黄连解毒汤对自发性高血压大鼠主动脉组织miR-133a/Caveolin-1/eNOS通路调控作用机制的研究

Study of Huanglian Jiedu Decoction on the regulation mechanism of the aortic tissue miR-133a/Caveolin-1/eNOS pathway in spontaneously hypertensive rats

目的:探讨黄连解毒汤对自发性高血压大鼠(SHR)主动脉组织miR-133a/Caveolin-1/eNOS通路关键因子表达的影响。方法:SHR随机分为模型组,卡托普利组,黄连解毒汤低、中、高剂量组,WKY大鼠作为正常组,每组各10只。给予相应药物灌胃后,检测各组大鼠血清NO、内皮素-1(ET-1)及主动脉eNOS变化,RTPCR检测大鼠主动脉组织miR133a、Caveolin-1、eNOS mRNA表达,Western Blot检测大鼠主动脉组织Caveolin-1、eNOS、p-eNOS蛋白表达。结果:与正常组比较,模型组大鼠血清NO、主动脉eNOS含量显著降低(P<0.01),血清ET-1含量显著升高(P<0.01),模型组大鼠主动脉miR-133a、eNOS mRNA表达显著降低(P<0.01),主动脉Caveolin-1 mRNA和蛋白表达显著升高(P<0.01),主动脉eNOS、p-eNOS蛋白表达显著降低(P<0.01)。与模型组比较,各治疗组大鼠血清中NO、主动脉eNOS的含量不同程度升高,血清ET-1的含量不同程度降低。卡托普利组及黄连解毒汤中、高剂量组大鼠主动脉miR-133a、eNOS mRNA表达显著升高(P<0.01),Caveolin-1 mRNA和蛋白表达显著降低(P<0.01),eNOS、p-eNOS表达水平显著升高(P<0.01)。结论:黄连解毒汤可通过升高SHR主动脉miR-133a、eNOS mRNA表达水平及eNOS、p-eNOS蛋白的表达水平,降低Caveolin-1 mRNA及蛋白表达水平保护SHR大鼠血管内皮功能,达到降低血压的作用。

Objective:To investigate the effects of Huanglian Jiedu Decoction(HLJDD)on the expression of key molecules of miR-133a/caveolin-1/eNOS pathway in the aorta of spontaneously hypertensive rats(SHR).Methods:SHR was randomly divided into model group,captopril group,HLJDD low,medium and high dose group,WKY rats as normal group,10 rats in each group.After gavage of corresponding drugs,the changes of serum NO,ET-1 and aortic eNOS were detected.The m RNA expressions of miR133a,Caveolin-1 and eNOS were detected by RT-PCR,the protein expressions of Caveolin-1,eNOS and p-eNOS were detected by Western Blot.Results:Compared with the control group,the model group’s levels of NO in serum and eNOS in aorta decreased significantly(P<0.01),the levels of ET-1 in serum increased significantly(P<0.01),the expression of miR-133a and eNOS mRNA decreased significantly(P<0.01),and the expression of Caveolin-1 mRNA and protein increased significantly(P<0.01),and the expression of eNOS and p-eNOS protein decreased significantly(P<0.01).Compared with the model group,the levels of NO and eNOS in serum and aorta were significantly increased in each treatment group,the levels of ET-1 in serum decreased significantly,the expressions of miR-133a and eNOS mRNA in aorta of rats in captopril group,HLJDD medium and high dose group were significantly higher(P<0.01),and the expression of Caveolin-1 mRNA and protein in aorta of rats in captopril group,HLJDD medium and high dose group was significantly lower(P<0.01),and the expressions of eNOS and p-eNOS in aorta in captopril group,HLJDD medium and high dose group were significantly higher(P<0.01).Conclusion:HLJDD can increasing the expression levels of miR-133a,eNOS mRNA,eNOS,p-eNOS protein expression,decreased the Caveolin-1 mRNA and protein expression,while protect vascular endothelial function and lower blood pressure of SHR.