The HBV X gene encodes a 17 -kd protein, termed HBx, that has been shown to function as a tran-scriptional trans -activator of a variety of viral and cellular promoter/enhancer elements. This study was aimed to investi-gate the effect of HBx on gene expression of interluekin (IL) -1β and IL -6, and proliferation of rat mesangial cell in vitro. The HBV X gene was amplified with PCR assay, and inserted into the eukaryotic expression vector PCI -neo. The structure of recombinant PCI -neo -X plasmid was confirmed by restrict endonuelease digestion and sequencial analysis. PCI -neo -X was transfected into cultured rat mesangial cell line in vitro via liposome. HBx expression in transfected me-sangial cells was assessed by Western blot. The IL -1β and IL-6 mRNA expression in those cells was assessed by semi -quantitative RT -PCR. Mesangial cell proliferation was assessed by MTT. The results showed that HBx was significant-ly expressed in cultured mesangial cell line at 36 and 48 hours after transfection. The IL -1β and IL -6 mRNA levels were simutanously increased. The cell proliferation was also elevated at the same time compared to the control groups. Therefore, it can be concluded that HBx gene tranfection can induce IL -1β and IL -6 expression and mesangial cell prolif-eration. HBx may play a critical role in mesangial cell proliferation through upregulation of IL -1β and IL-6 expression.%目的 探讨乙型肝炎病毒(HBV)X基因表达的蛋白(HBx)对体外培养的大鼠系膜细胞表达白细胞介素(1L)-1β、IL-6及细胞增殖的影响.方法 将HBV X基因扩增,与PCI-neo载体连接,构成PCI-neo-X真核表达载体.采用脂质体法将PCI-neo-X转染给大鼠系膜细胞,Western blot测定HBx的表达;以半定量反转录PCR测定体外培养大鼠系膜细胞IL-1βmRNA、IL-6mRNA袁达;MTT法测定细胞增殖.结果 转染PCI-neo-X的系膜细胞,HBx在36和48 h表达明显.同时IL-1βmRNA、IL-6mRNA表达量较未转染和转染空载体者明显增高.细胞增殖在36和48 h最明显,与未转染和转染空载体组有显著差异.结论 HBx可诱导大鼠系膜细胞高表达IL-1βmRNA和IL-6mRNA,促进系膜细胞增殖.HBx对系膜细胞的增殖作用可能与IL-1β、IL-6的高表达有关.
展开▼
