摘要
目的探讨牛磺酸熊脱氧胆酸(TUDCA)对高浓度葡萄糖诱导的人晶状体上皮细胞(hLECs)凋亡的作用及信号转导机制。方法hLECs在不同浓度葡萄糖培养液中培养24h,诱导建立hLECs凋亡模型,并采用不同浓度TUDCA(0.2、0.5、1.0、2.0mmol/L)进行干预。MTT法检测细胞增殖情况;Hoechst33258荧光染色法观察细胞凋亡形态学改变;AnnexinV-FITC/PI双染后流式细胞仪检测细胞凋亡率;Westernblot技术检测细胞葡萄糖调节蛋白78(GRP78)的表达。结果不同浓度葡萄糖培养细胞24h后,随着葡萄糖浓度的增加,细胞增殖抑制率亦升高(P<0.01)。高浓度葡萄糖(250mmol/L)培养24h可抑制hLECs的增殖活性,显著诱导hLECs凋亡(P<0.01),加入TUDCA共同培养后,hLECs凋亡率则显著降低(P<0.01),高浓度葡萄糖所引起的细胞GRP78蛋白表达也明显受到抑制(P<0.05)。结论内质网应激参与了高浓度葡萄糖诱导的hLECs凋亡,TUDCA可通过内质网应激途径抑制hLECs的凋亡,对hLECs产生保护作用。
Objective To investigate the effects and mechanisms of tauroursodeoxycholic acid(TUDCA)on human lens epithelial cells(hLECs)in a high glucose medium.Methods HLECs were incubated in DMEM with different concentrations of glucose for 24 h.A model of apoptotic hLECs was established and exposed to TUDCA(0.2,0.5,1.0,2.0 mmol/L)for 24 h.The proliferation of hLECs was determined by MTT.Morphologic evaluation of apoptotic cells was performed by Hoechst 33258 staining and the apoptosis rate was measured by flow cytometry(FCM)after staining of Annexin V-FITC and PI.Western blot was used to determine the expression of GRP78.Results After glucose treatment for 24 h,the inhibition of cell proliferation was increased with glucose concentration increasing(P〈0.01).Incubating the cells with high concentration of glucose(250 mmol/L)for 24 h inhibited the proliferation of hLECs and induced significant cells apoptosis(P〈0.01).The Apoptosis percentage of hLECs was decreased and expression of Bip/GRP78 was markedly inhibited after coincubation with TUDCA(P〈0.05).Conclusions Endoplasmic reticulum stress is involved in hLECs apoptosis induced by a high concentration of glucose.TUDCA inhibits hLECs apoptosis,thus protecting hLECs in vitro.The signal transduction mechanisms are related to endoplasmic reticulum stress.
出处
《山东大学学报(医学版)》
CAS
北大核心
2010年第3期74-77,82,共5页
Journal of Shandong University:Health Sciences