資源描述：
《游離亞油酸對(duì)胰腺腺泡細(xì)胞的影響及其機(jī)制的探討.doc》由會(huì)員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在工程資料-天天文庫(kù)。
1、游離亞油酸對(duì)胰腺腺泡細(xì)胞的影響及其機(jī)制的探討作者：王亞軍孫家邦李非孫海晨【摘要】目的探討游離亞油酸對(duì)胰腺腺泡細(xì)胞的影響及其可能的機(jī)制。方法分離胰腺腺泡細(xì)胞，分別與0?1、0.5、l.Onunol/L亞油酸共同孵育30、60、90min,測(cè)定細(xì)胞的LDH釋放率，DNA片段梯度分析腺泡細(xì)胞凋亡，流式細(xì)胞儀測(cè)定細(xì)胞凋亡率，Westernblot檢測(cè)腺泡細(xì)胞PKC的膜轉(zhuǎn)位，激光共聚焦測(cè)定細(xì)胞內(nèi)鈣的變化。結(jié)果腺泡細(xì)胞與亞油酸共同孵育后，臺(tái)盼藍(lán)染色顯示存在細(xì)胞損傷，LDH釋放率與亞油酸呈時(shí)間和劑量依賴(lài)關(guān)系。細(xì)胞總DNA凝膠電泳結(jié)果顯示1.0mmol/L亞油酸可出現(xiàn)DNA梯狀條帶，證明存在
2、細(xì)胞凋亡。同時(shí)，腺泡細(xì)胞凋亡率及PKC的膜轉(zhuǎn)位率，均與亞油酸呈劑量和時(shí)間依賴(lài)關(guān)系。另外，鈣離子是參與細(xì)胞損傷和PKC激活的因子之一，采用激光共聚焦方法檢測(cè)腺泡細(xì)胞內(nèi)鈣濃度，發(fā)現(xiàn)其升高與亞油酸呈濃度依賴(lài)性。結(jié)論亞油酸對(duì)胰腺腺泡細(xì)胞損傷存在劑量和時(shí)間依賴(lài)關(guān)系。游離亞油酸刺激腺泡細(xì)胞內(nèi)鈣升高和活化PKC誘導(dǎo)細(xì)胞凋亡可能是高脂血癥性胰腺炎的發(fā)病機(jī)制之一?！娟P(guān)鍵詞】胰腺腺泡細(xì)胞游離亞油酸細(xì)胞凋亡蛋白激酶C細(xì)胞內(nèi)鈣離子Explorationonfreefattyacidaffectingpancreaticacinarcel1anditspossiblemechanismWANGYaju
3、n,SUNJiabang,LIFei,SUNHaichen?DepartmentofGeneralSurgery,XuanwuHospitaiofCapitaiUniversityofMedicalScience,Beijingl00053,China【Abstract】ObjectiveToinvestigatetheeffectsoffreefattyacidonpancfeaticacinarcel1andexplorepossiblemechanism?MethodsPancreaticacinarcellsfromadultratswereincubatedwith
4、linoleicacidatconcentrationsof0?lmmol/L,0?5mmol/Land1?0nimol/Lfor30,60and90minutesrespectivelytomeasuretheLDHreleaserateofacinarcells?DNAladderana1ysiswasusedtoanalyzeapoptosisofacinarcelIs,flowcytometrytodetermineapoptosisrate,WesternblottodetectPKCmembranetranslocationandlaserscanningconf
5、ocalmicroscopy(LSCM)toobserveintracellularealciumchanges?ResultsAfterincubationwithlinoleicacid,pa.ncreaticacinarcellsstainedwithtrypanblueweredama.ged,withLDHreleaserateattimeanddosedependentfashionwithl1noleicacid?DNA1adderwasformedbyagarosegelelectrophoresiswhenacinarcellswereincubatedwi
6、th1.Ommol/Llinoleicacid,asprovedapoptosisofacinarcel1s.Also,acinarcel1apoptosisrateandPKCmembranetranslocationlevelwererelatedwithlinoleicacidatatimeanddosedependentfashion.Inaddition,intracellularcalciumwasoneofthefactorsleadingtocel1injurysnd卩KCactivation.LSCMshowedthattherisingofintracel
7、1ularealciumexistedatadosedependentfashionwithlinoleicacid?ConclusionsLinoleicaciddamagingacinarcel1isdependentlyrelatedtodoseandtime?Freefattyacidstimulatingincreaseofintracellularcalciuina.nda.ctivationofPKCinducingapoptosisisoneofpathogeneticmechanism
