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研究不同浓度的骨成形蛋白 7(BMP 7)对人近曲小管上皮细胞株HK 2凋亡的影响并初步探讨其可能的机制。应用Hoechst 332 5 8荧光染色、DNA电泳检测凋亡细胞的形态学和生化改变 ,流式细胞仪定量检测细胞的凋亡率 ,并观察凋亡特异性蛋白酶Caspase 3活性变化的情况。结果显示 ,HK 2细胞加入 1 0ng/mlTGF β处理 2 4h后 ,可明显抑制细胞生长并诱导细胞凋亡 ,呈现明显的凋亡形态学改变 ,胞核或核质内可见浓度致密的颗粒状荧光 ,DNA电泳呈现典型的“梯形条带” ,而预先加入 5 0ng/ml、 1 0 0ng/mlBMP 7共处理 2 4h后 ,可减轻TGF β对细胞的抑制作用 ,凋亡细胞亦减少 ,Caspase 3蛋白酶活性明显下降 ,细胞凋亡率下降。提示BMP 7对人肾小管上皮细胞凋亡的具有抑制作用 ,可能是通过抑制Caspase蛋白酶活性而实现的
To investigate the effect of different concentrations of bone morphogenetic protein 7 (BMP 7) on the apoptosis of human proximal tubule epithelial cell line HK 2 and its possible mechanism. The morphological and biochemical changes of apoptotic cells were detected by Hoechst 332 5 8 fluorescence staining and DNA electrophoresis. The apoptosis rate of the cells was quantified by flow cytometry and the changes of the activity of Caspase 3, an apoptosis-specific protease, were observed. The results showed that HK 2 cells treated with 10 ng / ml TGF β 2 for 24 h significantly inhibited cell growth and induced apoptosis, showing obvious apoptotic morphological changes. Density dense granular fluorescence was found in nuclei or nuclei , DNA electrophoresis showed a typical “trapezoidal band.” Pretreatment with 5 0ng / ml and 100ng / ml BMP 7 for 24 hours reduced the inhibitory effect of TGFβ on the cells and decreased the number of apoptotic cells. Caspase 3 Protease activity decreased significantly, the rate of apoptosis decreased. It is suggested that BMP7 can inhibit the apoptosis of human renal tubular epithelial cells possibly through the inhibition of Caspase protease activity