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目的:探讨胆管上皮细胞中Toll样受体3(TLR3)内源性活化对原发性胆汁性肝硬化损伤的影响。方法:体外培养人肝内胆管上皮细胞(HiBEC),并通过冻融处理制备坏死HiBEC样本。将死亡样本与活样本混合培养6h,以制备TLR3内源性活化样本,将其标记为观察组;以核酸酶处理坏死细胞及活细胞,并混合培养6h,以制备对照样本,将其标记为对照组。检测2组细胞凋亡率、TLR3及β干扰素Toll样受体结构域衔接蛋白(TRIF)表达水平、Caspase-3活性。结果:观察组凋亡率明显高于对照组,TLR3及TRIF相对表达量明显高于对照组,细胞Caspase-3平均表达量明显高于对照组,上述差异均有统计学意义(P<0.05)。结论:TLR3的内源性活化可导致HiBEC凋亡,在原发性胆汁性肝硬化的发生及发展中可能有一定促进作用。
Objective: To investigate the effect of endogenous activation of Toll-like receptor 3 (TLR3) on primary biliary cirrhosis in bile duct epithelial cells. Methods: Human hepatobiliary epithelial cells (HiBECs) were cultured in vitro and necrotic HiBEC samples were prepared by freezing and thawing. The dead samples were mixed with the live samples for 6h to prepare endogenous activation samples of TLR3, which were marked as observation group; necrotic cells and living cells were treated with nuclease and mixed for 6h to prepare control samples, which were marked as Control group. The apoptotic rate, TLR3 and Toll-like receptor domain adapter protein (TRIF) expression level and Caspase-3 activity of the two groups were detected. Results: The apoptosis rate in the observation group was significantly higher than that in the control group. The relative expression of TLR3 and TRIF in the observation group was significantly higher than that in the control group. The average expression of Caspase-3 in the observation group was significantly higher than that in the control group (P <0.05) . Conclusion: The endogenous activation of TLR3 can lead to the apoptosis of HiBEC and may promote the occurrence and development of primary biliary cirrhosis.