基于PI3K/AKT信号通路探讨琥珀散对子宫内膜异位症小鼠的影响及作用机制研究

To investigate the effect and mechanism of Amber powder on endometriosis mice based on PI3K/AKT signaling pathway

目的探讨琥珀散干预磷脂酰肌醇3-激酶/蛋白激酶B(phosphatidy linositol 3-kinase/protein kinase B,PI3K/AKT)信号通路对子宫内膜异位症发生发展的影响及机制。方法选用BALB/c小鼠,随机分为正常组、疾病模型组、琥珀散组、西药组,每组10只。造模后琥珀散组按成人临床用药量等效量的2倍给予14.3 g/kg灌胃,连续给药4周;西药组将LY294002溶于二甲基亚砜后腹腔注射,以0.04 g/kg进行计算,每周腹腔注射一次,给药4周;正常组、疾病模型组给予相同体积的生理盐水灌胃,连续给药4周。给药结束后,采用ELISA法检测各组小鼠血清内TP53、丝氨酸苏氨酸蛋白激酶B1(serine/threonine protein kinase 1,AKT1)、血管内皮生长因子A(vascular endothelial growth factor-A,VEGF-A)和丝裂原激活蛋白激酶3(mitogen-activated protein kinase 3,MAPK3)的含量,采用免疫组化法检测各组小鼠内膜组织中PI3K、AKT的表达。结果与正常组相比,疾病模型组血清TP53、AKT1、VEGF-A和MAPK3的含量均显著升高(P<0.01),琥珀散组与西药组血清TP53、AKT1、VEGF-A和MAPK3的含量无明显差异(P>0.05);与疾病模型组相比,琥珀散组与西药组血清TP53、AKT1、VEGF-A和MAPK3的含量显著降低(P<0.01)。与正常组相比,疾病模型组异位内膜组织PI3K、AKT的蛋白含量表达均显著升高(P<0.01);与疾病模型组相比,琥珀散组、西药组异位内膜组织PI3K、AKT的蛋白含量表达显著降低(P<0.01)。结论琥珀散可下调TP53、AKT1、VEGFA和MAPK3的表达,抑制PI3K/AKT信号通路,而发挥对EMs的多靶点调控作用。

Objective To explore the mechanism of Amber powder in intervening PI3K/AKT signaling pathway on the occurrence and development of endometriosis.Methods BALB/c mice were selected and randomly divided into normal group,disease model group,Amber powder group and western medicine group,with ten rats in each group.Except the normal group,the other groups were modeled by intraperitoneal injection.Endometrial fragments of donated mice were injected into the abdominal cavity of mice to establish the endometriosis model mice.After buliding,the Amber powder group was given 14.3 g/kg by gavage at twice the equivalent dose of adult clinical dosage for 4 weeks;the western medicine group was given LY294002 dissolved in DMSO and injected intraperitoneally at 0.04 g/kg,once a week,for 4 weeks;rats in the normal group and the model group were given the same volume of normal saline by gavage for 4 weeks.After administration,the serum levels of TP53,AKT1,VEGFA and MAPK3 were detected by ELISA,and the expressions of PI3K and AKT in the intimal tissue of each group were detected by immunohistochemistry.Results The serum levels of TP53,AKT1,VEGFA and MAPK3 in the disease model group were significantly increased,compared with the normal group(P<0.01),while there was no significant difference in the serum levels of TP53,AKT1,VEGFA and MAPK3 between the Amber powder group and the western medicine group(P<0.01).Comparison with the disease model group,the levels of TP53,AKT1,VEGFA and MAPK3 in the serum of Amber powder group and the western medicine group were significantly decreased(P<0.01).The protein expressions of PI3K and AKT in the ectopic endometrial tissue of the disease model group were increased conspicuously than the normal group(P<0.01).AKT protein expression was significantly decreased(P<0.01).Conclusion Amber powder could down-regulate the expressions of TP53,AKT1,VEGFA and MAPK3,inhibit the PI3K/AKT signaling pathway,and play a multi-target regulatory effect on EMs.