HBV-DNA阳性血清对足细胞增殖、凋亡及其细胞周期调控蛋白mRNA表达的影响

目的：探讨人类乙肝病毒DNA阳性血清对体外培养小鼠足细胞增殖、凋亡及cyclin D1及p27mRNA表达的影响,并探讨其意义.方法：以体外培养的小鼠肾小球足细胞为对象,分别用健康人血清（C组）、携带HBV无肾损害者阳性血清（B组）和乙型肝炎病毒相关性肾炎（HBV-GN）者HBV-DNA阳性血清 依据其HBV-DNA含量分为A1组（〈10^3 copy/ml）、A2组（10^4 copy/ml）及A3组（10^6 copy/ml）] 刺激,采用MTT检测足细胞增殖水平,Annexin V法检测足细胞凋亡,RT-PCR法检测足细胞cyclin D1及p27mRNA的表达水平.结果：人类HBV-DNA阳性血清对小鼠分化足细胞有明显刺激增生作用,与C组比较,A1组对足细胞增殖无明显影响,而A2、A3组及B组血清均能显著刺激足细胞发生增殖,但各组之间比较差异无统计学意义.HBV-DNA阳性血清对小鼠足细胞的凋亡无明显影响.与空白组及C组比较,A1、A2、A3组和B组阳性血清对小鼠足细胞cyclin D1mRNA的表达无明显影响,但可明显下调足细胞p27mRNA的表达,但各组之间差异无统计学意义.结论：人类HBV-DNA阳性血清可刺激小鼠分化足细胞发生增殖,但对细胞中cyclin D1的表达和凋亡没有影响,其刺激增殖作用可能与下调p27mRNA的表达有关.

The Effect of Human HBV-DNA Positive Serum on Cell Proliferation and Apoptosis and the Expression of p27 mRNA,Cyclin D1mRNA of Mouse Podocyte

Objective：To investigate the effect of human HBV - DNA positive serum on cell Proliferation, cell Apoptosis and the expression of p27 mRNA, cyclin D1 mRNA of mouse podocyte, and explore their clinical value in HBV- GN. Methods ：The mouse podocyte was cultured in the presence of the human serum of healthy person （ group C） and the HBV - DNA positive serum from patients without kidney disease. （ group B ： 106 copy/ml） or with HBV - GN （ Group A1 ： HBV - DNA 〈 103 copy/ml ; A2 ： 104 copy/ml and Group A3：106 copy/m]）. The proliferation measurement of each group was determined by MTF, the apoptosis measurement by Annexin and the mRNA measurement of p27mRNA and cyclinDlmRNA by RT- PCR. Resulls： Human HBV -DNA positive serum could stimulate podocyte proliferation. Compared with group C, no cell proliferation of podocyte was found in group A1, while HBV - DNA positive serum in group B and group A2 and A3 could significantly stimulate mouse podocyte proliferation. There was no signifi- cantly difference in. the later three groups. No obvious effect of human HBV - DNA positive serum on the apoptosis of podocyte was found in all groups. Compared with control group and group C , Human HBV positive serum in group B ,Group AI ,group A2 and group A3 had no significant effect on the expression of cyclin D1 mRNA of podocyte, but the p27mRNA was significantly down -regu- lated. Conclusion： Human HBV - DNA positive serum can stimulate mouse podocyte to proliferate, but has no significantly effect on its apoptosis and the expression of cyclin DlmRNA. The proliferation of podocyte may be attributable to the down - regulation of the expression of p27.