温度对体外大鼠睾丸支持细胞增殖活力及occludin蛋白表达影响

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1、AbstractHightemperaturereducestheproliferationandoccludinexpressioninratSertoilcellsinvitroObjectiveThepurposeistofurtherexploretheactionmechanismofvarIoustemperaturesonmaleinfertilityorsubfertilityviaa11observationontheeffectofdifferenttemperaturesontheproliferationandocclu

2、ding(OCLN)expressionofinvitroratsertolicells(SOs)．MethodsFirstlv，isolateSCsfromthetesticleofmaleWistarratandtestitbyoilredOsmimngandimmunohistochemistryFasL．Secondly，setupSCsintoonecontrolgroup(34℃)andfiveexperimentalgroups(35℃、36℃、37℃、38℃、39℃)．CultivateSCsundereachtemperatu

3、reforfourdays．Thirdly，tomakeacomparison，applyCCK-8assaytomeasurecellproliferation；observecellmorphologyandstIucnlreafterHematoxylin．Eosinstaimng，underinvertedmicroscopeandlightH1icroscope：thendetermineOCLNexpressionlevelthroughwesternblottingandcellimmunofluorescence．Results

4、AsaresultofoilredOandFasLtest，thepurityofisolatedSCswas96．20士1．95％inthisexperiment．CCK．8assayindicatedthatproliferationofSCsincreaSedasthetemperaturewasrisingbetweentherangeof34℃and36℃，anddecreaSedasthetemperatureWasrisingintherangeof36～39℃．Meanwhile，pyknoficnucleiandfragmen

5、tationofSCsweremoreobviousunderatemperatureabove36℃．Otherwise，westernblottingandimmunofluorescenceshowedapeakofOCLNexpressionlevelunder36℃．Asacontrast，OCLNexpressionleveldecreasedwhilethetemperaturewasgoingupover36℃fP<0．01)．ConelusionsTheresultofthisstudyindicatedthatahigher

6、temperature(37℃，38℃or39℃)inhibitedtheproliferationofratSCsinvitro，anddecreasedtheexpressionofOCLN，fromwhich，wecarltentativelyspeculatethatallighIxtemperatureabove36。CmayreducemalefertilitybyaffectingtheproliferationofSCsandintegrityoftightjunctionbetweenSCsorSCsandothercells

7、．Postgraduatestudent：Hujianting(Urology)Directedby：Prof．Xin—shengWangKeyWords：temperature；Sertoliceil；occludin；blood-testisbarrie：r；spermatogenesis目录引言⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯．．1第1章材料和试剂⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯21．1试验动物⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯21．2主要实验试剂⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯

8、⋯⋯⋯⋯⋯⋯⋯21．3主要仪器设备与工具⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯3第2章实验方法⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯⋯52