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目的:探讨牛磺熊脱氧胆酸(tauroursodeoxycholic acid,TUDCA)对棕榈酸(palmitate)诱导的INS-1细胞凋亡的保护作用。方法:分别用不同浓度棕榈酸(0.25、0.50、1.00 mmol/L)、棕榈酸(0.50 mmol/L)加TUDCA(100μmol/L)培养INS-1细胞24 h,用四甲基偶氮唑盐(MTT)法检测细胞活性,流式细胞术检测细胞早期凋亡,Western blot检测内质网应激相关蛋白GRP78的表达。结果:与对照组相比,棕榈酸组(浓度≥0.5 mmol/L)的INS-1细胞凋亡率显著上升(P<0.05);加TUDCA培养24 h以后,与棕榈酸组相比,INS-1细胞凋亡率显著下降(P<0.05)。此外,棕榈酸组(0.5 mmol/L)INS-1细胞内质网应激相关蛋白GRP78的表达显著上升(P<0.05);加TUDCA培养24 h以后,与棕榈酸组相比,INS-1细胞GRP78蛋白表达显著下降(P<0.05)。结论:TUDCA能够减少游离脂肪酸引起的INS-l细胞凋亡,对INS-1细胞发挥保护作用。而减少内质网应激蛋白的表达,缓解内质网应激可能是其作用机制之一。
Objective: To investigate the protective effect of tauroursodeoxycholic acid (TUDCA) on the apoptosis of INS-1 cells induced by palmitate. Methods: INS-1 cells were cultured with different concentrations of palmitic acid (0.25,0.50,1.00 mmol / L) and palmitic acid (0.50 mmol / L) plus TUDCA (100μmol / L) for 24 h. Cell viability was measured by MTT assay. Cell apoptosis was detected by flow cytometry. The expression of GRP78 in endoplasmic reticulum was detected by Western blot. Results: Compared with the control group, the apoptotic rate of INS-1 cells in palmitic acid group (≥0.5 mmol / L) was significantly increased (P <0.05). After adding TUDCA for 24 h, -1 cell apoptosis rate was significantly decreased (P <0.05). In addition, the expression of GRP78 in INS-1 cells treated with palmitic acid (0.5 mmol / L) increased significantly (P <0.05). After 24 h incubation with TUDCA, INS-1 GRP78 protein expression was significantly decreased (P <0.05). CONCLUSION: TUDCA can reduce INS-1 cell apoptosis induced by free fatty acids and exert a protective effect on INS-1 cells. Reducing the expression of ER stress protein and relieving endoplasmic reticulum stress may be one of its mechanisms.