从miR-133a/Caveolin-1/eNOS通路研究“黄连解毒汤上调SHR eNOS/NO”

高血压是重大公共卫生问题，清热解毒是中医防治高血压的重要治法之一，黄连解毒汤（HLJDD）是清热解毒的古典经方。申请者前期研究表明HLJDD对SHR主动脉的eNOS/NO/PKGⅠ信号上调、miR-133a过度表达，miR-133a高表达可使内皮细胞Caveolin-1下调，Caveolin-1 又负调控eNOS。推测HLJDD对SHR高表达的mi-133a使 Caveolin-1下调，并升高eNOS含量及活性。本项目通过HLJDD干预SHR，采用荧光定量PCR、Western blot检测主动脉mir-133a、caveolin-1、p-eNOS表达及eNOS活性，并以HLJDD含药血清干预牛主动脉内皮细胞，检测相应mRNA和蛋白表达，探讨miR-133a/Caveolin-1/eNOS通路在“HLJDD上调SHR主动脉eNOS/NO信号”作用机制，为清热解毒方临床应用提供实验支持。

Hypertension is an important public health problem, Qing Re Jie Du is one of the important Chinese medical treatment, and the decoction of Huang Lian Jie Du is a classic ancient formula. The applicants previously found that HLJDD had the effect of up-regulating the signals of eNOS/NO/ PKG and causing the over-expression of miR-133a, and the over-expression of miR-133a could cause the down-regulating endothelial cells’ Caveolin-1 and negative regulating the eNOS. In this project, we hypothesized that HLJDD up-regulating miR-133a decrease Caveolin-1 is probably the mechanism of up-regulating the content and the activity of eNOS. This project will treat the SHR with HLJDD and using Real-time Quantitative PCR and Western blot to detect the RNA and protein expression of mir-133a, caveolin-1，eNOS, and the activity of eNOS, and same experiments will do in vitro. This project will illuminate that the miR-133a/ Caveolin-1/eNOS path-way weather it involve the mechanism of the HLJDD up-regulating the eNOS/ NO signal of SHR aorta or not. The results will provide new evidences for the clinic usage of the ancient classic formula HLJDD.