磷酸化akt和adrp在高糖刺激的人肾小管上皮细胞中的表达的的研究

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1、中文摘要著差异，定量分析分别为0．99士O．04和0．99士0．03；激活状态的Akt，即磷酸化Akt在两组间的表达不同，高糖组明显高于正常糖组，分别为0．36圭O．05和0．99士0．07，差异有统计学意义。LY294002是P13K／Akt通路的特异性抑制剂，有效地减少了HKC细胞磷酸化Akt的表达，而对总Akt未见明显影响。3．高糖诱导人肾小管上皮细胞脂质合成增多油红O染色显示高糖刺激的HKC细胞胞浆内出现红染的细小颗粒，而正常糖组细胞内未见着色颗粒。经LY294002处理后脂滴形成明显减少。结论：1．高糖可诱导人肾小管上皮细胞脂

2、滴标记物ADRP的明显表达，提示糖尿病环境下，高糖是引起肾小管上皮细胞脂质代谢异常的重要因素。2．高糖可同时上调磷酸化Akt和ADRP表达，而抑制P13K／Akt通路可有效减少ADRP表达和细胞内脂滴，提示P13K／Akt通路可能参与了高糖刺激下的人肾小管上皮细胞的脂质代谢。谢关键词：糖尿病；肾小管上皮细胞；ADRP；P13K／Akt通路；脂质代研究论文Theexpressionofphospho-AktandADRPinhighglucose-stimulatedHKCcellsobjective：Diabeticnephropath

3、yisaseverecomplicationofdiabetesmellitusandmanyfactorsareinvolvedinthepathogenesisofthisdiseasesuchashyperglycemia,inflammatorycytokines，oxidativestressandSOon．Hi曲glucoseisanunderlyingfactorthataffectsnormalcells’physicalmetabolismandfunctionindiabetespatients．Someprevio

4、usstudieshaverevealedthathi曲glucosecanincreaselipiddropletsandextracellularmatrixaccumulationinrenaltubularcells．However,thedefinitemolecularmechanismsinvolvedintheprocesshavenotbeenfullyelucidated．ADRPisakindofsensitivemarkeroflipiddropletandcallbeusedtodetectearlyandsm

5、alldroplet．P13K／Aktpathwayisakindofmultiple—functionpathwayinvolvedincellproliferation,apoptosisandmetabolism．功eobjectiveofthisstudyistoinvestigatetheexpressionofphospho-AktandADRPinhi【曲glucose—stimulatedHKCcellsandexploretherelationshipbetweenP13K／Aktpathwayandlipidaccu

6、mulationinrenaltubularcells．Methods：1．HKCwaskindlyprovidedbyProfessorChenXiang-mei，DivisionofNephropathy,301Hospital，Beijing，China．CellsculturedinDMEMmediumcontaining30mmol／Lglucoseand1％fetalbovineserunl(FBS)wererandomlydividedintotwogroups：normalglucosegroup(glucose5ret

7、ool／L)，highglucosegroup(glucose30mmoFL)andLY294002group．．2．ImmunocytochemistryWasusedtodetecttheexpressionofADRPproteininhighglucose-treatedHKCcells．3．．nleexpressionofphospho-AktandtotalAktwasinvestigatedbythemethodofWesternblot．4．111eOilRed0stainingWasusedtodeterminethe

8、lipiddropletinhi．曲glucose—stimulated娅(Ccells．Results：一一—————————————————————————————————————————-—————一