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《高糖环境诱导胰岛表达p25引起胰岛细胞损伤的研究.pdf》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库。
1、·208·宁夏医学杂志2015年3月第37卷第3期NingxiaMedJ,Mar.2015,Vol37,No.3Doi:10.13621/j.1001—5949.2015.03.0208·论著·高糖环境诱导胰岛表达p25引起胰岛细胞损伤的研究罗红艳,王岩。,刘冉冉,张彬,毕逢辰,张霞,李博,郑亚莉[摘要]目的探讨在高糖环境中p25的表达及其对胰岛细胞损伤及胰岛素分泌的影响。方法实验分为db/db小鼠组(糖尿病组,n:5)和C57/BL小鼠组(正常对照组,n=5),进一步将从C57/BL小鼠胰岛分离的胰岛细胞组分为正常糖浓度糖刺组(5mM)和高浓度糖刺激组(30raM)。研究对象均为
2、雄性小鼠,鼠龄l3周,进行胰岛(islets)分离,培养及胰腺组织病理切片。用免疫印迹测定蛋白表达,酶联免疫法测定胰岛素分泌水平,免疫组化染色测定胰岛细胞凋亡。结果p25在C57/BL组和C57/BL正常糖浓度刺激组的胰岛细胞均无表达,与之比较,p25在db/db组和C57/BL高糖刺激组的胰岛细胞均有明显的表达(P<0.05);与C57/BL组相比较,cleavedeaspase3在db/db组胰岛细胞中的表达明显增加(P<0.05);与C57/BL组和C57/BL正常糖浓度组相比较,胰岛素分泌在db/db组和C57/BL高糖刺激组均明显减少(P<0.05)。结论高糖环境可刺激胰
3、岛表达p25,引起胰岛细胞凋亡,减少胰岛素分泌。[关键词】p25;糖尿病模型;胰岛;胰岛素;细胞凋亡[中图分类号]R692[文献标识码]AHighglucoseinducesp25expressionandresultstheisletcellsinjuryindiabeticmiceLUOHongyan,WANGYah,Ranran,ZHANGBin,BIFengchen。,ZHANGXia,12Bo,ZHENGYali.1.DepartmentofNephrology,NingxiaPeople’sHospital,Yinchuan750021,China;2.NingxiaM
4、edicalUniversity,Yinchuan750004,ChinaCorrespondingauthor:ZHENGYali,Email:yalinew@yahoo.com[Abstract]ObjectiveTodeterminetheexpressionandeffectsofp25inisletcellsanddiabeticmicemode1.MethodsUsingmale,age13weeksdb/dbmiceasdiabetesmodel(db/dbgroup(n=5)andthesameage,maleC57/BLmiceascontrolgroup(n=5
5、).Furthermore,subgroupedthecontrolgroupintohighglucose(30mM)andnormalglucose(5mM).Afterfeedoneweek,theisletsweredissectedandpurified,thencultured.ThepancreaseswerefixedforthepathologicalexaminationandthelevelsofinsulinsecretionweredetectedwithELISA,andapoptosisofisletcellswereanalyzedbyimmunoc
6、hemistrystaining.ResultsIndb/dbgroupandhighglucosetreatedgroup(30mM),theexpressionofp25wasdetectedinisletculturecells,whiletherewerenoexpressionofp25inthecontrolgroupandlowglucose(5mM)group.Comparingtothecontrolgroup,theexpressionofcleavedcaspase3whichwasamarkerofcellap—optosis,wasincreasedobv
7、iouslyinisletculturecellsinthedb/dbgroup(P<0.05).Insulinsecretionsignificantlydecreasedinthedb/dbgroupandhighglucosetreatedgroup(30mM)(P<0.05).ConclusionHighglucosestimulationinducestheexpressionofp25inisletscells,causesisletcellsapopto
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