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《阻斷tgf-βtgf-βri通路對(duì)atdc5細(xì)胞增殖,分化及細(xì)胞外基質(zhì)的影響》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、阻斷TGF-β/TGF-βRI通路對(duì)ATDC5細(xì)胞增殖、分化及細(xì)胞外基質(zhì)的影響王權(quán)，戚華兵，王曉鳳，朱瑩，陳林（400042重慶，第三軍醫(yī)大學(xué)大坪醫(yī)院野戰(zhàn)外科研究所，創(chuàng)傷實(shí)驗(yàn)室，骨代謝與修復(fù)中心，創(chuàng)傷、燒傷與復(fù)合傷國(guó)家重點(diǎn)實(shí)驗(yàn)室）[摘要]目的研究TGF-βRI阻斷劑SB-505124對(duì)軟骨細(xì)胞增殖、分化及細(xì)胞外基質(zhì)的影響與機(jī)制。方法利用Westernblot檢測(cè)不同濃度的SB-505124處理后，前軟骨細(xì)胞系A(chǔ)TDC5中p-Smad2/3的變化。MTT法檢測(cè)SB-505124處理對(duì)ATDC5細(xì)胞生長(zhǎng)、增殖的影響及時(shí)間、濃度依賴(lài)性效應(yīng)。Western
2、blot檢測(cè)c-Myc蛋白水平的變化。Real-timePCR檢測(cè)SB-505124處理后軟骨分化、細(xì)胞外基質(zhì)合成及降解相關(guān)分子CollagenⅡ、Aggrecan及Adamts5表達(dá)與變化，并利用阿爾新藍(lán)染色法檢測(cè)SB-505124處理對(duì)軟骨細(xì)胞系蛋白聚糖合成的影響。結(jié)果Westernblot檢測(cè)示，SB-505124處理ATDC5細(xì)胞明顯抑制TGF-β1激活的p-Smad2/3，并呈濃度依賴(lài)性；MTT檢測(cè)示，SB-505124濃度和時(shí)間依賴(lài)性的抑制ATDC5細(xì)胞增殖；Real-timePCR檢測(cè)示，SB-505124處理明顯抑制ATDC5細(xì)胞中
3、CollagenⅡ、Aggrecan的表達(dá)，同時(shí)上調(diào)了Adamts5的表達(dá)；阿爾新藍(lán)染色則提示SB-505124處理明顯抑制ATDC5細(xì)胞中蛋白聚糖的合成。結(jié)論利用SB-505124阻斷內(nèi)源性TGF-β信號(hào)，可抑制軟骨細(xì)胞增殖、分化及細(xì)胞外基質(zhì)的合成，同時(shí)促進(jìn)細(xì)胞外基質(zhì)的降解。[關(guān)鍵詞]TGF-βRI；SB-505124；ATDC5；增殖；細(xì)胞外基質(zhì)[中圖法分類(lèi)號(hào)][文獻(xiàn)標(biāo)志碼]A[基金項(xiàng)目]國(guó)家自然科學(xué)基金重點(diǎn)項(xiàng)目（81030036）[通信作者]陳林，E-mail：linchen70@163.comEffectofinhibitingTGF-β/
4、TGF-βRIsignalingonproliferation,differentiationandextracellularmatrixofATDC5cellWangQuan,QiHuabing,WangXiaofeng,ZhuYing,ChenLin（StateKeyLaboratoryofTrauma,BurnsandCombinedInjury,CenterofBoneMetabolismandRepair,InstituteofSurgeryReserch,DapingHospital,ThirdMilitrayMedicalUniver
5、sity,Chongqing,400042,China）[Abstract]ObjectiveToinvestigatetheeffectofTGF-βRIinhibitorSB-505124onproliferation,differentiationandextracellularmatrixofchondrocytes.MethodsTheexpressionofp-Smad2/3inATDC5cellswhichweretreatedwithdifferentconcentrationofSB-505124wasanalyzedbywest
6、ernblot.TheeffectofSB-505124onproliferationofATDC5anditsrelationshipwithconcentrationwasdetectedbyMTTassay.TheeffectofSB-505124ontheexpressionofc-Mycproteinwasanalyzedbywesternblot.TheeffectsofSB-505124onthemarksofdifferentiationandextracellularmatrix,includingCollagenⅡ,Aggrec
7、anandAdamts5,wereanalysedbyReal-timePCR.TheeffectofSB-505124onexpressionofsulfatedproteoglycanswasmeasuredbyAlcianbluestaining.ResultsWesternBlotshowedthattreatmentwithSB-505124causedadeclineofphosphorylatedSmad2/3(p-Smad2/3),includingcarboxylterminiandlinkerregionstimulatedby
8、TGF-β1inATDC5cells.MTTassaydemonstratedthatSB-505124significa
