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目的观察AM12质粒(含HBV3.2kb全基因组)对体外培养的人近端肾小管上皮细胞表型转化的作用。方法以体外培养的人近端肾小管上皮细胞系(HK-2)为对象,以AM12质粒转染HK-2细胞为转染组,以正常HK-2细胞作为阴性对照组,采用酶联免疫吸附法(ELISA)检测上清液中转化生长因子-β1(TGF-β1)的浓度。用免疫组织化学检测角蛋白(CK)和α-平滑肌肌动蛋白(α-SMA)表达。结果转染组TGF-β1为(283.85±61.12)pg/ml,高于对照组(210.28±47.21pg/ml)(P<0.05),肾小管上皮细胞免疫组化染色显示CK减弱,α-SMA增强。结论HBV-DNA质粒转染人肾小管上皮细胞可引起TGF-β1分泌增加,并导致肾小管上皮细胞转分化。
Objective To observe the effect of AM12 plasmid (including HBV3.2kb whole genome) on phenotypic transformation of human proximal tubular epithelial cells cultured in vitro. Methods Human proximal tubular epithelial cell line (HK-2) was cultured in vitro. HK-2 cells were transfected with AM12 plasmid, and normal HK-2 cells were used as negative control group. Enzyme-linked immunosorbent assay The concentration of transforming growth factor-β1 (TGF-β1) in the supernatant was detected by ELISA. The expression of keratin (CK) and α-smooth muscle actin (α-SMA) was detected by immunohistochemistry. Results TGF-β1 in the transfected group was (283.85 ± 61.12) pg / ml higher than that in the control group (210.28 ± 47.21 pg / ml) (P <0.05). Immunohistochemical staining of renal tubular epithelial cells showed that CK decreased, Enhanced. Conclusion HBV-DNA plasmid transfected human renal tubular epithelial cells can cause increased TGF-β1 secretion, and lead to renal tubular epithelial cells transdifferentiation.