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目的探讨HOXA13对人血清白蛋白(HSA)诱导的人肾小管上皮细胞(HKC)间充质转分化(EMT)和骨形成蛋白7(BMP-7)表达的影响。方法体外培养人肾小管上皮细胞(HKC)经20 mg/ml人血清白蛋白作用48 h后,采用Real-Time PCR和免疫印迹法分别检测人肾小管上皮细胞角蛋白(CK)、波形蛋白(Vimentin)和HOXA13 mRNA和蛋白表达。应用脂质体转染技术上调人肾小管上皮细胞HOXA13表达,观察HOXA13不同表达水平对人血清白蛋白超载诱导CK、Vimentin、BMP-7表达改变的影响。结果 20 mg/ml HSA刺激HKC细胞48 h后,HKC出现EMT样改变:CK表达下降(P<0.01)、Vimentin表达增高(P<0.01);HSA超载同时使HKC细胞内HOXA13、BMP-7表达下降。基因转染上调HKC细胞内HOXA13表达后,能够显著抑制HSA超载诱导的EMT发生(P<0.05)和BMP-7表达下降(P<0.05)。结论 HOXA13拮抗白蛋白超载诱导肾小管上皮细胞发生EMT的作用可能与HOXA13上调BMP-7表达有关。
Objective To investigate the effect of HOXA13 on HSA-induced mesenchymal transition (EMT) and bone morphogenetic protein-7 (BMP-7) expression in human renal tubular epithelial cells (HKC). Methods Human renal tubular epithelial cells (HKC) were cultured in vitro with 20 mg / ml human serum albumin for 48 h. Real-time PCR and Western blotting were used to detect the expression of cytokeratin (CK), vimentin Vimentin) and HOXA13 mRNA and protein expression. The expression of HOXA13 in human renal tubular epithelial cells was up-regulated by lipofection and the effect of HOXA13 expression on the changes of CK, Vimentin and BMP-7 induced by human serum albumin overload was observed. Results EMT-like changes were observed in HKC cells induced by 20 mg / ml HSA for 48 h: the expression of CK decreased (P <0.01) and the expression of Vimentin increased (P <0.01); the expression of HOXA13 and BMP-7 decline. Gene transfection could up-regulate the expression of HOXA13 in HKC cells significantly (P <0.05) and decrease the expression of BMP-7 induced by HSA overload (P <0.05). CONCLUSION: The effect of HOXA13 on the EMT induced by albumin overload in renal tubular epithelial cells may be related to the up-regulation of BMP-7 expression by HOXA13.