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1、天然腦活素促細(xì)胞增殖及抗細(xì)胞衰老作用作者:李映紅吳正治吳偉康李明張曉麗陳茵楊敏【摘要】目的:探討天然腦活素對(duì)人胚肺二倍體成纖維細(xì)胞(MRC-5)增殖能力和復(fù)制性衰老的影響及其機(jī)理。方法:光鏡下觀察MRC-5細(xì)胞衰老表型,常規(guī)計(jì)數(shù)細(xì)胞代齡、傳代速度,MTT法檢測(cè)細(xì)胞增殖活度,X?Gal染色法觀察細(xì)胞衰老情況,計(jì)數(shù)衰老細(xì)胞百分率。結(jié)果:天然腦活素可維持細(xì)胞的非衰老表型,增加MRC-5細(xì)胞代齡,天然腦活素孵育MRC-5細(xì)胞的細(xì)胞增殖能力較空白老齡對(duì)照組明顯升高(P<0.05),細(xì)胞的增殖速度顯著加快。另外,天然腦活素連續(xù)
2、培養(yǎng)的細(xì)胞在老齡階段衰老細(xì)胞數(shù)顯著低于老齡對(duì)照細(xì)胞(P<0.01)0結(jié)論:天然腦活素可升高M(jìn)RC-5細(xì)胞增殖能力,有效延緩細(xì)胞的復(fù)制性衰老。【關(guān)鍵詞】天然腦活素;人胚肺成纖維細(xì)胞;增殖活度;復(fù)制性衰老;延緩(Abstract)ObjectiveToexploretheeffectsofNaturalCerebrolysinoncellproliferationactivityandreplicativesenesceneeofhumanembryoniclungdiploidfibroblasts(MRC-5cel
3、ls)anditsmechanism?MethodsToobservethechangesofsenescentphenotypebylightmicroscope,countthepassageagesandvelocity,detectthepotentialofcellproliferationbyMTT?ThechangesofcellssenesceneeofNaturalCerebrolysin?effectedMRC-5cellswerestudiedbyX-Galstainingmethod,and
4、thepositivecellratesofsenescencecellsweredetectedandanalyzed?ResultsNaturalCerebrolysinmaintainedunalteredcellmorphologyandincreasedlifespanofMRC-5byatleast18-21passages?Comparingwiththecontrolseniumcells,theproliferationpotentialofMRC-5cellsincubatedwithNatur
5、alCerebrolysinwasincreasedobviously(P<0.05),andthepassagevelocitywasenhanceddoubleofthatofthecontrolseniumcells(P<0.01).Inaddition,thepositivecellrateofsenesceneecellswassignificantlylowerinlatepassagecellsinsequeneedmultigenerationcultureinNaturalCerebr
6、olysinthanthoseofcontrolseniumcells(P<0.01),andsimilartothoseofyoungcells?ConelusionAsaresult,weconeludedthatNaturalCerebrolysincouldnotablypromotetheproliferationactivityofMRC-5cellsanddelaythecellsreplicativesenesceneeefficiently.(KeyWords〕NaturalCerebrol
7、ysin;Humanembryoniclungdiploidfibroblasts;Proliferationactivity;Delay;Replicativesenescence衰老(senescence)是細(xì)胞的重要生命現(xiàn)象之一,絕大多數(shù)正常細(xì)胞被認(rèn)為僅有有限的分裂能力,當(dāng)其經(jīng)過一定的倍增之后,將進(jìn)入終末增殖狀態(tài),稱為細(xì)胞衰老。一般來講,人類胚胎的成纖維細(xì)胞在達(dá)到衰老時(shí),能夠復(fù)制50代左右(PopulationDoublings,Pds),由于這種衰老是由分裂傳代而達(dá)到的,故又稱之為復(fù)制性衰老(replica
8、tivesenescence)[1]o探討細(xì)胞衰老的分子機(jī)理是人類認(rèn)識(shí)自我研究的重要組成部分,從分子水平研究衰老過程中基因及調(diào)控變化,已成為目前衰老機(jī)理研究的主要方向,同時(shí)研究開發(fā)延緩衰老的藥物將為提高人類生存質(zhì)量提供重要保證,也為衰老相關(guān)疾病的防治提供理論基礎(chǔ)和實(shí)驗(yàn)依據(jù)。天然腦活素為結(jié)合臨床經(jīng)驗(yàn)和現(xiàn)代藥效學(xué)正交實(shí)驗(yàn)優(yōu)化篩選而成,本研究在以往基礎(chǔ)上以人胚肺二