从p38MAPK信号通路介导HSP27表达研究平肝潜阳法干预高血压肝阳上亢证的作用机制

As one of the important methods to prevent and treat essential hypertension, the study shows that the method of calming liver and suppressing Yang can reverse the vascular remodeling, but the mechanism has not been clarified. This project uses the classical prescription of calming liver and suppressing Yang method-Tianma Gouteng decoction as a research tool, to reverse the vascular remodeling as a starting point, from the level of clinical, animal and cell, by morphology, immunofluorescence staining, confocal laser and other molecular biological methods, combined with the over expression and down expression technology. There are three aspects to clarify the mechanism in calming liver and suppressing Yang calming method intervene vascular remodeling in essential hypertension with hyperactivity of liver-Yang syndrome through the p38MAPK signal pathway regulating the expression of HSP27: (1) The method of calming liver and suppressing Yang calming could reverse vascular remodeling by involving in regulating the expression of MKK3/MKK6-p38MAPK mechanism. (2) The method of calming liver and suppressing Yang calming could reverse vascular remodeling by involving in regulating HSP27 expression mechanism. (3) The p38MAPK-HSP27 signaling pathway is involved in vascular remodeling mechanism on essential hypertension with hyperactivity of liver-Yang syndrome. The method of calming liver and suppressing Yang treatment on essential hypertension with hyperactivity of liver-Yang syndrome is illustrated through the mechanism of MKK3/MKK6-p38MAPK-HSP27 signaling pathway. These results can help to clarify the scientific connotation of calming liver and suppressing Yang method on treatment of essential hypertension with hyperactivity of liver-Yang syndrome. The findings suggest that it can provide some experimental basis for clinical prevention and treatment of essential hypertension.

平肝潜阳法作为中医防治高血压的重要治则之一，研究表明能够逆转血管重构，但机制尚未阐明。本项目采用平肝潜阳法经典方剂-天麻钩藤饮为研究工具，以逆转血管重构为切入点，从临床、动物和细胞水平；运用形态学、免疫荧光染色、激光共聚焦和分子生物学方法，结合过表达和抑制表达技术；从3个方面阐明p38MAPK信号通路介导HSP27表达参与平肝潜阳法逆转高血压肝阳上亢证血管重构的作用机制：(1)平肝潜阳法调控MKK3/MKK6-p38MAPK表达参与逆转血管重构的作用机制；(2)平肝潜阳法调节HSP27表达参与逆转血管重构的作用机制；(3)p38MAPK-HSP27信号通路参与高血压肝阳上亢证血管重构的作用机制。揭示平肝潜阳法干预高血压肝阳上亢证是通过调控MKK3/MKK6-p38MAPK-HSP27信号通路的作用机制；有助于阐明平肝潜阳法干预高血压肝阳上亢证的科学内涵，为临床防治高血压提供实验依据。

平肝潜阳法作为中医防治高血压的重要治则之一，研究表明能够逆转血管重构，但机制尚未阐明。本项目采用平肝潜阳法经典方剂-天麻钩藤饮为研究工具，以逆转血管重构为切入点，从临床、动物和细胞水平；运用形态学、免疫荧光染色、激光共聚焦和分子生物学方法，结合过表达和抑制表达技术；从3个方面阐明p38MAPK信号通路介导HSP27表达参与平肝潜阳法逆转高血压肝阳上亢证血管重构的作用机制：(1)从临床资料调查结果显示，高血压病患者不同中医证候在临床方面存在差异性，其中肝阳上亢证主要表现为血压升高、血压变异、节律异常，同时在心室重构、血管重构及血管内皮舒张功能障碍、血管活性物质等方面存在差异性，在外周血HSP27含量及miRNAs表达水平存在差异性。（2）从临床研究结果显示，平肝潜阳法方药能够通过调控外周血HSP27含量及miRNAs表达水平，调节外周血管活性物质水平，改善心室重构、血管重构及血管内皮舒张功能障碍，缓解临床症状。（3）从动物实验证实平肝潜阳法方药通过调控MKK3/MKK6-p38MAPK信号通路，调节HSP27蛋白表达参与逆转血管重构的作用机制；（4）从细胞水平证实平肝潜阳法方药通过抑制miRNA20a表达，激活p38MAPK信号通路，导致增强HSP27表达，促进VSMCs细胞凋亡、抑制细胞增殖、迁移及骨架重排。(5)从细胞水平证实HSP27与MKK3/MKK6-p38MAPK信号通路存在直接相互作用；（6）平肝潜阳方药能够延缓自发性高血压大鼠模型的血管衰老作用，其机制可能与调控主动脉组织中SIRT1-PTEN信号通路表达有关。揭示平肝潜阳法干预高血压肝阳上亢证是通过调控MKK3/MKK6-p38MAPK-HSP27信号通路的作用机制；有助于阐明平肝潜阳法干预高血压肝阳上亢证的科学内涵，为临床防治高血压提供实验依据。