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目的:研究咖啡因对肝星形细胞活化状态和分泌功能的影响及其潜在机制。方法:选取人肝星形细胞LX-2进行体外实验。经浓度为0、5、10、20、30、40 m M的咖啡因处理后,通过Western印迹法分别检测不同浓度的咖啡因对肝星形细胞中α-平滑肌肌动蛋白和凋亡蛋白cleaved-PARP表达量的影响;分别用CCK-8实验和流式细胞仪检测咖啡因对肝星形细胞活性和凋亡的影响。结果:在咖啡因作用下,肝星形细胞表达α平滑肌肌动蛋白的量与对照组相比显著减少,表示其分泌功能受到明显抑制。CCK-8实验表明咖啡因可抑制肝星形细胞的活性,并呈现时间和浓度依赖性。流式细胞技术显示咖啡因作用浓度增高,凋亡细胞的比例随之增高,同时,cleaved-PARP的表达量与对照组相比明显增多,提示咖啡因可诱导肝星形细胞凋亡。结论:咖啡因可通过诱导凋亡降低肝星形细胞的活性,减少肝星形细胞中α平滑肌肌动蛋白的表达量,从而抑制肝星形细胞分泌胶原纤维的功能。
Objective: To investigate the effect of caffeine on activation status and secretion of hepatic stellate cells and its potential mechanism. Methods: Human liver astrocyte LX-2 was selected for in vitro experiments. After treated with caffeine at concentrations of 0, 5, 10, 20, 30 and 40 m M, Western blotting was used to detect the effects of different concentrations of caffeine on α-smooth muscle actin and apoptosis protein cleaved in hepatic stellate cells -PARP expression. The effects of caffeine on hepatic stellate cell activity and apoptosis were detected by CCK-8 assay and flow cytometry respectively. Results: Under the action of caffeine, the amount of α-smooth muscle actin expression in hepatic stellate cells was significantly decreased compared with the control group, indicating that the secretory function was significantly inhibited. CCK-8 experiments show that caffeine can inhibit hepatic stellate cell activity, and showed a time-and concentration-dependent. Flow cytometry showed that the concentration of caffeine increased, the proportion of apoptotic cells increased. At the same time, the expression of cleaved-PARP was significantly increased compared with the control group, suggesting that caffeine can induce hepatic stellate cell apoptosis. CONCLUSION: Caffeine can decrease the activity of hepatic stellate cells by inducing apoptosis, decrease the expression of α-SMA in hepatic stellate cells, and inhibit the secretion of collagen by hepatic stellate cells.