基于VDR及CYP27B1对冠脉通片拆方补肾活血作用靶点研究

Study on Guanmaitong and its components in invigorating the kidney and activating blood circulation action with VDR and CYP27B1

关键词:  冠脉通片  VDR  CPY27B1  补肾活血

Key words:  Guanmaitong tablet  VDR  CYP27B1  invigorating the kidney and activating blood circulation action

摘要:

[目的]探讨冠脉通片及其拆方基于维生素D受体（VDR）及1α-羟化酶（CYP27B1）对补肾活血的作用。[方法]采用"腺嘌呤+左冠状动脉前降支结扎法"制备肾阳虚心梗模型。将大鼠分为假手术对照组、肾阳虚心梗模型对照组、冠脉通片全方组、补肾拆方组、活血开窍拆方组、维生素D对照组。采用预防性给药，于AMI术前2 d灌胃相应受试物，术后给药5 d，每日1次。检测评价肾阳虚的血清肌酐（Scr）、三碘甲状腺原氨素（T3）、甲状腺素（T4）水平。酶联免疫法测血清活化1，25（OH）₂D₃水平。免疫组化法观察心室肌组织、肾组织VDR、肾组织CYP27B1表达。Western blot法检测心室肌组织、肾组织VDR和肾组织CYP27B1蛋白相对表达量。[结果]冠脉通片全方降低Scr作用明显（P<0.05），升高T3、T4作用明显（P<0.05）；可明显升高血清1，25（OH）₂D₃水平（P<0.05）。全方组肾组织CPY27B1蛋白表达增多，全方及补肾组可增加心肌和肾组织中的VDR表达。[结论]冠脉通片可通过CPY27B1和VDR作为核心环节，对其用于肾虚血瘀的补肾活血作用机制提供了基础研究的数据支持。

Abstract:

[Objective] To investigate the role of Guanmaitong tablets and its components in invigorating the kidney and activating blood circulation action with VDR and CPY27B1.[Methods] The model of myocardial infarction rats with kidney yang deficiency was established by adenine and ligation of the left anterior descends in branch.The rats were divided into sham operation group,myocardial infarction kidney yang deficiency model group,Guanmaitong tablets whole components group,invigorating the kidney components group, activating blood circulation components group and the vitamin D group.Precautionary administered orally prior to AMI patients 2 d,then administration 5 d,1 time of each day after model.Serum creatinine (Scr) and T3,T4 levels are detected.ELISA measurement of serum activation 1,25(OH)₂D₃ level.Immunohistochemical staining was observed in ventricular muscle tissue,kidney tissue vitamin D receptor (VDR),kidney tissue 1α-hydroxylase (CYP27B1) expression.Western blot assay ventricular muscle tissue,kidney tissue and kidney tissue CYP27B1 and VDR protein relative expression.[Results] Guanmaitong tablets whole components can reduce the Scr significantly (P<0.05),increase T3,T4 effect significantly (P<0.05) and increase serum 1,25(OH)₂D₃ level (P<0.05).The whole components group can increase the expression of CPY27B1 in renal tissue.The whole components and its part components can also increase VDR expression in cardiac and kidney and renal tissues.[Conclusion] Guanmaitong tablets may act on CPY27B1 and VDR as a core part of its mechanism for invigorating the kidney and activating blood circulation action to provide data supporting basic research.