AM12质粒对体外肾小管上皮细胞凋亡和分泌转化生长因子-β_1的影响

Effect of AM12 plasmid on apoptosis and TGF-β_1 secretion of renal tubular epithelial cell in vitro

目的观察AM12质粒(含乙型肝炎病毒3.2 kb全基因组)对体外培养的人近端肾小管上皮细胞HK-2凋亡和分泌转化生长因子-β1(TGF-β1)的影响。方法以体外培养的HK-2细胞为对象,以AM12质粒转染HK-2细胞为转染组,以正常HK-2细胞作为对照组,用流式细胞仪检测细胞的凋亡及Fas表达率;用酶联免疫吸附法(ELISA)检测上清液中TGF-β1的浓度。结果转染组HK-2凋亡率为(1.49±0.02)%,高于对照组的(1.03±0.09)%(P<0.05);Fas表达率为(10.09±2.34)%,高于对照组的(6.58±0.65)%(P<0.05);上清液TGF-β1水平为(283.85±61.12)ng.L-1,高于对照组的(210.28±47.21)ng.L-1(P<0.05)。结论 HBV-DNA质粒转染体外培养的人肾小管上皮细胞可诱导其凋亡并上调其对TGF-β1的分泌。

Objective To investigate the effects of AM12 plasmid（eontain hepatitis B virus 3.2 kb whole genome） on apoptosis and transforming growth factor-renal tubular epithelial cell ; hepatitis B virus ; transfection; apoptosis ; transforming growth factor-β1 （TGF-renal tubular epithelial cell ; hepatitis B virus ; transfection; apoptosis ; transforming growth factor-β1） secretion of renal tubular epithelial cells in vitro. Methods The HK-2 cells were cultured in vitro and then were transfected by AM12 plasmid as transfection group. The normal HK-2 cells were as control group. The apoptosis and the expression rate of Fas were detected by flow cytometry. The TGF-I31 levels in the supernatant were measured by enzyme linked immunosorbent assay （ELISA）. Results The apoptosis rate of HK-2 cells in transfection group（ 1.49 ± 0.02 ） % was significantly higher than that in control group （ 1.03 ± 0. 09 ） % （ P 〈 0.05 ）. The expression rate of Fas in transfection group （ 10. 09 ± 2.34 ） % was significantly higher than that in control group （ 6.58±0. 65 ） % （ P 〈 0. 05 ）. The TGF-β1 level in transfection group（283.85 ± 61.12） ng· L-1 was significantly higher than that in control group （210.28±47.21 ） ng · L-1 （ p 〈 0.05 ）. Conclusion The HBV-DNA plasmid transfect renal tubular epithelial cells which were cultured in vitro could induce apoptosis of HK-2 and up-regulating the secretion of TGF-β1 of HK-2.