脾气虚与鱼藤酮损伤大鼠模型心肌AgⅡ、BNP及其受体表达差异对比研究

Spleen Qi deficiency and rotenone induced rat model of myocardial Ag Ⅱ,BNP and its receptor expression in the comparative study

目的研究观察脾气虚模型大鼠与鱼藤酮损伤模型大鼠心肌血管紧张素Ⅱ(AgⅡ)、脑钠肽(BNP)及其受体表达差异,探讨鱼藤酮损伤线粒体模型与脾气虚证大鼠模型之间信号转导通路的相关性。方法 SPF级SD雄性大鼠共30只,按照随机数字表随机分为5组,分别是正常组、脾气虚组、鱼藤酮高剂量组、鱼藤酮中剂量组、鱼藤酮低剂量组。正常对照组不做处理,脾气虚组通过饮食不节和力竭游泳的方法建立模型,造模共计15d。鱼藤酮高、中、低剂量组分别通过注射2.0、1.5、1.0ml/kg体重的无菌鱼藤酮油进行干预,1次/d,4周。造模结束后,取大鼠心肌组织,Elisa方法检测大鼠心肌中AgⅡ、BNP含量。Western blot方法检测AgⅡ受体和BNP受体的蛋白表达。免疫组化检测AgⅡ和BNP含量表达。HE染色对心肌组织形态进行观察。结果与正常组相比,脾气虚组AgⅡ含量显著升高(P<0.05),BNP含量显著降低(P<0.05);与脾气虚组相比,鱼藤酮组各组AgⅡ含量均无显著性差异(P>0.05),中剂量组和高剂量组BNP含量无显著性差异(P>0.05)。与正常组相比,脾气虚组AgⅡ受体蛋白含量表达显著升高,差异具有统计学意义(P<0.05),与脾气虚组相比,鱼藤酮组AgⅡ受体蛋白含量表达具有明显变化,但是高剂量组与脾气虚组蛋白表达最为相近;与正常组相比,脾气虚组BNP受体蛋白表达显著降低,差异具有统计学意义(P<0.05),与脾气虚组相比,鱼藤酮高剂量组蛋白表达无显著性差异(P<0.05)。结论用鱼藤酮干预线粒体工作环境方法复制脾虚证模型大鼠与正常脾气虚模型大鼠心肌组织AgⅡ、BNP及其受体表达有关联,为鱼藤酮复制脾气虚证候模型大鼠提供实验依据。

Objective Study of spleen Qi deficiency model rats and rotenone model rat myocardial angiotensin Ⅱ（ Ag Ⅱ）,brain natriuretic peptide（ BNP） and its receptor expression differences,to investigate the correlation between the model of rotenone mitochondrial damage in spleen Qi deficiency rats model and signal transduction pathway. Methods A total of 30 male SPF SD rats,according to the random number table were randomly divided into 5 groups,normal group,spleen Qi deficiency group,high dose of rotenone group,rotenone dose group,rotenone low dose group. The normal control group without treatment,spleen deficiency group through diet and exhaustive swimming model modeling method,a total of 15 days. Rotenone is high,medium and low dose group respectively by injection of 2. 0,1. 5,1. 0 ml/kg body weight of sterile rotenone oil intervention,1 times/day for 4weeks. After the model was finished,the myocardial tissue of rats was taken,and the contents of Ag II and BNP were detected by Elisa method. The expression of Western blot method for detection of Ag II receptor and BNP receptor protein. The expression of Ag II and BNP content was observed by immunohistochemistry. The morphology of cardiac muscle was observed by HE staining.Results Compared with the normal group,spleen Qi deficiency Group Ag II content increased significantly（ P 〈 0. 05）,BNP levels were significantly decreased（ P 〈 0. 05）; and spleen deficiency group compared,rotenone group Ag II content had no significant difference（ P 〉 0. 05）,middle dose group and high dose group of BNP content no significant difference（ P 〉 0. 05）. Compared with the normal group,the temper deficiency group Ag II receptor protein expression was significantly increased,the difference is statistically significant（ P 〈 0. 05）,and spleen Qi deficiency group compared,rotenone group Ag II receptor protein content expression has obvious changes,but the high dose group and spleen Qi deficiency group protein expression which is most similar; compared with the normal group,temper deficiency group BNP receptor protein expression decreased significantly,the difference is statistically significant（ P 〈 0. 05）,and spleen Qi deficiency group compared,rotenone high dose group protein expression had no significant difference（ P 〈 0. 05）. Conclusion With rotenone intervention method in the working environment of the mitochondrial replication model of spleen Qi deficiency rats and normal spleen Qi deficiency model rats myocardial tissue Ag II,BNP and its receptor expression is associated,as rotenone replication of spleen Qi deficiency syndrome model rats to provide experimental basis.