東南大學(xué)學(xué)報(bào)(醫(yī)學(xué)版)674J Southeast Univ( Med Sci Edi2012,Dec;31(6):674-680論著乙醇對肌肉衛(wèi)星細(xì)胞 myogenIn表達(dá)的影響沈干1,吳京2, Gordon huggins(1.南京醫(yī)科大學(xué)第二附院東院整復(fù)外科,江蘇南京21003;2. New England Medical Center,Tufts University, Massachusetts Boston 02115)摘要]目的:探討慢性酒精中毒過程中乙醇對肌肉衛(wèi)星細(xì)胞 myogen表達(dá)的影響及其在酒精性肌肉損傷中的作用。方法:(1)體外實(shí)驗(yàn):取 Sprag- Dowley大鼠,原代培養(yǎng)大鼠肌肉衛(wèi)星細(xì)胞,設(shè)實(shí)驗(yàn)組(乙醇作用組)與對照組(正常培養(yǎng)液對照組),培養(yǎng)細(xì)胞至80%融合時,更換為分化培養(yǎng)液,對實(shí)驗(yàn)組與對照組衛(wèi)星細(xì)胞形成的肌管計(jì)數(shù),并觀察衛(wèi)星細(xì)胞 myogenin的表達(dá)。(2)動物實(shí)驗(yàn):予小鼠乙醇飲食(Ⅰ個月)制造乙醇中毒動物模型,在損傷后肢肌肉后第27天取組織切片觀察損傷部位肌肉衛(wèi)星細(xì)胞增殖及新肌束形成情況。結(jié)果:實(shí)驗(yàn)組肌管計(jì)數(shù)及衛(wèi)星細(xì)胞 myogenIn的表達(dá)均低于對照組(P<0.05),但乙醇飲食小鼠損傷肌肉標(biāo)本中衛(wèi)星細(xì)胞及新肌束數(shù)量與正常飲食小鼠差異無統(tǒng)計(jì)學(xué)意義(P>0.05)。結(jié)論:乙醇可以通過降低肌肉衛(wèi)星細(xì)胞的 myogenin表達(dá)抑制其肌性分化,減少衛(wèi)星細(xì)胞形成肌管的數(shù)量。關(guān)鍵詞]肌肉;衛(wèi)星細(xì)胞;乙醇; myogenIn;分化中圖分類號]R685.2文獻(xiàn)標(biāo)識碼]A[文章編號]1671-6264(2012)06-0674-07doi:10.3969/j.issn.1671-6264.2012.06.003Effect of ethanol to myogenin expression in muscular satellite cellsSHEN Gan, WU Jing, gordon Huggin(1. Department of Plastic and Reconstructive Surgery, the Second Affiliated Hospital of Nanjing MedicalUniversity, Nanjing 210003, China; 2. New England Medical Center, Tufts University, BostonAbstract]Objective: To investigate the effect of ethanol to myogenin expression in muscular satellite cells and it'srole in alcoholic muscle atrophy. Methods: 1 )In vitro study, Sprage-Dowley rats were used, satellite cells wereharvested and cultured from rat muscle, experimental group( ethanol culture medium )and control group( normalculture medium )were set. When cells became 80% confluent, growth culture medium was changed into differentiationmedium. To evaluate the effect of ethanol, myotube quantitation and Western Blot for myogenin expression wereperformed. 2 )In animal study, animal model of alcoholism was made by alcoholic diet( 1 month ) then muscle ofposterior limb was harmed, the proliferation of satellite cells and formation of new muscle bundles were evaluated byhistological section. Results: The number of myotubes and expression of myogenin in experimental group were lessthan control group; but there was no significant difference on numbers of satellite cells and new muscle bundlesbetween two groups in animal model of alcoholism. Conclusion: Ethanol can inhibite differentiation of satelliteuscdecreasing of myogenin expressionI Key words muscle; satellite cells; ethanol; myogenin; differentiean中國煤化工CNMHG肌肉衛(wèi)星細(xì)胞位于骨骼肌纖維的肌細(xì)胞膜與基底膜之間史新的朊力,又被稱為肌肉干細(xì)胞收稿日期]2012-06-01修回日期]2012-06-15[基金項(xiàng)目]國家自然科學(xué)基金面上項(xiàng)目(81071480)作者簡方據(jù)1968-,男,江蘇南京人,主任醫(yī)師,醫(yī)學(xué)博土。Eml: anshen@ yahoo, cn沈干,等.乙醇對肌肉衛(wèi)星細(xì)胞 myogenin表達(dá)的影響675在肌肉形成以及肌肉損傷修復(fù)中扮演重要作用。慢性中貼壁培養(yǎng),每2日換液Ⅰ次,待細(xì)胞接近80%融合酒精中毒可引起肌肉萎縮,甚至引起酒精性心臟病。時進(jìn)行實(shí)驗(yàn)本硏究中作者對慢性酒精中毒過程中乙醇是否通過損1.2.3衛(wèi)星細(xì)胞向肌肉誘導(dǎo)分化將衛(wèi)星細(xì)胞培養(yǎng)害衛(wèi)星細(xì)胞而引起酒精性肌肉萎縮、降低衛(wèi)星細(xì)胞修液置換為誘導(dǎo)分化液(DMEM+2%馬血清),實(shí)驗(yàn)組分復(fù)損傷肌肉的能力作一探討?；囵B(yǎng)液中另加0.01%~1%的乙醇,分化培養(yǎng)1周。1資料和方法觀察兩組衛(wèi)星細(xì)胞形成肌管( myotube)的數(shù)量。由非本實(shí)驗(yàn)參與人員隨機(jī)選擇不同培養(yǎng)時間培養(yǎng)皿中至少1.1實(shí)驗(yàn)動物及分組3個光鏡下視野(×10),計(jì)算肌管的數(shù)量,并進(jìn)行統(tǒng)計(jì)日齡2~3 d sprag- Dowley(SD)大鼠及FVB成年學(xué)處理。小鼠來自查爾斯河實(shí)驗(yàn)室(美國,波土頓)。(1)體外1.2.4衛(wèi)星細(xì)胞形成肌管的檢測鑒定細(xì)胞及肌管實(shí)驗(yàn)分組:獲取大鼠肌肉衛(wèi)星細(xì)胞后實(shí)驗(yàn)分實(shí)驗(yàn)組以多聚甲醛在-20℃下固定1min,然后由PBS漂洗2乙醇作用組)及對照組(正常培養(yǎng)液對照組)。衛(wèi)星次;以10%馬血清封阻1h,然后加入一抗 myogenIn及細(xì)胞生長培養(yǎng)2d后更換為誘導(dǎo)分化液(DMEM+2%肌肉重鏈蛋白MHC(1:200小鼠單克隆抗體, Sigma公馬血清)誘導(dǎo)分化1周。實(shí)驗(yàn)組生長培養(yǎng)液及分化液司產(chǎn)品)作用半小時后,以PBS漂洗2次,加入二抗中加入1%的乙醇,共作用9d。對照組衛(wèi)星細(xì)胞培養(yǎng)(抗小鼠二抗連接免疫熒光Cy3, Sigma公司產(chǎn)品),作不加乙醇。(2)動物實(shí)驗(yàn)分組:FVB成年雄性小鼠,用半小時后,以PBS漂洗2次。細(xì)胞核以DAPI染色。體重20~30g,隨機(jī)分為兩組,每組10只。實(shí)驗(yàn)組給最后熒光顯微鏡觀察并拍照予乙醇飲食1個月;對照組給予正常飲食。然后檢測1.2.5 Western blot檢測乙醇對衛(wèi)星細(xì)胞 myogenIn表肌肉損傷后再生情況。達(dá)的影響于加入誘導(dǎo)液之前(0d)及誘導(dǎo)后的第1、1.2試劑及方法3、5、7d采取標(biāo)本,按 PIERCE公司Ec- Western Blot試1.2.1試劑胰蛋白酶( Sigma),Ⅱ型膠原酶(美國,劑盒操作說明書作 Western blot蛋白檢測。衛(wèi)星細(xì)胞wort- hington),F10培養(yǎng)液、胎牛血清(FBS)、DMEM及肌管裂解,蛋白電泳,轉(zhuǎn)膜,滴加 myogenin((1:200)培養(yǎng)液、馬血清均為 Gibco公司產(chǎn)品。ADS緩沖液由抗體,umi- Phos WB化學(xué)熒光素底物標(biāo)記,暗房X光Gorden huggins實(shí)驗(yàn)室提供。80μm尼龍網(wǎng)( Becton,片曝光。內(nèi)參照為 GAPDHDickinson and Company,美國),10m/2(C3H)細(xì)胞系由1.2.6乙醇對10Tl/2(C3H)細(xì)胞系的moD質(zhì)粒影Gorden huggins實(shí)驗(yàn)室提供,myo質(zhì)粒由 Gorden響小鼠胚胎間充質(zhì)干細(xì)胞10m2(C3H)細(xì)胞系轉(zhuǎn)Huggins實(shí)驗(yàn)室提供,鼠乙醇飼料( Dyets#710260,美染myoD質(zhì)粒。6孔板培養(yǎng)10T1/2細(xì)胞細(xì)胞數(shù)量為國),鼠對照正常飼料 Dyets,#710027,美國)。4×103,待細(xì)胞80%融合后,按照 Polyfectamine使用1.2.2肌肉衛(wèi)星細(xì)胞獲取將SD大鼠斷頸處死,以指南進(jìn)行操作,轉(zhuǎn)染myoD質(zhì)粒。培養(yǎng)2d后,更換為75%酒精消毒皮膚,取后腿肌肉,冷PBS沖洗數(shù)次,以含馬血清的分化培養(yǎng)液,作用2d后收取細(xì)胞, Western銳利眼科剪反復(fù)剪切肌肉組織,直至肌肉呈肉糜狀。Blωt檢測乙醇對1OT/2(C3H)細(xì)胞moD質(zhì)粒的表達(dá)加入終濃度為100U·m-Ⅱ型膠原酶及0.4mg·及 myogenIn表達(dá)的影響。ml-胰酶消化酶溶液,置入37℃水浴箱中,每15-1.2.7動物實(shí)驗(yàn)予FVB小鼠乙醇飲食1個月,對20min振蕩1次,消化30~45min,吸取消化液與細(xì)胞照組予正常飲食。然后制造鼠后肢肌肉損傷模型。具的混懸液,由80μm的尼龍濾網(wǎng)濾去肌腱等組織碎體操作如下:常規(guī)消毒,切開皮膚,分離后肢脛前肌,以片,對未消化的肌肉組織可加入0.05%胰酶,37℃水蚊式鉗橫行夾斷肌肉,同時將BrU藥片植入小鼠皮浴箱中再消化20min,不斷搖晃直至大部分肌肉組織下,在損傷后的第2天和第7天取肌肉標(biāo)本,固定后切被消化。同樣以80μm的尼龍濾網(wǎng)濾去未消化的組片,Br染色。光學(xué)顯微鏡下觀察肌束及BrU染色織碎片。陽性的衛(wèi)星細(xì)胞,計(jì)數(shù)并拍照。細(xì)胞混懸液離心5min(1200r·min-1),棄去上1.3統(tǒng)計(jì)學(xué)處理凊液,加入肌肉衛(wèi)星細(xì)胞培養(yǎng)液(F10培養(yǎng)液+20%實(shí)應(yīng)用spsS13.0統(tǒng)計(jì)軟件處理,實(shí)驗(yàn)組FBS),移入由Ⅳ型膠原預(yù)先處理的10cm的培養(yǎng)皿,與對中國煤化工細(xì)胞混懸液重新移入由Ⅳ型膠原預(yù)先處理的10cm的2結(jié)類 CNMHG"置入37℃5%CO2培養(yǎng)箱,靜置30~40min,然后吸取培養(yǎng)皿,再次進(jìn)行預(yù)貼壁,使混雜在衛(wèi)星細(xì)胞中的成纖2.1大鼠原代衛(wèi)星細(xì)胞培養(yǎng)維細(xì)胞盡量通過貼壁被祛除干凈。操作重復(fù)5~6次以倒置顯微鏡觀察可見,絕大多數(shù)先貼壁的細(xì)胞為得到純度幕棘肉衛(wèi)星細(xì)胞。放置在CO2培養(yǎng)箱成纖維細(xì)胞,細(xì)胞較大,而肌肉衛(wèi)星細(xì)胞體積較小,貼676東南大學(xué)學(xué)報(bào)(醫(yī)學(xué)版)2012年12月,31(6)壁速度較慢,仍處于懸浮狀態(tài)。經(jīng)反復(fù)預(yù)貼壁后,肌肉2.2肌肉衛(wèi)星細(xì)胞形成肌管的鑒定細(xì)胞消化懸浮液中成纖維細(xì)胞含量大大減少;靜置貼肌管免疫細(xì)胞化學(xué) myogenin及肌肉重鏈蛋白壁的為肌肉衛(wèi)星細(xì)胞,體積較成纖維細(xì)胞小,當(dāng)增殖速(MHC)反應(yīng)陽性,其中 myogenIn表達(dá)在細(xì)胞核,可見度快,培養(yǎng)密度較大時,容易發(fā)生細(xì)胞融合,形成肌管;多個細(xì)胞核排列在肌管中;而MHC表達(dá)在細(xì)胞漿中將培養(yǎng)液更換為含2%馬血清的分化培養(yǎng)液后肌管形(圖2)。成更充分(圖1)。2.3肌管計(jì)數(shù)、衛(wèi)星細(xì)胞數(shù)量和形態(tài)隨機(jī)計(jì)數(shù),可見實(shí)驗(yàn)組肌管數(shù)量(圖3)較對照組少,經(jīng)統(tǒng)計(jì)差異有統(tǒng)計(jì)學(xué)意義(P<0.05)。 myogenIn染色顯示衛(wèi)星細(xì)胞的數(shù)量和形態(tài)在誘導(dǎo)之前兩組無差異(圖4);在誘導(dǎo)分化第7天,兩組的肌管計(jì)數(shù)的差異有統(tǒng)計(jì)學(xué)意義(圖5)。2.4 Western Blot檢測乙醇對衛(wèi)星細(xì)胞 myogenIn表達(dá)的影響在細(xì)胞培養(yǎng)的不同時間點(diǎn)( Days Culture),經(jīng)乙醇作用衛(wèi)星細(xì)胞 myogenin表達(dá)逐漸減少(圖7)。雖然對照組 myogenIn表達(dá)也逐漸減少,但是實(shí)驗(yàn)組圖1肌肉衛(wèi)星細(xì)胞分化培養(yǎng)液誘導(dǎo)下形成的肌管Fig1 Myotubes formed by muscle satellite cells which induced myogenin表達(dá)下降更快,在誘導(dǎo)后第7天已幾乎檢測不by differential medium x 100Alexa 546DA鬥MereeMHCA.肌管內(nèi)多細(xì)胞核對 myogenin熒光反應(yīng)陽性;B.細(xì)胞核DAP染色;C.兩D.肌管MHC免疫熒光反應(yīng)陽性;E.DAP染色(下中);F.兩圖重疊TH中國煤化工CNMHG圖2肌管免疫細(xì)胞化學(xué)檢測A Nuclei of muscle cells are positive for myogenin; B. Staining of DAPl: C. merging of two picturesD Myotube is positive for MHC; E. Staining of DAPI( lower middle ) F. merging of two picturesFig 2 Immunocytochemistry for myotubes沈干,等.乙醇對肌肉衛(wèi)星細(xì)胞 myogenin表達(dá)的影響677一對照糾同樣以 Western blot檢測細(xì)胞myoD及 myogeNin表達(dá)實(shí)驗(yàn)糾(圖8),而mvoD表達(dá)實(shí)驗(yàn)組與對照組之間無顯著性差異,但實(shí)驗(yàn)組 myogeNin明顯表達(dá)下降。2.6肌肉損傷后新肌束形成及衛(wèi)星細(xì)胞的變化實(shí)驗(yàn)組在乙醇飲食1個月后,取小鼠后肢未損傷的肌肉切片,光鏡下觀察可見肌束邊衛(wèi)星細(xì)胞數(shù)量相對較對照組少(圖9),但差異無統(tǒng)計(jì)學(xué)意義(P>0.05)。小鼠肌肉損傷后2d,損傷部位切片可見肌束斷裂,衛(wèi)星細(xì)胞數(shù)量大量增加,新生衛(wèi)星細(xì)胞容易結(jié)合BrU,圖3肌管計(jì)數(shù)Br山U染色呈現(xiàn)陽性。但是對照組與實(shí)驗(yàn)組細(xì)胞數(shù)量Fig 3 Myotubes quantitation差異無統(tǒng)計(jì)學(xué)意義(P>0.05X圖10)2.510T1/2(C3H)轉(zhuǎn)染myoD質(zhì)粒后myoD及在后肢損傷后7d,對照組與實(shí)驗(yàn)組損傷部位的肌肉切片,可見許多新形成的肌束,肌束內(nèi)成串的細(xì)胞myogenIn表達(dá)細(xì)胞系10T2C3H)轉(zhuǎn)染myoD質(zhì)粒后,誘導(dǎo)2d核,與體外衛(wèi)星細(xì)胞融合的狀態(tài)相同(圖11)。圖4誘導(dǎo)分化前對照組(A)與實(shí)驗(yàn)組(B)細(xì)胞 myogenin熒光染色Fig 4 Before differentiation, myogenin staining for cells in control group( A )and exporimental group( B中國煤化工圖5誘導(dǎo)分化1周對照組(A)與實(shí)驗(yàn)組( B )myogenin熒光染色YHCNMHGFig 5 Myogenin staining for myotube after cultured in differentiation medium for 1 week, control group( A ) and experimentalgroup678東南大學(xué)學(xué)報(bào)(醫(yī)學(xué)版)2012年12月,31(6)圖6誘導(dǎo)分化1周對照組(A)與實(shí)驗(yàn)組(BMHC熒光染色Fig 6 MHC staining for myotube of control group( a )and experimental group( B )after cultured in differentiation medium for 1b a b a b laelMyogenGAPDH圖7 Western blot顯示不同時間點(diǎn)(0~7d)實(shí)驗(yàn)組(a)與對照組( b)myogenin表達(dá)的變化, GAPDH為內(nèi)參照Fig 7 Western Blot showed expression of myogenin in experimental group( a and control group( b )in different timeints( 1- 7 daysGAPDH圖8細(xì)胞系10T2C3H)轉(zhuǎn)染myoD后,誘導(dǎo)2d后, Western中國煤化工傷的肌肉切片,肌束邊可Blot顯示實(shí)驗(yàn)組a)及對照組 b)myoD及 myogenin表達(dá)Fig8 MyoD was transfected into 10T1/2( C3H ) then culturedCNMHInJuried muscle in control group( A )andrentiation medium Western Blot showed myoD andexperimental group( B ) satellite cells were beside of myotubesmyogenin expression in ethanol group a )and control group bHE×100沈干,等.乙醇對肌肉衛(wèi)星細(xì)胞 myogenin表達(dá)的影響679線圖10對照組(A)與實(shí)驗(yàn)組(B)損傷第2天的肌肉切片HE×100Fig 10 Section of injuried muscle( Day 2)in control group( A )and experimental group B ) the broken myotubes and increasednumber of satellite cells were shown HE x 100圖11對照組(A)與實(shí)驗(yàn)組(B損傷第7天的肌肉切片HE×200Fig 11 Section of injuried muscle( Day 7)in control group( A) and experimental group B ), many myotubes were newly formedand many neuclei were arranged in chain HE X 2003討論肌肉衛(wèi)星細(xì)胞只存在于骨骼肌內(nèi),心肌和平滑肌內(nèi)沒有衛(wèi)星細(xì)胞。隨著年齡的增加,衛(wèi)星細(xì)胞的密度長期酒精攝入是骨骼肌和心肌萎縮的主要原因。逐漸減少,到一定程度后,終身維持。在出生后一段時乙醇或其代謝產(chǎn)物對細(xì)胞均有毒性作用:乙醇脫氫酶間內(nèi),衛(wèi)星細(xì)胞核的比例明顯下降,這主要是因?yàn)殡S著可使乳酸產(chǎn)生,改變細(xì)胞的代謝;乙醇可以使線粒體膨衛(wèi)星細(xì)胞的融合,而肌細(xì)胞核總數(shù)增加23。所以我大,脆性增加,引起線粒體的損害。本實(shí)驗(yàn)?zāi)康氖翘剿鱾儾捎贸錾?~3d的大鼠,其肌肉組織中富含肌肉衛(wèi)乙醇是否可通過損害肌肉衛(wèi)星細(xì)胞而引起肌肉的萎縮星細(xì)胞。我們也曾以單肌纖維貼壁方法4培養(yǎng)出衛(wèi)和損傷,或者影響肌肉損傷的修復(fù)。星細(xì)胞(結(jié)果未顯示),但由于技術(shù)要求較高,衛(wèi)星細(xì)肌肉衛(wèi)星細(xì)胞又被稱為肌肉干細(xì)胞,位于骨骼肌胞的產(chǎn)量有限。所以在需要大量衛(wèi)星細(xì)胞的實(shí)驗(yàn)中我纖維的肌細(xì)胞膜與基底膜之間,其核較大、胞漿少,細(xì)們采用幼鼠肉糜消化法。胞器數(shù)量少。在損傷等情況下,衛(wèi)星細(xì)胞被激活,分裂肌源性決定因子(MDF)在肌細(xì)胞分化中均發(fā)揮關(guān)增殖,參與肌肉的修復(fù)。鍵作用,這些因子包括M5、M6、 myogenIn和myoD肌肉衛(wèi)星細(xì)胞具有自我更新的能力,以維持其數(shù)而 myogenIn是由Myf5和myoD激活的。在分子水平量的大體穩(wěn)定。肌肉衛(wèi)星細(xì)胞的自我更新通過兩種機(jī)靜止期細(xì)陷油謝的牛征是M5和myoD的制:一是由非對稱性分裂所造成的,大部分的子細(xì)胞定迅速型于肌源性分化而另一部分的子細(xì)胞重新成為衛(wèi)星M5中國煤化工早表達(dá)上調(diào),隨后出現(xiàn)CNMHGgenin及MHC是肌肉細(xì)胞閣;二是肌肉衛(wèi)星細(xì)胞進(jìn)行對稱性分裂,其中一個分化晚期表達(dá)的蛋白56。剛從肌纖維分離出的衛(wèi)星細(xì)激活衛(wèi)星細(xì)胞可以退出細(xì)胞循環(huán),重新進(jìn)人靜止?fàn)顟B(tài),胞并不表達(dá)肌源性標(biāo)志物My5和myoD,提示靜止期衛(wèi)并可以被再次激活進(jìn)人細(xì)胞循環(huán),以進(jìn)行衛(wèi)星細(xì)胞的星細(xì)胞更具原始性。所以我們主要以 myogenin為肌更新11肉標(biāo)志,檢測衛(wèi)星細(xì)胞及肌管的肌肉特性680東南大學(xué)學(xué)報(bào)(醫(yī)學(xué)版)2012年12月,31(6)本研究發(fā)現(xiàn),乙醇對衛(wèi)星細(xì)胞 myogenin表達(dá)有顯著影響,而對myoD影響不顯著。說明其作用的位點(diǎn)[參考文獻(xiàn)]在肌肉發(fā)育的較遲階段,使衛(wèi)星細(xì)胞形成肌管數(shù)量較1] CONBOY IM, RANDOT A, The regulation of Notch signaling對照組少。在 Western blot檢測中兩組 myogenIn表達(dá)controls satellite cell activation and cell fate determination in也有差異。我們以myo質(zhì)粒轉(zhuǎn)染細(xì)胞系10T1/2postnatal myogenesis J ]. Dev Cell, 2002, 3( 3):397-409(C3H)細(xì)胞,誘導(dǎo)細(xì)胞向肌細(xì)胞分化,并同樣以乙醇2 GIBSON M C, SCHULTZ E. The distribution of satellite cells處理,也發(fā)現(xiàn)乙醇在作用2d后使 myogenin表達(dá)下降and their relationship to specific fiber types in soleus and ex-tensor digitorum longus muscles J]. Anat Rec, 1982, 202(3)而對myoD表達(dá)影響不大。329-337乙醇會對身體帶來多方面的影響,甚至可以影[3] IBSON M C, SCHULTZ E. Age-related differences inabsolut響人間充質(zhì)干細(xì)胞的分化9。本實(shí)驗(yàn)也證實(shí)了乙醇lumbers of skeletal muscle satellite cells[ J] Muscle Nerv對肌肉干細(xì)胞的負(fù)面影響。為進(jìn)一步驗(yàn)證乙醇對在體1983,6(8):574580肌肉的影響,我們給予實(shí)驗(yàn)動物乙醇飲食,并且在乙醇4] ROSENBLATT DJ, LUNT A, PARRY D J,etal. Culturing of飲食后1個月后制造肌肉損傷模型。而且處于增殖和satellite cells from living single muscle fibers explants[ J ].In分裂的細(xì)胞對BrdU染色顯示陽性,肌肉的衛(wèi)星細(xì)胞Vitro Cell Dev Biol Anim. 1995.31( 10): 773-779在肌肉損傷的情況下能夠被激活。在肌肉損傷后2[5 COOPER R N, TAJBAKHAH S, MOULY V, et al. In vivo satel通過組織切片,我們發(fā)現(xiàn)衛(wèi)星細(xì)胞數(shù)量大大增加;在損lite cell activation via Myf5 and MyoD in regeneration mouseskeletal muscle JI J Cell Sci, 1999, 112( Pt 17): 2895-2901傷后7d,這些衛(wèi)星細(xì)胞融合形成新的肌束,肌束內(nèi)細(xì)[61wEQ, PATERSON B M. Regulation of MyoD function in the胞核排列成串,與體外衛(wèi)星細(xì)胞融合的表現(xiàn)一樣。但dividing myoblast J ]. FEBS Lett, 2001, 490(3): 171-178可能由于動物乙醇飲食的時間較短,沒有完全體現(xiàn)乙[7] YABLONKA RZ, RUDNICKIM A, RIVERA A J,etal.The醇的慢性中毒損傷,所以實(shí)驗(yàn)組與對照組在衛(wèi)星細(xì)胞transition from proliferation to differentiation is delayed in sat-的數(shù)量以及形成的新肌束數(shù)量上差異無統(tǒng)計(jì)學(xué)意義ellite cells from mice lacking MyoD[ J ]. Dev Biol, 1999, 210我們也對損傷肌肉進(jìn)行了PCR等方法檢測損傷肌肉(2):440-45myogenin表達(dá)(結(jié)果未顯示),亦未得到兩組有顯著差81HPA.HPD, ATALA A, et al. Ethanol alters the osteo-異的結(jié)果。在將來進(jìn)一步的實(shí)驗(yàn)中,我們將延長乙醇飲食的時間,真正模擬慢性乙醇中毒的情形。Alcohol Clin Exp Res, 2010, 34(10): 1714-1722本實(shí)驗(yàn)首次通過對衛(wèi)星細(xì)胞的培養(yǎng)來驗(yàn)證乙醇礦9JG0NGZ, WEZEMAN F H. Inhibitory effect of alcohol on osneogenic differentiation in human bor其向肌肉分化的影響,結(jié)果顯示乙醇可使衛(wèi)星細(xì)胞chymal stem cells[ J ] Alcohol Clin Exp Res, 2004, 28(3myogenIn表達(dá)下降,從而抑制其肌性分化。468-479《現(xiàn)代醫(yī)學(xué)》簡介《現(xiàn)代醫(yī)學(xué)》是由教育部主管、東南大學(xué)主辦的國家級核心期刊,1964年創(chuàng)刊,現(xiàn)已出版發(fā)行40卷。2013年由雙月刊改為月刊。《現(xiàn)代醫(yī)學(xué)》為中國科技論文統(tǒng)計(jì)源期刊,即中國科技核心期刊,多年來,《現(xiàn)代醫(yī)學(xué)》一直被《中國核心期刊遴選)數(shù)據(jù)庫》、《中國學(xué)術(shù)期刊綜合評價數(shù)據(jù)庫》、《中國科學(xué)引文數(shù)據(jù)庫》、《中國期刊網(wǎng)》、《中國學(xué)術(shù)期刊光盤版)》、《中文科技期刊數(shù)據(jù)庫》、《資源系統(tǒng)》、《天元數(shù)據(jù)網(wǎng)》《龍?jiān)雌诳W(wǎng)》、《中國教育閱讀網(wǎng)》全文收錄,同時被《中國醫(yī)學(xué)文摘》《中國藥學(xué)文摘》各分冊收錄。從2011年3月開始和中國學(xué)術(shù)期刊(光盤版)電子雜志社正式簽署了獨(dú)家數(shù)字出版合作協(xié)議。《現(xiàn)代醫(yī)學(xué)》向國內(nèi)外公開發(fā)行,國際標(biāo)準(zhǔn)刊號:ISSN1671-7562、國內(nèi)統(tǒng)一刊號:CN32-1659/R。其以中高級衛(wèi)生工作者為主要讀者對象,主要報(bào)道醫(yī)療衛(wèi)生工作者在臨床醫(yī)學(xué)影像醫(yī)學(xué)等方面的先進(jìn)經(jīng)驗(yàn)、科研成果以及與實(shí)踐密切結(jié)合的基礎(chǔ)醫(yī)學(xué)硏究。設(shè)有專家論壇Fu中國煤化工論綜述個案報(bào)道、誤診誤治、中醫(yī)中藥、護(hù)理等欄目CNMHG自2009年起現(xiàn)代醫(yī)學(xué)啟用網(wǎng)上投稿系統(tǒng)(網(wǎng)址hp://w.xdyx.org.cn),作者可登陸后注冊投稿,并跟蹤稿件狀態(tài)。通信地址:江蘇省南京市丁家橋87號;郵編:210009;電話:025-83272479,83272481,83272483。E-m亙方數(shù)提@pub.seu,cd,cm