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1、現(xiàn)代生物醫(yī)學進展www.shengwuyixue.comProgressinModernBiomedicineVoL14NO.8MAR.2014·1423·doi:10.13241/j.cnki.pmb.2014.08.006白藜蘆醇對氧糖剝奪/再灌注損傷的PC12細胞的保護作用及其作用機制冰劉宣王海嶸劉佳福陳向軍盧孔渺潘曙明(1上海交通大學醫(yī)學院附屬新華醫(yī)院急救中心上海200092;2復旦大學醫(yī)學院附屬華山醫(yī)院神經(jīng)內(nèi)科上海200040)摘要目的:探討白藜蘆醇對氧糖剝奪/再灌~(OGD/R)損傷的PC12細胞的保護作用及其機制。方法:體外培養(yǎng)PC12細胞,分為對照組。白藜蘆醇組,OGD/R組
2、及OGD+白藜蘆醇組。以改良的噻唑藍法測定細胞活性,采用AnnexinV—FITC/PI雙染法檢測細胞的凋亡率,用雙氫羅丹明(DHR)檢測細胞內(nèi)活性氧簇(ROS)的水平,采用蛋白印跡法(westernblot)分析SIRT1的蛋白表達情況。結(jié)果:與對照組相比,經(jīng)過OGD/R損傷后,細胞活力顯著降低。而在OGD/R的同時給予10tzmol/L的白藜蘆醇處理,可以明顯提高細胞活力。流式細胞儀檢測發(fā)現(xiàn),10t~mol/L的白藜蘆醇可以顯著地減少OGD/R引起的細胞凋亡,抑制細胞內(nèi)的ROS產(chǎn)生。westemblot的結(jié)果提示,與對照組比較,白藜蘆醇可提高SIRT1的蛋白表達水平。結(jié)論:白藜蘆醇可以通
3、過抑制ROS的產(chǎn)生和上調(diào)SIRT1的表達等機制而發(fā)揮其對抗氧糖剝奪/再灌注損傷的神經(jīng)保護性作用。關(guān)鍵詞:白藜蘆醇;氧糖剝奪/再灌注;細胞凋亡;ROS生成;SIRT1中圖分類號:R743。Q593.2文獻標識碼:A文章編號:1673—6273(2014)08—1423—05ProtectiveEffectofResveratrolontheInjuryofPC12CellsInducedbyOGD/RandItsMechanisms*L1-UXuan1,WANGHai-rongI,LIUJia-fuI,CHENXiang-jun2,LUKong-miaoIPANShu-mingJ(1Depar
4、tmentofEmergency,XinhuaHospital,ShanghaiJiaotongUnivemi~SchoolofMedicine,Shanghai,200092,China;2DepartmentofNeurology,HuashanHospital,FudanUniversitySchoolofMedicine,Shangha~20004o,China)ABSTRACTObjective:Toinvestigatetheprotectiveefectofresveratrol(RSV)ontheinjuryofPC12cellsinducedbyoxygenglucosed
5、eprivation/reperfusion(OGD/R)anditsmechanisms.Methods:PC12cellsculturedinvitroweredividedintocontrolgroup,RSVgroup,OGD/RgroupandOGD/R+RSVgroup.ThecellviabilitywasassessedbythemodifiedMTTmethod.AnnexinV—FITC/PIdoublestainingwasusedtodetectapoptosisrate.DHR123wasusedtomeasurecellularROSlevels.Western
6、blotwasusedtostudytheexpressionlevelofSIRT1.Results:Comparedtothenormalcon~olgroup,thecellviabilitydecreasedsignificantlyafterOGD/Randgiving10p,mol/LRSVduringOGD/Rcouldsignificantlyincreasecellviability.Basedontheresultsofflowcytometry,wefound10ixmol/LRSVmarkedlyinhibitedOGD/R.inducedapoptosisandre
7、ducedintracellularROSgeneration.ResultofWestemblotshowedtllatRSVcouldincreasetheexpressionofSIRT1comparedwithcontrolgroup.Conclusion:RSVwasfotmdtohaveaneuroprotectiveefectagainstOGD/R-inducedinjurybyattenua