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目的:探讨流感病毒H1N1体外感染细胞诱导炎性因子表达及黄芩苷调控作用的研究。方法:正常生长细胞接种100 TCID50的病毒液;吸附2h后加入黄芩苷高剂量组(3.96μg/m L)和黄芩苷低剂量组(0.99μg/m L),以奥司他韦(0.75μg/m L)做阳性对照。采用基因芯片技术分析各组细胞差异表达炎性因子。以RT-PCR和Western blotting法检测细胞内靶因子m RNA和蛋白的表达变化。结果:与细胞对照组相比,H1N1感染组差异表达基因IL-1β、IL-6、IL-8、TNF-α和RANTES明显上调(P<0.01);给药后上述因子表达下调,部分IL-10明显上调(P<0.05,P<0.01)。Western blotting和RT-PCR检验结果与基因芯片结果相符合。结论:流感病毒H1N1感染后诱导炎性因子过度表达损伤细胞,黄芩苷可以调控炎性因子表达,减轻宿主细胞的损伤作用,具有明显的抗感染在用。
Objective: To investigate the expression of inflammatory cytokines induced by influenza virus H1N1 in vitro and the regulation of baicalin. Methods: Normal cells were inoculated with 100 TCID50 virus solution. After 2 h of adsorption, baicalin high dose (3.96 μg / m L) and baicalin low dose (0.99 μg / m L) / m L) as a positive control. Gene chips were used to analyze the differentially expressed inflammatory cytokines in each group. RT-PCR and Western blotting were used to detect the expression of m RNA and protein. Results: The expression of IL-1β, IL-6, IL-8, TNF-α and RANTES were significantly up-regulated in H1N1 infection group compared with the cell control group (P <0.01) -10 was significantly increased (P <0.05, P <0.01). The result of Western blotting and RT-PCR was consistent with the result of gene chip. CONCLUSION: Influenza virus H1N1 induces over-expression of inflammatory cytokines in infected cells. Baicalin can modulate the expression of inflammatory cytokines and attenuate the damage of host cells, with obvious anti-infective effect.