中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2009年
12期
921-924
,共4页
季菊玲%张锦生%王雪晴%季煜华
季菊玲%張錦生%王雪晴%季煜華
계국령%장금생%왕설청%계욱화
肝星状细胞%蛋白质组%串联质谱分析%二维纳升高效液相
肝星狀細胞%蛋白質組%串聯質譜分析%二維納升高效液相
간성상세포%단백질조%천련질보분석%이유납승고효액상
Hepatic stellatecell%Proteome%Tandem mass spectrometry%Two dimensional nano liquid chromatography
目的 用在线二维纳升高效液相结合串联质谱(2D nanoLC-MS/MS)技术分析体外培养大鼠原代肝星状细胞(HSC)的蛋白质表达谱. 方法 分离培养大鼠原代HSC,取体外培养第10天自动活化的大鼠HSC总蛋白质,通过2D nanoLC-MS/MS对酶解所获肽段进行分离和鉴定,并对鉴定的蛋白质进行细胞定位和功能分类. 结果从50μg HSC总蛋白质中鉴定出1014种蛋白质,相对分子质量范围为7832～588 364;主要分布于细胞核、细胞骨架、线粒体、内质网,分别占25%,17%、13%、13%.蛋白质功能主要与核酸代谢,细胞器组装、信号传导、能量代谢等生理进程有关,分别占17%、14%,10%、9%;其中细胞骨架蛋白α-平滑肌肌动蛋白,波形蛋白和结蛋白是活化大鼠HSC特异性表达的蛋白质.结论 本研究获得了目前较为全面的活化大鼠原代HSC的蛋白质表达谱,为深入研究HSC的活化机制奠定了基础.
目的 用在線二維納升高效液相結閤串聯質譜(2D nanoLC-MS/MS)技術分析體外培養大鼠原代肝星狀細胞(HSC)的蛋白質錶達譜. 方法 分離培養大鼠原代HSC,取體外培養第10天自動活化的大鼠HSC總蛋白質,通過2D nanoLC-MS/MS對酶解所穫肽段進行分離和鑒定,併對鑒定的蛋白質進行細胞定位和功能分類. 結果從50μg HSC總蛋白質中鑒定齣1014種蛋白質,相對分子質量範圍為7832～588 364;主要分佈于細胞覈、細胞骨架、線粒體、內質網,分彆佔25%,17%、13%、13%.蛋白質功能主要與覈痠代謝,細胞器組裝、信號傳導、能量代謝等生理進程有關,分彆佔17%、14%,10%、9%;其中細胞骨架蛋白α-平滑肌肌動蛋白,波形蛋白和結蛋白是活化大鼠HSC特異性錶達的蛋白質.結論 本研究穫得瞭目前較為全麵的活化大鼠原代HSC的蛋白質錶達譜,為深入研究HSC的活化機製奠定瞭基礎.
목적 용재선이유납승고효액상결합천련질보(2D nanoLC-MS/MS)기술분석체외배양대서원대간성상세포(HSC)적단백질표체보. 방법 분리배양대서원대HSC,취체외배양제10천자동활화적대서HSC총단백질,통과2D nanoLC-MS/MS대매해소획태단진행분리화감정,병대감정적단백질진행세포정위화공능분류. 결과종50μg HSC총단백질중감정출1014충단백질,상대분자질량범위위7832～588 364;주요분포우세포핵、세포골가、선립체、내질망,분별점25%,17%、13%、13%.단백질공능주요여핵산대사,세포기조장、신호전도、능량대사등생리진정유관,분별점17%、14%,10%、9%;기중세포골가단백α-평활기기동단백,파형단백화결단백시활화대서HSC특이성표체적단백질.결론 본연구획득료목전교위전면적활화대서원대HSC적단백질표체보,위심입연구HSC적활화궤제전정료기출.
Objective To profile the protein expression in activated rat hepatic stenate cells(HSCs).Methods Primary rat HSCs were isolated and cultured in vitro.After 10 days in vitro culture,the HSCs were activated.Total protein exffacted from these activated HSCs were digested.and the obtained peptides were analyzed by using online 2D nanoLC-MS/MS.The identified proteins were classified according to their distrbutions and functions.Results 1014 proteins were identified from 50 μg HSCs protein extract,the molecular weights of these proteins ranged from 7832 Da to 588 364 Da.Most of these proteins resided in nucleus,cytoskeleton,mitochondrion and endoplasmic reticulum.And these proteins were mainly involved in nucleic acid metobolism,organelle organization,signal transduction and energy generation.Among these proteins,alpha-smooth muscle actin,vimentin and desmin were specifically expressed in activated HSCs.Conclusion To the best of our knowledge,this is the most comprehensive protein expression profile of activated rat HSCs.
