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目的以流感病毒感染的小鼠为模型,检测犀角地黄汤合银翘(XDY)对小鼠肺组织中肌球蛋白轻链(MLC)磷酸化信号通路的影响,以明确其抗病毒作用的分子机制。方法将50只雄性Balb/c小鼠按体重随机分为5组,每组10只,分别为正常组、病毒组和XDY低、中、高剂量给药组,除正常组小鼠外,其余组用A/FM/1/34(H1N1)病毒株滴鼻感染。1 h后,病毒组蒸馏纯水灌胃,2次/d;其余三组分别用低、中、高剂量的XDY灌胃,2次/d。感染后第4天摘眼球处死小鼠,RT-PCR检测各组小鼠肺组织中病毒RNA水平;Western bolt检测肺组织MLC、p-MLC、PKC、p-PKC、ROCK-1、ERM、p-ERM蛋白表达水平。结果与病毒组相比,XDY中剂量组病毒RNA含量显著降低,差异有统计学意义(P<0.01),而高剂量组和低剂量组病毒RNA含量则无明显变化。XDY中剂量组Rho/ROCK、PKC以及p-MLC、p-ERM蛋白表达显著降低,差异有统计学意义(P<0.05)。与正常组相比,病毒组小鼠肺组织中ROCK1、PKC表达显著增加,p-MLC、p-ERM蛋白表达显著升高,差异有统计学意义(P<0.05)。结论 XDY可通过抑制Rho/ROCK、PKC信号通路活化,抑制p-MLC蛋白表达,减少流感病毒在体内的复制,从而起到抗病毒作用。
Objective To investigate the effect of XDH on the phosphorylation of myosin light chain (MLC) in mouse lung tissue by using influenza virus infected mice as a model to detect the anti-viral molecules mechanism. Methods Fifty male Balb / c mice were randomly divided into five groups according to body weight, with 10 mice in each group. The mice were divided into normal group, virus group and XDY low, medium and high dose groups. Except normal mice, Group Infected with A / FM / 1/34 (H1N1) strain. After 1 h, the virus group was distilled water for 2 times / d. The other three groups were administered with low, medium and high doses of XDY twice daily. The mice were sacrificed on day 4 after infection, and the levels of viral RNA in lung tissue were detected by RT-PCR. The expressions of MLC, p-MLC, PKC, p-PKC, ROCK- -ERM protein expression level. Results Compared with the virus group, the content of RNA in the middle-dose XDY group was significantly lower (P <0.01), while there was no significant change in the RNA content in the high-dose and low-dose groups. The expression of Rho / ROCK, PKC, p-MLC and p-ERM in XDY middle dose group was significantly decreased (P <0.05). Compared with the normal group, the expression of ROCK1, PKC and p-MLC, p-ERM in the lung tissue of the virus group were significantly increased, the difference was statistically significant (P <0.05). Conclusion XDY can play an antiviral effect by inhibiting the activation of Rho / ROCK and PKC signaling pathway, inhibiting the expression of p-MLC protein and reducing the replication of influenza virus in vivo.