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山东大学学报(医学版) ›› 2010, Vol. 48 ›› Issue (3): 74-77.

• 论文 • 上一篇    下一篇

牛磺酸熊脱氧胆酸对高糖环境下晶状体上皮细胞的保护作用

蒋欣桐1,蔡可丽1,郭媛媛1,王荣2   

  1. 山东大学齐鲁医院 1. 眼科; 2. 教育部和卫生部心血管重构与功能研究重点实验室, 济南 250012
  • 收稿日期:2009-12-17 出版日期:2010-03-16 发布日期:2010-03-16
  • 通讯作者: 蔡可丽(1961- ),女,教授,主要从事白内障及眼表疾病的研究
  • 作者简介:蒋欣桐(1983- ),女,硕士研究生,主要从事白内障及眼表疾病的研究。

Protective effects of tauroursodeoxycholic acid on human lens epithelial cells in a high glucose environment

JIANG Xin-tong 1,CAI Ke-li 1,GUO Yuan-yuan 1,WANG Rong 2   

  1. 1. Department of Ophthalmology; 2. Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Public Health, Qilu Hospital of Shandong University, Jinan 250012, China
  • Received:2009-12-17 Online:2010-03-16 Published:2010-03-16

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;Annexin VFITC/PI双染后流式细胞仪检测细胞凋亡率;Western blot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。

关键词: 白内障;牛磺酸熊脱氧胆酸;内质网应激;细胞凋亡;葡萄糖

Abstract:

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 24h. A model of apoptotic hLECs was established and exposed to TUDCA(0.2,0.5,1.0,2.0mmol/L)for 24h. 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 24h, the inhibition of cell proliferation was increased  with glucose concentration increasing(P<0.01). Incubating the cells with high concentration of glucose(250mmol/L)for 24h 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.

Key words: Cataract;Tauroursodeoxycholic Acid;Endoplasmic reticulum;Apoptosis;Glucose

中图分类号: 

  • R776.1
[1] 郭媛媛,蔡可丽,蒋欣桐. 褪黑素对抑制过氧化氢诱导大鼠白内障形成的实验研究[J]. 山东大学学报(医学版), 2010, 48(5): 67-71.
[2] . ReSTOR多焦点人工晶体的临床应用研究[J]. 山东大学学报(医学版), 2009, 47(7): 103-106.
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