补肾活血方对复发性流产小鼠蜕膜组织VEGF/PI3K/Akt通路表达的影响

Effect of Bushen Huoxue Formula on expression of VEGF/PI3K/Akt pathway in decidua of mice with recurrent abortion

目的观察补肾活血方对复发性流产小鼠蜕膜组织VEGFA、VEGFR2及对PI3k/Akt信号转导通路的影响,探讨其治疗复发性流产的可能作用机制。方法建立BALB/c×CBA/J正常妊娠小鼠模型及DBA/2×CBA/J复发性流产小鼠模型,实验随机分为正常组、模型组、黄体酮组(3.33 mg/kg)、补肾活血方低剂量组(10.47 g/kg)、补肾活血方中剂量组(20.93 g/kg)、补肾活血方高剂量组(41.87 g/kg),各组给予相应药物灌胃2周。观察各组不同浓度药物干预后CBA/J小鼠的胚胎丢失率;采用免疫组化检测小鼠蜕膜组织VEGFA、VEGFR2蛋白的表达;采用Western blot检测小鼠蜕膜组织VEGFA、VEGFR2、PI3K、Akt、p-Akt蛋白的表达。结果(1)与模型组比较,黄体酮组和补肾活血方低、中、高剂量组小鼠胚胎丢失率均明显下降(P<0.05)。(2)免疫组化结果表明,与模型组比较,黄体酮组和补肾活血方中、高剂量组VEGFA蛋白阳性表达增强,黄体酮组和补肾活血方低、中、高剂量组VEGFR2蛋白阳性表达增强(P<0.05);与补肾活血方低剂量组比较,补肾活血方中、高剂量组VEGFA蛋白阳性表达增强(P<0.05)。(3)Western blot结果表明,与模型组比较,黄体酮组和补肾活血方低、中、高剂量组VEGFA、VEGFR2、PI3K、p-Akt蛋白表达水平明显升高(P<0.05);与补肾活血方低、中剂量组比较,补肾活血方高剂量组VEGFA、VEGFR2、PI3K、p-Akt蛋白表达水平明显升高(P<0.05)。结论补肾活血方能上调复发性流产模型小鼠蜕膜组织VEGFA及其受体VEGFR2的表达,其机制可能与激活PI3K/Akt信号通路相关。

Objective To observe the effect of Bushen Huoxue Formula on VEGFA,VEGFR2 and PI3 K/Akt signal transduction pathway in decidua of recurrent abortion mice,and to explore the possible mechanism of Bushen Huoxue Formula in treating recurrent abortion.Methods BALB/c×CBA/J normal pregnant mice model and DBA/2×CBA/J recurrent abortion mice model were established.The mice were randomly divided into normal group,model group,progesterone group(3.33 mg/kg),and Bushen Huoxue Formula low-dose(10.47 g/kg),medium-dose(20.93 g/kg)and high-dose(41.87 g/kg)groups.Each group was given corresponding drugs by gavage for 2 weeks.The embryo loss rate of CBA/J mice after different concentrations of drug intervention was observed;the expressions of VEGFA and VEGFR2 proteins in decidua were detected by immunohistochemistry;the expressions of VEGFA,VEGFR2,PI3 K,Akt,p-Akt proteins in decidua were detected by Western blot.Results(1)Compared with the model group,the embryo loss rates of the progesterone group and the low-dose,medium-dose and high-dose groups of Bushen Huoxue Formula were significantly decreased(P<0.05).(2)The results of immunohistochemistry showed that:compared with the model group,the positive expression of VEGFA protein was increased in the progesterone group,and the medium-dose and high-dose groups of Bushen Huoxue Formula;the positive expression of VEGFR2 protein was increased in the progesterone group and the low-dose,medium-dose and high-dose groups of Bushen Huoxue Formula(P<0.05);compared with the low-dose group of Bushen Huoxue Formula,the positive expression of VEGFA protein in the medium-dose and high-dose groups of Bushen Huoxue Formula was increased(P<0.05).(3)Western blot results showed that:compared with the model group,the expressions of VEGFA,VEGFR2,PI3 K,p-Akt proteins in the progesterone group,and the low-dose,medium-dose and high-dose groups of Bushen Huoxue Formula were significantly increased(P<0.05);compared with low-dose and medium-dose groups of Bushen Huoxue Formula,the protein expressions of VEGFA,VEGFR2,PI3 K and pAkt in the high-dose group of Bushen Huoxue Formula were significantly increased(P<0.05).Conclusion Bushen Huoxue Formula can up-regulate the expression of VEGFA and VEGFR2 in decidua of recurrent abortion model mice,and the mechanism may be related to the activation of PI3 K/Akt signaling pathway.