清心饮及其含药血清对培养大鼠心肌细胞感染柯萨奇B_3病毒的保护作用

PROTECTIVE EFFECT OF QINGXINYIN AND ITS PHARMACOLOGIC SERUM ON RAT'S CULTURED CARDIOMYOCYTES FROM THE INFECTION OF COXSACKIC VIRUS B_3

本实验旨在观察清心饮对病毒性心肌炎模型的保护作用,并初步探讨其机制。选用培养SD大鼠心肌细胞感染柯萨奇B_3病毒,作为实验性病毒性心肌炎模型,研究分为两部分,分别采用直接添加药液于细胞培养基中和血清药理学两种不同的离体实验方法。实验中观察了细胞形态、细胞病变及搏动性情况,测定感染后3d及5d细胞上清液中的病毒滴度和心肌酶(包括乳酸脱氢酶、天冬氨酸氨基转移酶),并通过斑点杂交技术观察了清心饮对柯萨奇B_3病毒核糖核酸复制的影响。结果显示:清心饮及其含药血清均可明显减轻细胞病变,改善细胞搏动,降低细胞上清液中的病毒滴度(分别与病毒对照组和空白血清对照组比P<0.05～O.01)。清心饮及其含药血清均可明显减少感染心肌细胞乳酸脱氢酶、天冬氨酸氨基转移酶的释放(分别与病毒对照组和空白血清对照组比P<0.05～0.01),清心饮及其含药血清均可显著抑制柯萨奇B_3病毒核糖核酸的复制(分别与病毒对照组和空白血清对照组比P<0.05～0.01)。由此提示清心饮对病毒性心肌炎具有明显的抗病毒和细胞病变保护作用,抑制病毒核酸的复制可能是其机理之一。血清药理学研究提示清心饮中含抗病毒、保护心肌细胞的有效成分,口服有效。

In order to observe the protective effect of Qing Xin Yin ( QXY) and its serum from viral myocarditis infected, monolayer culture of spontaneously contracting rat heart cells were obtained from newborn SD rats and 100TCID50 of CVB3(Nancy Strain) was seeded into plates after cultured 18hours. The experiment was divided into two parts, and two different test approaches in vitro -the medicinal extract was added to medium directly and serologic pharmacological test were adopted respectively. In experiment, the beating of cells and cytopathic effect were observed, virus titers and cardiac cytosolic enzymes in supernatant after infection were detected, and effect of QXY on replication of CVB3-RNA was investigated by Dot blot technique. The results showed that both serum with QXY and QXY itself could obviously decrease cytopathic effect, improve beating condition of cells, attenuate virus titers in supematant and inhibit the release of cardiac cytosolic enzymes-lactic acid dehydrogenase and aspartale amino transferase (Compared with the virus control and blank serum control respectively. P< 0.05 -0.01). It was also found that both serum with QXY and QXY itself could restrain replieation of CVB3-RNA in myocardium significantly by semi-quantitative analysis (compared with the virus conlrol and blank serum control respectively, P<0.05-0.01) All these indicated that QXY was clearly effective on resisting CVB3 and protecting myocardium on viral myocarditis infected models, and one of its mechanisms might be due to the effect of restraining replication of CVB3-RNA, serologic pharmacological test indicated lhal QXY'S available composition of antivirus and protecting myocardium could be active after taken orally.