脱氧胆酸盐诱导培养的正常人食管粘膜上皮细胞凋亡。

Ru Zhang, Jun Gong, Hui Wang, Li Wang, Li-wei Ran
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引用次数: 0

摘要

目的:研究脱氧胆酸盐在体外诱导人正常食管粘膜上皮细胞凋亡中的作用,并探讨其分子机制。方法:用脱氧胆酸盐处理培养的正常人食管粘膜上皮细胞,采用TUNEL、DNA阶梯、PI-染色流式细胞术、Annexin V-FITC - PI染色和Western blotting检测细胞凋亡情况。结果:流式细胞术、TUNEL和DNA阶梯实验显示脱氧胆酸盐能诱导正常人食管粘膜上皮细胞凋亡,且呈剂量和时间依赖性。500微mol/L脱氧胆酸处理30 min后,流式细胞术检测到21.3%的细胞群显示caspase-3活性,显著高于对照(1.5%)。结论:脱氧胆酸盐可诱导培养的人食管粘膜上皮细胞凋亡。脱氧胆碱诱导的细胞凋亡与Aaspase-3激活、Bcl-2蛋白下调和Bax蛋白上调有关,与Fas- l /Fas无关。
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[Deoxycholate induces apoptosis in cultured normal human esophageal mucosal epithelial cells].

Objective: To study the effect of deoxycholate in inducing apoptosis of human normal esophageal mucosal epithelial cells in vitro and investigate the molecular mechanism.

Methods: Cultured normal human esophageal mucosal epithelial cells were treated with deoxycholate, and the cell apoptosis were evaluated with TUNEL, DNA ladder, flow cytometry with PI-staining, Annexin V-FITC conjugated with PI staining, and Western blotting.

Results: Flow cytometry, TUNEL and DNA ladder demonstrated that deoxycholate could induce apoptosis in normal human esophageal mucosal epithelial cells in a dose- and time-dependent manner. A percentage of 21.3% of the cell population treated with deoxycholate at 500 micromol/L for 30 min exhibited detectable caspase-3 activity shown by flow cytometry, which was significantly higher than the control level (1.5%, P<0.01). Western blotting suggested that deoxycholate down-regulated Bcl-2 protein expression and up-regulated Bax expression, but Fas was negative in the cells before and after deoxycholate treatment.

Conclusions: Deoxycholate could induce apoptosis in cultured human esophageal mucosal epithelial cells. Aaspase-3 activation, Bcl-2 protein down-regulation and Bax up-regulation are involved in deoxycholate-induced apoptosis, which does not involve Fas-L/Fas.

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