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《白藜蘆醇上調(diào)人肺泡上皮細胞SIRT1表達抑制高氧誘導(dǎo)的細胞凋亡.pdf》由會員上傳分享,免費在線閱讀,更多相關(guān)內(nèi)容在行業(yè)資料-天天文庫。
1、590細胞與分子免疫學(xué)雜志(ChinJCellMolImmuno1)2015,31(5·論著·文章編號:1007—8738(2015)05—0590—06白藜蘆醇上調(diào)人肺泡上皮細胞SIRT1表達抑制高氧誘導(dǎo)的細胞凋亡張春艷,李清平,康蘭,雷小平,翟雪松,趙帥,張嬋,董文斌(瀘州醫(yī)學(xué)院附屬醫(yī)院新生兒科,四川瀘州646000)[摘要]目的探討去乙酰化酶SIRT1激動劑白黎蘆醇(Res)對高氧誘導(dǎo)人肺泡上皮細胞(HPAEC)凋亡的保護作用。方法體外培養(yǎng)HPAEC,隨機分為對照組、高氧組、Res組。培養(yǎng)24h
2、,采用免疫細胞化學(xué)sP法檢測caspase-9、X聯(lián)鎖凋亡抑制蛋白(XIAP)、SIRT1蛋白的表達,采用MitoS0、JC一1標(biāo)記結(jié)合激光共聚焦顯微鏡分別檢測HPAEC線粒體內(nèi)活性氧(ROS)及其膜電位的變化,Westernblot法檢測SIRT1蛋白的表達,annexinV-FITC/碘化丙啶(PI)雙標(biāo)記結(jié)合流式細胞術(shù)檢測HPAEC細胞凋亡率的變化。結(jié)果對照組相比,高氧組HPAEC中caspase-9蛋白表達增強、線粒體內(nèi)ROS增加、細胞凋亡率增加,而XIAP、SIRT1蛋白的表達減少、膜電位降
3、低;與高氧組相比,Res組HPAEC中caspase-9的表達減弱、線粒體內(nèi)ROS產(chǎn)生減少、細胞凋亡率降低,XIAP、SIRT1蛋白表達增強、膜電位增高。結(jié)論Res通過上調(diào)HPAEC中SIRT1的表達,減少ROS的產(chǎn)生從而維持細胞膜電位抑制肺泡上皮細胞凋亡,減輕高氧所致肺損傷。[關(guān)鍵詞]高氧;肺泡上皮細胞;去乙酰化酶1;白藜蘆醇;凋亡[中圖分類號]R285,R392-33,Q255[文獻標(biāo)志碼】AResveratrolinhibitshyperxia·inducedcellapoptosisthrou
4、ghup-regulatingSIRT1expressioninHPAECsZHANGChunyan,LIQingping,KANGLan,LEIXiaoping,ZHAIXuesong,ZHAOShuai,ZHANGChan,DONGWenbinDepartmentofNeonatology,AfiliatedHospital,LuzhouMedicalCollege,Luzhou646000,China【AbstractJObjectiveToinvestigatetheprotectionefe
5、ctofsirtuin1(SIRT1)agonistresveratrol(Res)againsttheapoptosisofhumanpulmonaryalveolarepithelialcells(HPAECs)inducedbyhyperxia.MethodsTheHPAECs/nvitrowererandomlydividedintothreegroups:controlgroup,hyperxiagroup,Resgroup.After24-hourculture,theexpression
6、sofcaspase-9,X-linkedinhibitorofapoptosisprotein(XIAP),SIRT1proteinsweremeasuredbySPimmunohistochemistry.Thechangesofreactiveoxygenspecies(ROS)markedwithMitesOXTandmembranepotentialmarkedwithJC-1.nmitochondrionweredetectedbylaserscanningconfocalmicrosco
7、py.TheexpressionofSIRT1proteinwasdeterminedbyWesternblowing,andthechangeofcellapoptosisratewasanalyzedbyflowcytometrycombinedwithannexinV—FITC/PIstaining.ResultsComparedwiththecontrolgroup,theexpressionofcaspase-9,thegenerationofROSinmitochondrionofHPAE
8、Csandthecellapoptosisrateincreasedobviously.TheexpressionsofbothXIAPandSIRT1andmembranepotentialdecreasedevidentlyinthehyperxiagroup.Comparedwiththehyperxiagroup,theexpressionofcaspase-9,thegenerationofReSinmitochondrionofHPAECsa