAS-1,HAS-2,HAS-3 mRNA and VEGF,VE-cadherin protein in decidual tissue significantly increased(P<0.05,P<0.01),the content of NO significantly increased(P<0.01).The content of 6-Keto-PGF1αin serum significantly increased(P<0.001).Conclusion Bushen Huoxue Prescription can play a therapeutic effect on RSA by increasing the expression of pro-angiogenic factors,reducing the risk of thrombosis,and improving the vascular microenvironment at the maternal-fetal interface in mice.