摘要
目的：研究外源性同型半胱氨酸对体外培养的肾小球系膜细胞增殖、表型转化和产生细胞外基质的影响。方法：采用放射性同位素测定３ＨＴｄＲ掺入和细胞外信号调节激酶活性，流式细胞仪分析细胞周期，Ｗｅｓｔｅｒｎｂｌｏｔ方法进行α－平滑肌肌动蛋白检测，ＥＬＩＳＡ方法检测细胞上清液中细胞外基质含量。结果：不同浓度的同型半胱氨酸（０００５，００５，０５和１０ｍｍｏｌ／Ｌ）不同作用时间（８，１６和２４ｈ）对肾小球系膜细胞、细胞外信号调节激酶活性、α－平滑肌肌动蛋白表达和分泌型纤粘连蛋白、层粘连蛋白产生无明显作用。结论：同型半胱氨酸对体外培养的肾小球系膜细胞增殖、表型转化和细胞外基质产生无明显影响。
OBJECTIVE Tostudytheeffectofexogenoushomocysteine(HCY)onproliferation,phenotypechangeandextracellularmatrixproductionofculturedratglomerularmesangialcells(MC). METHODOLOGY RatmesangialcellswastreatedbyexogenousHCY.3HTdRincoporationandtheactivityofextracellularsignalregulatedkinase(ERK)wereassayedwithisotopicmethods,cellcycleanalysedbyflowcytometry.Theexpressionofsmoothmuscleactin(SMactin)wasdetectedbyWesternblot.Fibronectin(FN)andlaminin(LN)insupernatantofculturemediawereassayedbyinhibitionELISAmethods. RESULTS NoeffectonMC3HTdRincoporation(17333081849448vscontrol1709368,P>005)wasfoundwhenHCYincubatedwithMCatdifferentconcentrations(0005,005,05and10mmol/L)orofdifferenttimecourses(8,16and24hours).Theactivityofextracellularsignalregulatedkinase(ERK)werenotincreased(24647404137vscontrol362+91,P>005)Whencellsweretreatedwith05mmol/LHCYat2to60minutes.cellcyclesdetectedbyflowcytometrywerenotchangedbyHCYatdifferentconcentrations.SMactinexpressionwasnotenhancedbyHCYdetectedbyWesternblot.FNandLNinsuperna
出处
《肾脏病与透析肾移植杂志》
CAS
CSCD
1999年第3期228-231,共4页
Chinese Journal of Nephrology,Dialysis & Transplantation
关键词
同型半胱氨酸
系膜细胞
肾脏
肾小球疾病
homocysteinekidneymesangialcellstantofculturemediawerenotaffectedwhenmesangialcellsincubatedwithHCYattheconcentrationof05mmol/L. CONCLUSION HCYhasnoeffectsonproliferation,phenotypeandECMproductionofMC.
