摘要
目的探讨HBV X基因表达的蛋白(HBVx)对体外培养的大鼠系膜细胞表达肿瘤坏死因子(TNF)-α、白介素(IL)-1β、IL-6及细胞增殖的影响。方法构建含HBV X基因的真核表达载体pCI-neo-X,采用脂质体法将pCI-neo-X转染给体外培养的大鼠系膜细胞,Western blot测定HBx的表达,以半定量逆转录-聚合酶链反应(RT-PCR)测定体外培养大鼠系膜细胞TNF-α、IL-1β、IL-6 mRNA表达,酶联免疫吸附试验(ELISA)测定培养上清液中TNF-α、IL-1β、IL-6水平,四甲基偶氮唑盐(MTT)法测定细胞增殖。结果转染pCI-neo-X的系膜细胞(MC+pCI-neo-X组),HBx在36、48 h表达明显。同时TNF-α、IL-1β、IL-6 mRNA表达量较未转染和转染空载体组明显增高。上清液中TNF-α、IL-1β、IL-6水平在MC+pCI-neo-X组较未转染和转染空载体组均增高,差异均有统计学意义(P<0.05);细胞增殖在36、48 h MC+pCI-neo-X组最明显,差异有统计学意义(P<0.05)。结论 HBx可诱导大鼠系膜细胞高表达TNF-α、IL-1β和IL-6,促进系膜细胞增殖。HBx对系膜细胞的增殖作用可能与TNF-α、IL-1β、IL-6的高表达有关。
Objective To approach the effect of HBx on expression of tumor necrosis factor-α（TNF-α）,interleukin（IL）-1β and IL-6,and proliferation of mesangial cell of rat in vitro.Methods Liposome bearing pCI-neo-X was transfected into cultured rat mesangial cell line in vitro through liposome method.X gene expression in the cells was assessed by Western blot.The mRNA expressions of TNF-α,IL-1β and IL-6 in cultured mesangial cells line were assessed by semiquantitative RT-PCR.The levels of TNF-α,IL-1β and IL-6 were measured by ELISA.Mesangial cell proliferation was assessed by MTT.Results HBx obviously expressed in cultured mesangial cells line at 36th and at 48th hour after transfection.The mRNA of TNF-α,IL-1β and IL-6 expression simultanously increased.The supernant TNF-α,IL-1β and IL-6 levels in pCI-neo-X group were all markedly higher than those in other two groups（all P0.05）.The cell proliferation was also obvious at the same time（all P0.05）.Conclusion Therefore,we can conclude that HBx can induce expression of TNF-α,IL-1β and IL-6.HBx may play a critical role in the cell proliferation through TNF-α,IL-1β and IL-6 high expression.
出处
《重庆医学》
CAS
CSCD
北大核心
2011年第22期2227-2230,共4页
Chongqing medicine
基金
国家自然科学基金资助项目(30772360)
湖北省卫生厅计划课题(JX4B47)
长江大学博士启动基金资助项目(2009001)
