基于心肌能量和功能、心脏微循环动态和微血管通透性的调控，探讨芪参益气滴丸补气行血和补气摄血的机理

Cardiac ischemia and reperfusion(I/R)injury after interventional therapy for acute cardiac infarction remains a challenge in clinic. Ischemia-induced diminution in cardiomyocyte energy and function falls into the deficiency of qi. The decrease in red blood cell velocity in cardiac microvessels and cardiac perfusion after reperfusion belongs to the inability of qi to move blood, while albumin leakage from venules belongs to inability of qi to conserve body fluid. To tonify qi to move blood and tonify qi to conserve blood is the widely used management in Chinese medicine for the diseases that are associated with the inability of qi to move blood and conserve body fluid. Qishenyiqi pills (QSYQ) is a compound preparation of Chinese medicine based on this principal. Clinic and basic researches both reveal its efficiency in attenuation of I/R-induced cardiac injury. However, it is not clear at present what is the mechanism in terms of tonifying qi to move and conserve blood. The present work intends to clarify the mechanism for tonifying qi of QSYQ in a rat I/R model, using a dynamic visualized technique in conjunction with morphology, molecule biology, and analysis of cardiac energy, structure and function; the mechanism for tonifying qi to move blood by observation of the dynamics in cardiac microcirculation; and the mechanism for tonifying qi to conserve blood by assessment of cardiac microvascular permeability, the regulation of aquaporin-1, caveolae, tight junction proteins in cardiac microvascular endothelium, in the hope to sublimate the theory of Chinese medicine.

急性心肌梗死介入治疗引发的心肌缺血再灌注(I/R)损伤是尚未解决的临床问题。缺血引起的心肌能量和心功能的降低属于心气虚范畴。再灌注后出现的心脏微血管内红细胞流速和心脏灌流量的降低属于气不行血的范畴；出现的血浆白蛋白经由细静脉的外漏属于气不摄津液的范畴。补气行血和补气固摄是中医治疗气不行血和气不固摄相关疾病的常用治法，芪参益气滴丸(QSYQ)是依据该法组成的复方制剂，临床和基础研究已经明确了其有改善I/R诱导的心肌损伤的作用，但是，其补气行血和补气固摄作用机理尚不清楚。本研究用心肌缺血和I/R损伤的大鼠模型，用动态可视化技术，结合形态学观察和分子生物分析，通过心肌能量、结构、功能的调控，阐明其补心气的机理；通过心脏微循环动态的调控，阐明其补气行血的机理；通过心脏微血管通透性、微血管内皮细胞水通道-1、脂膜微囊、细胞缝隙连接蛋白的调控，阐明其补气固摄的机理，借以升华中医理论。

QSYQ 及其主要成分黄芪甲苷(AS IV)、人参皂苷Rb1(Rb1)、Rg1、三七皂苷R1(R1)、丹参素 (DLA) 均能改善心肌缺血大鼠心肌结构损伤。QSYQ、AS IV可抑制大鼠心肌ATP/ADP、ATP/AMP比值的降低，抑制血浆中cTnI含量的升高。.QSYQ可以抑制I/R引起的大鼠心肌ATP5D低表达，增加心肌ATP含量，改善心肌F-actin，心肌结构，心功能和心灌流量，减少心肌梗死。AS IV、Rb1、Rg1、R1均可抑制大鼠心肌ATP5D低表达，改善心肌能量代谢，改善心肌结构，心功能和心灌流量，减少心肌梗死。揭示了QSYQ及其主要补气成分补气行血的作用机理。QSYQ主要活血成分DLA通过与Sirt-1的结合，抑制I/R引起的大鼠心肌NDUFA 10蛋白和mRNA的低表达，改善Complex Ⅰ的表达和活性，抑制氧化应激损伤，减轻心肌梗死。.QSYQ及其补气成分均可抑制Rac-1表达的升高。除Rb1外QSYQ及其他成分均可抑制PAK-1表达的下降。QSYQ、AS IV、DLA可以抑制PKCα的低表达。 QSYQ及其成分均能抑制ERα低表达。QSYQ抑制了I/R引起的大鼠冠状血管细静脉血浆白蛋白外漏和血管内皮细胞连接的开放。QSYQ及AS IV、Rg1、Rb1、R1可以抑制缺氧复氧引起的大鼠冠状血管内皮细胞的ATP/AMP比值的降低， F-actin断裂和重排，抑制血管内皮细胞连接蛋白Claudin-5、Occludin、ZO-1的排列紊乱， p-VE-cadherin、p-Src、p-Caveolin-1的升高。QSYQ及其成分均能抑制Rac-1、ROCK及MMP-2表达的升高；QSYQ、ASIV、Rb1可以抑制HR细胞中PKCa的降低；QSYQ、AS IV、Rb1、DLA可以抑制HR细胞中MMP-9的升高。阐明了QSYQ补气固摄的机理。