乙型肝炎病毒X基因转染大鼠系膜细胞对细胞增殖及IL-1β,IL-6表达的影响  

HBV X gene transfection upregulates IL-1β and IL-6 gene expression and induces rat glomerular mesangial cell proliferation

作　　者：卢宏柱[1] 樊启红[2] 周建华[3] 

机构地区：[1]长江大学临床医学院儿科教研室,湖北荆州434000 [2]长江大学附属第一医院儿科,湖北荆州434000 [3]华中科技大学同济医学院附属同济医院儿科,武汉430030

出　　处：《广东医学》2009年第11期1611-1614,共4页Guangdong Medical Journal

基　　金：国家自然科学基金课题(编号:30772360)

摘　　要：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(IL)-1β、IL-6及细胞增殖的影响。方法将HBVX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-1β mRNA、IL-6 mRNA表达;MTT法测定细胞增殖。结果转染PCI-neo-X的系膜细胞,HBx在36和48h表达明显。同时IL-1β mRNA、IL-6 mRNA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48h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-1β mRNA和IL-6 mRNA,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。The HBV X gene encodes a 17 -kd protein, termed HBx, that has been shown to function as a transcriptional trans - activator of a variety of viral and cellular promoter/enhancer elements. This study was aimed to investigate 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 endonuclease digestion and sequencial analysis. PCI - neo - X was transfected into cultured rat mesangial cell line in vitro via liposome. HBx expression in transfeeted mesangial 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 significantly expressed in euhured mesangial cell line at 36 and 48 hours after transfeetion. 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 tranfeetion can induce IL - 1β and IL - 6 expression and mesangial cell proliferation. HBx may play a critical role in mesangial cell proliferation through upregulation of IL - 1β and IL - 6 expression.

关 键 词：白细胞介素-1Β 白细胞介素-6 乙型肝炎病毒X基因 系膜细胞 大鼠 

分 类 号：R714.502[医药卫生—妇产科学] R446.61[医药卫生—临床医学]