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生物工程学报j ournal s．i m ．ac．cncj b@ i m ．ac．cnChi n J Bi otech2008，Septem ber 25；24(9)：161 3-1 61 9Chi nese J ournal of Bi otechnol ogy@ 2008 Insti tute ofM i crobi ol ogy, CAS& CSM , Al lI SSN 1000· 3061ri ghts reservedLepti n对猪原代脂肪细胞脂解及其关键脂酶m RN A表达的影响李玉成，郑雪莉，杨公社西北农林科技大学动物科技学院动物脂肪沉积与肌肉发育实验室，杨凌712100摘要：I．xpti n是由脂肪组织分泌的内源因子，在调节机体能量平衡过程中起重要作用。Lepti n促进脂解的研究由来已久。但其作用机理尚不完善。本研究旨在通过系统检测关键脂酶m RN A的表达变化来探讨Lepfi n促进脂解的分子机理。运用形态学观察，油红O 染色和RT-PCR鉴定培养的猪原代脂肪细胞；甘油测定试剂盒和游离脂肪酸( FFA) 测定试剂盒分别检测甘油和FFA的释放；半定量RT-PCR检测关键脂酶m RN A的表达。结果显示：100 nm ol ／L的Lepti n可显著上调ATG L、TG H ．2、H SL．M G L和LPL m RN A的表达( P<0．01) ，但同时下调Peri l i pi n m RN A的表达( P<0．01) ；Lepdn呈浓度依赖性促进甘油的释放( P<0．01) ，但对FFA的释放影响不显著( 尸>O ．05) ．以上结果提示，Lepfi n可能主要通过上调ATGL、M GL、LPL和下调Peri l i pi n的表达促进猪原代脂肪细胞的脂解：同时推测，FFA释放的相对稳定可能是由Lepti n通过上调U cPs的表达而增加FFA的消耗引起的．关键词：Lepti n，脂解，猪，原代脂肪细胞，脂解关键脂酶Effects ofLepti non Porci nePri m ary Adi ocytes Li pol ysi sandm RN AExpressi onofKey Li pol yti c Enzym esYuchengLi ，Xuel i Zheng，and G ongshe YangLaboratory#Ani m al FatDeposi ti onandM uscl eDevel opm em , Col l egeofAni m al Sci encesandTechnol ogy, N orthw estA＆FU ni versi ty．Yangl i ng712100，Chi naAbstract：Lepti n，a cytoki ne predom i nantl ysecreted f romfatti ssue，pl aysal li m portantrol e i nregul ati ngorgani smel l 钉gybal ance．Lepti ncan sti m ul atel i pol ysi s，butthem echani smi s uncl ear．In order tostudythem ol ecul ar m echani smofl epti n sti m ul ati ng l i pol ysi s，w esystem i cal l ystudi ed the m RN Aexpressi onofkey l i pol yti c enzym es．M orphol ogi cal observati on，O i l Red Ostai ni ngandRT-PeR w ere used to i denti fy pi g pnrnary adi pocytes；com m erci alSem i quanfi tafi veRT- PCRw as used to detect thet uRN Aexpressi onofkey l i pol yti c enzym es．Theresul ts show ed thatl epti n up-regul atedthe m RN Aexpressi onof ATG L，TG H · 2，H SL，M G LandLPL( P<0．01) ，but dow n-regul atedthePeri l i pi nm RN Aexpressi on( P<O ．01) ．Atthe sam eti m e，l epti nprom otedthegl ycerol rel ease i n a dosedependent m am l er( P<0．01) ，buthadno effecton the FFArel ease(P>0．05)．Thesei ndi cate that l epti n m ay m ai nl y sti m ul atel i pol ysi si npi g pri m ary adi pocytes by up—regul ati ngtheexpressi onof ATG L，M G L，U ) L and dow n· regul ati ngtheexpressi onof Peri l i pi n．The unchangedFFArel easem aybe resul ted fromLepti nprom oti ngU CPsm RN Aexpressi onandi ncreasi ngFl FAexpendi ture．ki ts w ereused to m easure the gl yceroland FFA rel ease；100 nm oI／LRecei ved：J 如uary 30, 2008；ta：eol aod：Apri lSupported by：theN ati onal H i gh Technol ogy Research andD evel opm ent Program ( 863 Program ) of Chi na( N o．2006AAl 02138) ．Correspondi ngauthor：G ongsheYang．Tel ：+86—29· 87092430；Fax：+86—29· 87092430；E-m ai l ：gsyan9999@ hotm ai l ．com国家高技术研究发展计划( 863计划) 项目( N o．2006AAIO Zl 38) 资助．8，2008 万方数据