[1]李宝海,陈媛媛,周健. 内质网应激对糖尿病小鼠晶状体上皮细胞发生上皮间质转分化的作用[J].眼科新进展,2015,35(3):205-209.[doi:10.13389/j.cnki.rao.2015.0056]
　LI Bao-Hai,CHEN Yuan-Yuan,ZHOU Jian. Effects of endoplasmic reticulum stress on EMT of lens epithelial cells in diabetic mice[J].Recent Advances in Ophthalmology,2015,35(3):205-209.[doi:10.13389/j.cnki.rao.2015.0056]
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 内质网应激对糖尿病小鼠晶状体上皮细胞发生上皮间质转分化的作用
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
35卷
期数:
2015年3期
页码:
205-209
栏目:
实验研究
出版日期:
2015-03-05

文章信息/Info

Title:
 Effects of endoplasmic reticulum stress on EMT of lens epithelial cells in diabetic mice
作者:
 李宝海; 陈媛媛; 周健
 710032　陕西省西安市，第四军医大学西京医院眼科、全军眼科研究所
Author(s):
 LI Bao-Hai ;  CHEN Yuan-Yuan;  ZHOU Jian
 Department Ophthalmology,Xijing Hospital,the Fourth Military Medical University,Eye Institute of Chinese PLA,Xi’an 710032.Shaannxi Province．China
关键词:
 ; 上皮间质转分化; 内质网应激; 4-苯基丁酸; 白内障; 糖尿病
Keywords:
 ; epithelial-mesenchymal transition;  endoplasmic reticulum stress ;  4-Phenylbutyric acid ;  cataract ;  diabetes mellitus
DOI:
10.13389/j.cnki.rao.2015.0056
文献标志码:
A
摘要:
 目的　探讨内质网应激对糖尿病小鼠晶状体上皮细胞发生上皮间质转分化（ｅｐｉｔｈｅｌｉａｌ-ｍｅｓｅｎｃｈｙｍａｌｔｒａｎｓｉｔｉｏｎ，ＥＭＴ）的作用。方法　４２只雄性Ｃ５７ＢＬ／６Ｊ小鼠随机分为正常对照组、糖尿病模型组和糖尿病＋４-苯基丁酸（４-ＰＢＡ）干预模型组。糖尿病模型小鼠由腹腔注射ＳＴＺ建立，糖尿病＋４-ＰＢＡ干预模型组给予４-ＰＢＡ溶液灌胃。干预过程中监测各组小鼠体质量及随机血糖水平变化。干预开始后每４周散瞳后于裂隙灯显微镜下观察晶状体混浊情况并照相记录，第１２周摘取眼球于体式显微镜下观察晶状体混浊情况。在干预的第１、４、８和１２周，采用Ｗｅｓｔｅｒｎｂｌｏｔ方法检测晶状体上皮中葡萄糖调节蛋白７８（ｇｌｕｃｏｓｅｒｅｇ-ｕｌａｔｅｄｐｒｏｔｅｉｎ，ＧＲＰ７８）、α-平滑肌肌动蛋白（α-ｓｍｏｏｔｈｍｕｓｃｌｅａｃｔｉｎ，α-ＳＭＡ）及Ｅ-钙黏蛋白（Ｅ-ｃａｄｈｅｒｉｎ）表达水平。结果　正常对照组小鼠体质量增加明显，观察期结束时体质量为（３０．２１±１．４７）ｇ，糖尿病模型组及糖尿病＋４-ＰＢＡ干预模型组小鼠体质量增长较正常对照组小鼠缓慢，１２周末时分别为（２５．３２±０．７３）ｇ及（２５．０７±０．６７）ｇ。随机血糖监测结果显示糖尿病模型组小鼠各时间点血糖均高于正常对照组（ｔ＝１３．５２、１９．４５、１８．８６、２１．１８、２１．５６，均为Ｐ＜０．０５），糖尿病＋４-ＰＢＡ干预模型组血糖各时间点均高于正常对照组（ｔ＝１５．２３、１８．７８、１３．０４、１５．０６、１３．９０，均为Ｐ＜０．０５），第４周起低于糖尿病模型组（ｔ＝５．８１９、６．１２０、７.６６４，均为Ｐ＜０．０５）。裂隙灯显微镜下检查结果显示，正常对照组小鼠晶状体维持透明，糖尿病模型组小鼠第８周起晶状体出现浅层皮质点状混浊，糖尿病＋４-ＰＢＡ干预模型组小鼠８周时晶状体出现轻度混浊，但程度较糖尿病模型组轻。离体观察正常对照组晶状体保持透明，糖尿病模型组及糖尿病＋４-ＰＢＡ干预模型组晶状体均出现点状混浊，糖尿病＋４-ＰＢＡ干预模型组混浊程度较轻。Ｗｅｓｔｅｒｎｂｌｏｔ结果显示，与正常对照组相比，糖尿病模型组ＧＲＰ７８蛋白表达量在第１２周时明显增高（ｔ＝７．１５３，Ｐ＜０.０５）；８周及１２周时α-ＳＭＡ表达量明显升高（ｔ＝３．５５９、４．０８４，均为Ｐ＜０．０５），各时间点Ｅ-ｃａｄｈｅｒｉｎ表达量均明显降低（ｔ＝４.３５０、６．１３９、７．７７０、３．４８６，均为Ｐ＜０．０５）；糖尿病＋４-ＰＢＡ干预模型组ＧＲＰ７８蛋白表达量低于糖尿病模型组，且在第８周和１２周差异均有统计学意义（ｔ＝８．６００，１１．５３，均为Ｐ＜０．０５）；α-ＳＭＡ相对表达量在第１２周时明显下降（ｔ＝４．３５７，Ｐ＜０．０５），Ｅ-ｃａｄｈｅｒｉｎ相对表达量在增高且达到正常水平，各时间点差异均有统计学意义（ｔ＝４．３６０、７．６３３、８．４５０、５．８５３，均为Ｐ＜０．０５）。糖尿病＋４-ＰＢＡ干预模型组不同时间点比较，ＧＲＰ７８表达量在８周时达到最低。结论　利用分子伴侣４-ＰＢＡ抑制糖尿病小鼠内质网应激水平能够抑制糖尿病引起的晶状体上皮细胞发生ＥＭＴ。
Abstract:
 Objective To investigate the effects of endoplasmic reticulum stress on epithelial-mesenchymal transition of the lens epithelial cells in diabetic mice. Methods Forty-two male mice were randomly divided into normal group , diabetes mellitus (DM) group and DM +4-Phenylbutyric acid (4-PBA) group. Diabetic models were induced by streptozotocin ( STZ) . After the models were established.4-PBA were givenby means of gastric perfusion. Lens opacification was observed with slit-lamp microscope every 4 weeks. Photos of the lenses in vitro were taken with stereomicroscope at 12 weeks. By the end of I week,4 weeks , 8 weeks and 12 weeks , the expressions of glucose regulated protein 78 ( GRP78 ) .d一smooth muscle actin (d—SMA) and E-cadherin were detected by Westem blot assay. Results The body mass in normal group was obviously increased , which at the final follow-up was ( 30. 21 + I . 47 ) g , the body mass in DM group and 4-PBA group were lower than that in normal group , which were ( 25. 32 + 0. 73)g and (25. 07 + 0. 67) g. Blood glucose level in DM group presented to be much higher than those in normal group at each time point ( t = 13. 52 . 19. 45 . 18. 86 ,21. 18 . 21. 56 , all P < 0. 05 ) ,4-PBA group were higher than normal group ( t = 15. 23. 18. 78 . 13. 04 . 15. 06 , 13. 90 , all P < 0. 05 ) .while it showed relatively lower in 4-PBA group compared with DM group from 4 weeks ( t = 5. 819 .6. 120 .7. 664 . all P < 0. 05 ) . Slit-Iamp examination at 8 weeks and 12 weeks showed punctate opacification of lenses in DM and 4-PBA groups , while the lenses in normal group remained clear. At 8 weeks , the lenses in normal group remained clear, but the lenses in DM and 4-PBA groups appeared punctate opacification in vitro. Western blot assay deternuned that. compared with normal group , the relative expressions of GRP78 in DM group increased at 12 weeks ( t = 7. 153 . P < 0. 05 ) , the expression of e/-SMA increased at 8 weeks and 12 weeks ( t = 3. 559. 4. 084 . all P < 0. 05 ) . while the expressions of E-cadherin reduced ( t = 4. 350, 6. 139 . 7. 770 .3. 486 . all P < 0. 05 ) . In 4-PBA group , the expressions of GRP78 were lower than those in DM group, which were significantly different at 8 weeks and 12 weeks ( t = 8. 600 ,11. 53 , all P < 0. 05 ) . and the expression of d一SMA reduced at 12 weeks ( t = 4. 357 .P < 0. 05 ) ; The expression of E-cadherin was higher than that in DM group , which even reached normal level, the si~ificant differences were found at all time points ( t = 4. 360 .7. 633 , 8. 450 .5. 853 , all P < 0. 05 ) . As the extension of time , the expression of GRIy78 protein was presented downward trend . and reached bottom at 8 weeks after treated with 4-PBA. Conclusion Inhibition of endoplasmic reticulum stress by 4-PBA may suppress epithelial-mesenchymal transition in lens epithelial cells of diabetes mellitus mice.

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备注/Memo

备注/Memo:
国家自然科学基金资助（编号：８１３７０９９８）；陕西省科技统筹创新工程项目资助（编号：２０１２ＫＴＣＱ０３-０３）
更新日期/Last Update: 2015-03-04