CAJ | 학술논문

目的探讨地塞米松(Dex)及latrunculin A(Lat A)处理后人眼小梁细胞蛋白质表达的变化.方法正常供体人眼小梁细胞培养后用Dex及Lat A处理,经二维凝胶电泳分离小梁细胞蛋白质,分析凝胶电泳图谱,选取部分差异凝胶斑点并采用基质辅助激光解析电离飞行时间质谱(MALDI TOF MS)鉴定蛋白质.结果采用24cm IPG胶条获得正常人眼小梁细胞对照组、Dex组、Dex+Lat A组以及Lat A组的二维电泳凝胶图谱,得出不同培养条件下人眼小梁细胞蛋白质表达的差异.应用GDPi MALDI TOF MS得出不同培养条件下差异表达的蛋白质斑点并成功鉴定出其中的24种蛋白质,主要包括细胞骨架相关的蛋白、热休克蛋白、氧化还原相关的蛋白和糖代谢相关的蛋白4类蛋白质.ADLH和Rab蛋白在Lat A组的人小梁细胞中表达增加,但在Dex组表达减少,而热休克蛋白27(HSP27)和人CRMP2呈现相反的结果 .结论成功建立Dex及Lat A诱导人眼小梁细胞蛋白质表达图谱.Dex及Lat A能诱导人眼小梁细胞的蛋白质表达变化,可能与青光眼致病的分子机制相关.
목적 탐토지새미송(Dex)급latrunculin A(Lat A)처리후인안소량세포단백질표체적변화.방법 정상공체인안소량세포배양후용Dex급Lat A처리,경이유응효전영분리소량세포단백질,분석응효전영도보,선취부분차이응효반점병채용기질보조격광해석전리비행시간질보(MALDI TOF MS)감정단백질.결과 채용24cm IPG효조획득정상인안소량세포대조조、Dex조、Dex+Lat A조이급Lat A조적이유전영응효도보,득출불동배양조건하인안소량세포단백질표체적차이.응용GDPi MALDI TOF MS득출불동배양조건하차이표체적단백질반점병성공감정출기중적24충단백질,주요포괄세포골가상관적단백、열휴극단백、양화환원상관적단백화당대사상관적단백4류단백질.ADLH화Rab단백재Lat A조적인소량세포중표체증가,단재Dex조표체감소,이열휴극단백27(HSP27)화인CRMP2정현상반적결과 .결론 성공건립Dex급Lat A유도인안소량세포단백질표체도보.Dex급Lat A능유도인안소량세포적단백질표체변화,가능여청광안치병적분자궤제상관.
Background Researches have demonstrated that dexamethasone (Dex) can induce the changes of the function and structure of trabecular meshwork cells,and latrunculin A (Lat A) can enhance the outflow of aqueous humour and therefore low the intraocular pressure.Objective The aim of the present study is to investigate the effects of Dex and Lat A on the expression of protein in human trabecular meshwork cells.Methods Human trabecular meshwork cells were primarily cultured in DMEM using expand culture method and the fifth generation of cells were used to this experiment.Dex and/or Lat A were added in medium as 10~(-6)mol/L Dex group(Dex treating for 24 hours),Dex+Lat A group(10~(-6)mol/L Dex+2mmol/L Lat A for 24 hours),Lat A group(2mmol/L Lat A for 24 hours) and DMEM culture group.Two dimensional gel electrophoresis(2 DE) was used to compare the protein expressions among these four groups.Subsequently protein spots with different intensity were selected for mass spectrometry analysis.Results Four gel patterns of two dimensional gel electrophoresis of human trabecular meshwork cells from Dex,Dex+Lat A,Lat A and control groups were obtained.A good isolated result for majority of proteins in human trabecular meshwork cells was found in all of the four groups.An obvious expression difference of proteins in human trabecular meshwork cells was seen among the different culture conditions.Twenty four kinds proteins were identified by GDPiMALDI TOF MS,including cytoskeleton related proteins,heat shock proteins,redox related proteins,and proteins participating in carbohydrate metabolism.The expressions of aldehyde dehydrogenase(ADLH)and Rab were increased in Lat A group and decreased in Dex group,but HSP27 and hCRMP2 showed the contrary outcome.Conclusion This study construct the pattern of protein expression in human trabecular meshwork cells by using 2 DE.Dex and Lat A impact the protein expressions in human trabecular meshwork cells.