【摘要】 通过构建A型流感病毒H1N1感染小鼠模型,并使用c-Jun蛋白的特异性核酶Dz13抑制该蛋白的表达,探索c-Jun蛋白在流感病毒感染后对T淋巴细胞增殖和炎性反应的调节作用。蛋白质印迹法(Western-blotting)结果表明,核酶Dz13作用后c-Jun蛋白的表达量明显降低,且c-Jun蛋白的活化也随之降低。此外,用流式细胞术测定感染后3 d和6 d外周血内辅助性T细胞(CD4~+T)和杀伤性T细胞(CD8~+T)的增殖情况,同时用实时荧光定量聚合酶链式反应测定外周血内细胞因子IFN-γ、IL-6和IL-10表达。结果显示:小鼠感染病毒后,外周血CD4+和CD8+T细胞明显增多,相关细胞因子的表达量也明显上调;而抑制c-Jun蛋白表达后,CD4~+和CD8~+T细胞的增殖受到抑制,且相关细胞因子IFN-γ、IL-6和IL-10的表达量亦受到抑制。说明c-Jun蛋白参与调节A型流感病毒H1N1感染后引起的T淋巴细胞增殖和相关细胞因子的表达。更多还原

【Abstract】 A H1N1 influenza A virus infected mice model was built to evaluate the regulation mechanism of c-Jun protein in T lymphocytes. To determine the DNAzyme Dz13 efficiency in virus-infected animals, a Westernblotting assay was employed to detected total c-Jun(t-c-Jun) and phosphorylation c-Jun(p-c-Jun) levels by specific antibody. It was demonstrated that the drug-treated mice markedly suppressed the expression of t-c-Jun and p-c-Jun. Through flow cytometry and quantitative real-time polymerase chain reaction(qRT-PCR), we could find an obvious CD4~+(helper T lymphocytes) and CD8~+T(cytotoxic T lymphocyte) cell proliferation and relative cytokine(IFN-γ, IL-6 and IL-10) expression on day 3 and 6 in the virus-infected mice. The Dz13-treated mice with a c-Jun suppression displayed a down-regulation of CD4+and CD8+T cells in peripheral blood. At the same time, Dz13-treated group demonstrated a significant reduction of inflammatory cytokine(IFN-γ, IL-6 and IL-10) expression. In conclusion, c-Jun protein regulates CD4~+ and CD8~+T cell proliferation and inflammatory cytokine expression in H1N1 influenza A virus infected mice.更多还原