摘要
目的:以雌激素效应相关因子、血管生成相关因子及其相互作用为靶点,探讨在子宫腺肌病(AM)小鼠疾病发生发展的不同时间段及子宫在位及异位内膜不同空间部位,蒲灵化瘀止痛方治疗AM的作用机制。方法:对8周龄ICR雌鼠施行雄鼠垂体宫腔内移植手术,术后分别饲养3个月及6个月,形成M1组及M2组(AM小鼠模型3月组及6月组)。选择蒲灵化瘀止痛方为实验药物,同时将未造模同期小鼠设为正常组。各组分别给药3个月(Z1组及Z2组)。运用免疫组化技术观察雌激素效应相关因子(P450、COX-2、ERa)及血管生成相关因子(VEGF、ENS、CD31、MMP-2)在模型组、用药组随疾病发生发展的不同时间段及子宫在位及异位内膜不同空间部位的变化。结果:在位内膜方面,1与正常组比较:M1及M2组CD31显著下降(P<0.01,P<0.05)、VEGF显著增高(P<0.001)、MMP-2显著增高(P<0.01);P450、COX-2、ERa表达在M1组无显著差异,P450、COX-2在M2组显著增高(P<0.001)。2M1与M2组比较:P450、COX-2、ERa表达M2组显著高于M1组(P<0.01,P<0.001,P<0.05)。3治疗前后比较:Z1与M1比较,VEGF、MMP-2表达显著下降(P<0.01,P<0.05),CD31显著升高(P<0.001);Z2与M2比较,P450、COX-2、ERa表达下降显著(P<0.01,P<0.001,P<0.05)。在异位内膜方面,1与正常组比较:M1及M2组P450、COX-2表达及M2组ERa值表达均显著增高(P<0.001,P<0.01);2M1与M2组比较:M2组COX-2、ERa、P450表达与M1组比较有增高趋势(P=0.001,P=0.051,P=0.122);3治疗前后比较:Z1与M1比较,P450显著下调(P<0.05)、COX-2及ERa无显著变化;Z2与M2比较,P450、COX-2、ERa下降显著(P<0.001,P<0.001,P<0.05)。结论:蒲灵化瘀止痛方可多靶点治疗AM。病程早期的中药治疗主要以血管生成相关因子为靶点,改善早期在位内膜的缺血缺氧状况,直接截断由此触发的雌激素生成的正反馈链。疾病的后阶段,中药治疗靶点主要是在位及异位内膜雌激素效应相关因子,通过雌激素效应的变化影响血管生成因子的变化。
Objective： Observe the express of estrogen effect correlator and angiogenesis correlator, investigate manage mechanism of Puling Huyu Zhitong Formula （PLF） in AM mice during the different period of the development of the disease and in the different position of the endomembrane.. Method： Build the AM mouse model by pituitary intrauterine transplant operation. Divide the mice into three groups on random： the normal group（not take the operation）, group M1 （three months after the operation）, group M2 （six months after the operation）.Select pulinghuayuzhitongfang （PLF） as the experimental drug, set up group Z1 and Z2 （which is administered drug to M1, M2 for 3 months separately）. Detect the express of estrogen effect correlator and angiogenesis correlator in eutopic and ectopic site of endometrium respectively by method of immunohistochemisty dyeing. Result： In the eutopic site of endometrium： （1） Compared to normal group, the express of CD31 decreased notably （P〈0.01,P〈0.05）, VEGF, MMP-2 increased significantly in both M1 and M2 group （P〈0.001, P〈0.01,）. There was no notable change of P450, ERa, COX-2 in group M1 contrast to normal group, but there was a dramatic up-regulation of P450, COX-2 in group M2 contrast to normal group （P〈0.001）. （2） The express of P450, COX-2 and ERa was more higher in group M2 than in group M1 （P〈0.01, P〈0.001, P〈0.05）. （3） After treatment, there was a significant down-regulate of VEGF, MMP-2 （P〈0.01, P〈0.05） and up-regulate of CD31 （P〈0.001） in group Z1 contrast to MI; the express of P450,COX-2,ERa had a dramatic descend in group Z2 contrast to group M2 （P〈0.01, P〈0.001, P〈0.05）, however, which was not happened in the group ZI contrast to M1, except in the express of P450. In ectopic site, （1） the express of P450, COX-2 in group M1 and M2, ERa in group M2 increased notably contrast to normal group （P〈0.001, P〈0.01）, （2） there was a increased trend in the express of COX-2, ERa, P450 in group M2 contrast to the M1 （P=0.001, P=0.051, P=0.122）. （3） After treatment, the level of P450, COX-2, ERa desended siginificantly in group Z2 contrast to Z2 （P〈0.001, P〈0.05）, meanwhile, the level of P450 noteblely changed in group Z1 contrast to M1 （P〈0.05）, while which was not happened in express of COX-2, ERa. Conclusion： PLF can treat the AM mice in multitarget point. In the early stage of the disease, the target of the treatment is the angiogenesis correlator, improve the situation of ischemia and anoxia in the eutopia site of the endometrium, stop the positive feedback loop of COX-2-PGE2-P450arom. In the later stage of the disease, the major target of the treatment of the drug is the estrogen effect factor, result in regulate the chang of the angiogenesis correlator.
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2015年第6期2164-2167,共4页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家自然科学基金项目(No.81273796)~~
