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[摘要]
摘要:目的 研究雷公藤红素(Celastrol, CEL)对胆管细胞(Human biliary epithelial cells)的毒性作用及机制。方法 通过CCK-8实验及显微镜观察记录CEL对细胞形态和活力的影响;通过细胞划痕实验检测CEL对细胞迁移能力的影响;通过流式细胞术检测CEL对细胞细胞周期影响和对细胞凋亡的诱导作用;通过qRT-PCR和Western Blot检测CEL对凋亡相关基因Caspase 3、Caspase 8、Bax、Bcl2基因的表达情况。结果 CEL 在400-2000nM下,抑制细胞增殖,改变细胞形态;在200-800nM下,抑制细胞的迁移能力;在800-1200nM下,出现G0/G1期阻滞作用;在400-1200nM下,诱导细胞凋亡并增加相关基因的表达量。结论 雷公藤红素可通过影响胆管细胞活力,改变其迁移能力,阻滞细胞周期并促进细胞凋亡的方式产生胆管毒性。
[Key word]
[Abstract]
Abstract: Objective To investigate the toxicity and mechanisms of Celastrol (CEL) on human biliary epithelial cells. Methods The effects of CEL on cell morphology and cell viability changes were observed by CCK-8 experiment and microscope. Cell scratch experiment was used to detect the effect of CEL on cell migration. The effects of CEL on cell cycle and cell apoptosis were detected by flow cytometry. The Mrna and protein expression of CEL on apoptosis-related genes Caspase 3, Caspase 8, Bax and Bcl2 were detected by qRT-PCR and Western Blot. Results CEL inhibited cell proliferation and changed cell morphology at 400-2000nM. At 200-800nM, cell migration was inhibited. At 800-1200nM, G0/G1 phase was prevented. At 400-1200 nM, cell apoptosis was induced and the expression of apoptosis-related genes was increased. Conclusion CEL showed cholangiocyte toxicity through affecting cell viability, migration, preventing cell cycle and promoting cell apoptosis of human biliary epithelial cells.
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[基金项目]
北京市科技新星;北京市双一流课题