Objective;To transfect hypoxia inducible factor-la (HIF-la) to human adipose-derived stem cells (hADSCs) and then coincubated with HK-2 cells induced by cisplatin. Methodology: The hADSCs were transfected by lentiviral vector containing EGFP-HIF-la and coincubated with HK-2 cells induced by cisplatin. The ultrastructural changes of HK-2 cells were observed by electron microscopy. Apoptosis index of HK-2 cells was tested by TUNEL. Western blot was performed to examine the expression of caspase-3 and Bcl-2 in HK-2 cells and also the expression of EPO, HO-1 and VEGF in hADSCs cells. The concentrations of NGAL, KIM-1, EPO, HO-1, VEGF and HIF-la in media were tested by ELISA. Results: The apoptosis in coincubation groups was decreased compared with the cisplatin group. The expression of caspase-3 in HK-2 cells in coincubation groups was lower than that in cisplatin group and the expression of Bcl-2 in HK-2 cells in coincubation groups was increased. The injury of HK-2 cells coincubated with hADSCs tranfected with HIF-la was lessened. The concentration of KIM-1 increased markedly in cisplatin-induced HK-2 group and that was decreased after coincubated with hADSCs. The expression of EPO, HO-1 and VEGF in hADSCs cells tranfected with H1F-la was increased compared with the vector transfected group. The concentration of HO-1 and VEGF in media of gene-transfected group was higher than that in vector transfected group and the variation of HIF-la and EPO were not obvious.Conclusion; The hADSCs could lessen the apoptosis of HK-2 cells induced by cisplatin, especially after transfected with HIF-la and the protection may be related to the higher expression of growth factors.%将低氧诱导因子1α(HIF-1α)基因质粒高效感染人脂肪源性干细胞(hADSCs),并与顺铂诱导的入近端肾小管上皮细胞(HK-2)共孵育,观察hADSCs对HK-2细胞损伤的影响.方法:以慢病毒为载体,将HIF-1α转染hADSCs,与顺铂诱导的HK-2细胞共孵育.电镜观察HK-2细胞形态、结构的变化；原位末端标记(TUNEL)检测细胞凋亡指数；Western blot检测HK-2细胞半胱氨酸天冬氨酸蛋白酶3(caspase-3)、B细胞淋巴瘤/白血病2基因(Bcl-2)的变化及hADSCs细胞促红细胞生成素(EPO)、血红素氧化酶1(HO-1)及血管内皮生长因子(VEGF)的表达,ELISA检测培养液中中性粒细胞明胶酶相关脂质运载蛋白(NGAL)和肾损伤分子1(KIM-1)的浓度,EPO、HO-1、VEGF和HIF-1α的浓度变化.结果:(1)与顺铂诱导组相比,共孵育后的HK-2细胞结构损伤减轻、凋亡细胞减少,caspase-3的表达低于顺铂诱导组、Bcl-2的表达水平升高；其中HIF-1α转染的hADSCs与HK-2细胞共孵育后,HK-2细胞损伤明显减轻,凋亡蛋白的变化更为明显.(2)顺铂诱导组培养液中NGAL、KIM-1浓度升高,以KIM-1变化明显(P＜0.05)；细胞共孵育后培养液中KIM-1浓度降低,转染组较明显.(3)转染后的hADSCs细胞内EPO、HO-1、VEGF的蛋白表达升高；培养液中HO-1、VEGF含量增高(P＜0.05),HIF-1α、EPO的含量变化不明显.结论:hADSCs与HK-2细胞共孵育后,可以减轻顺铂诱导的HK-2细胞结构损伤、抑制细胞凋亡,HIF-1 α转染hADSCs后其作用更为明显,可能与细胞保护因子的高表达有关.
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