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[1]史曉林,楊依然,劉鐘,等.基于蛋白質(zhì)組學(xué)方法分析絕經(jīng)后骨質(zhì)疏松癥全血差異蛋白[J].中醫(yī)正骨,2019,31(06):7-11.
　SHI Xiaolin,YANG Yiran,LIU Zhong,et al.Analysis of proteins which are differentially expressed in whole blood of patients with postmenopausal osteoporosis using proteomics approach[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2019,31(06):7-11.
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基于蛋白質(zhì)組學(xué)方法分析絕經(jīng)后骨質(zhì)疏松癥全血差異蛋白()
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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第31卷
期數(shù):
2019年06期
頁(yè)碼:
7-11
欄目:
臨床研究
出版日期:
2019-06-20

文章信息/Info

Title:
Analysis of proteins which are differentially expressed in whole blood of patients with postmenopausal osteoporosis using proteomics approach
作者:
史曉林1; 楊依然2; 劉鐘2; 李春雯3; 梁博程1; 劉康1; 何偉濤4; 胡炯1
(1.浙江中醫(yī)藥大學(xué)附屬第二醫(yī)院,浙江 杭州 310005; 2.浙江中醫(yī)藥大學(xué)第二臨床醫(yī)學(xué)院,浙江 杭州 310053; 3.浙江中醫(yī)藥大學(xué)基礎(chǔ)醫(yī)學(xué)院,浙江 杭州 310053; 4.海鹽縣中醫(yī)院,浙江 海鹽 314300)
Author(s):
SHI Xiaolin1; YANG Yiran2; LIU Zhong2; LI Chunwen3; LIANG Bocheng1; LIU Kang1; HE Weitao4; HU Jiong1
1.The Second Affiliated Hospital of Zhejiang Chinese Medical University,Hangzhou 310005,Zhejiang,China; 2.The Second Clinical Medical College of Zhejiang Chinese Medical University,Hangzhou 310053,Zhejiang,China; 3.The basic Medical College of Zhejiang Chinese Medical University,Hangzhou 310053,Zhejiang,China; 4.Traditional Chinese Medical Hospital of Haiyan county,Haiyan 314300,Zhejiang,China
關(guān)鍵詞:
骨質(zhì)疏松; 絕經(jīng)后;  蛋白質(zhì)組學(xué);  色譜法;  高壓液相;  串聯(lián)質(zhì)譜法;  差異蛋白
Keywords:
osteoporosis; postmenopausal;  proteomics;  chromatography; high pressure liquid;  tandem mass spectrometry;  differentially expressed protein
摘要:
目的:基于蛋白質(zhì)組學(xué)方法篩選絕經(jīng)后骨質(zhì)疏松癥(postmenopausal osteoporosis,PMOP)全血差異蛋白。方法:選擇7例PMOP患者(骨質(zhì)疏松組)和7例絕經(jīng)后骨量正常女性(骨量正常組)作為研究對(duì)象。空腹抽取靜脈血,采用高效液相色譜-串聯(lián)質(zhì)譜技術(shù)篩選PMOP全血差異蛋白,選取其中發(fā)生顯著變化的差異蛋白進(jìn)行生物信息學(xué)分析。結(jié)果:①差異蛋白鑒定及篩選結(jié)果。應(yīng)用高效液相色譜-串聯(lián)質(zhì)譜技術(shù)共定量檢測(cè)到102個(gè)PMOP差異蛋白質(zhì),其中25個(gè)蛋白質(zhì)位點(diǎn)表達(dá)上調(diào)、77個(gè)蛋白質(zhì)位點(diǎn)表達(dá)下調(diào)。68個(gè)蛋白質(zhì)兩組差異倍數(shù)超過2.0,其中39個(gè)蛋白質(zhì)兩組差異倍數(shù)有統(tǒng)計(jì)學(xué)意義(P<0.01)。②生物信息學(xué)分析結(jié)果。基因功能注釋發(fā)現(xiàn),差異蛋白主要參與的細(xì)胞活動(dòng)為單一生物過程、細(xì)胞代謝過程、細(xì)胞生物調(diào)節(jié)、對(duì)刺激和代謝過程的反應(yīng),主要的分子功能為催化活性和結(jié)構(gòu)分子活動(dòng)。亞細(xì)胞定位分析發(fā)現(xiàn),102個(gè)差異蛋白質(zhì)中,位點(diǎn)表達(dá)上調(diào)的25個(gè)差異蛋白全部位于細(xì)胞內(nèi),位點(diǎn)表達(dá)下調(diào)的77個(gè)差異蛋白中,19個(gè)在細(xì)胞外,其他均在細(xì)胞內(nèi)。39個(gè)目標(biāo)差異蛋白相互作用網(wǎng)絡(luò)圖顯示,蛋白酶體亞基α型-7(proteasome subunit alpha type-7,PSMA7)、蛋白酶體亞基β型-7(proteasome subunit beta type-7,PSMB7)、蛋白酶體亞基β型-3(proteasome subunit beta type-3,PSMB3)、蛋白酶體亞基α型-1(proteasome subunit alpha type-1,PSMA1)、神經(jīng)前體細(xì)胞表達(dá)發(fā)育下調(diào)因子8(neural precursor cell-expressed developmentally downregulated 8,NEDD8)、DNA損傷結(jié)合蛋白1(DNA damage-binding protein 1,DDB1)、RAD23A、UBA52等蛋白位于相互作用網(wǎng)絡(luò)節(jié)點(diǎn)的中心。結(jié)論:以全血作為樣本采用蛋白質(zhì)組學(xué)方法可篩選出數(shù)量豐富的PMOP差異蛋白,其中PSMA7、PSMB7、PSMB3、PSMA1、NEDD8、DDB1、RAD23A、UBA52可能是PMOP的潛在有效血液標(biāo)志物。
Abstract:
Objective:To select the proteins which are differentially expressed in whole blood of patients with postmenopausal osteoporosis(PMOP)using proteomics approach.Methods:Seven females with PMOP(osteoporosis group)and seven postmenopausal females with normal bone mass(normal bone mass group)were selected out as the subjects.The fasting blood were fetched out from vein.The proteins which were differentially expressed in whole blood of patients with PMOP were selected out by using high pressure liquid chromatography-tandem mass spectrometry(HPLC-MS/MS),and those proteins which were obviously differentially expressed in whole blood were picked out for further bioinformatic analysis.Results:A total of 102 differentially expressed proteins were found out by HPLC-MS/MS,in which the expressions of 25 protein loci were up-regulated and the expressions of 77 protein loci were down-regulated.The fold change of 68 proteins exceeded 2.0,while there was statistical significance in fold change of 39 out of 68 proteins(P<0.01).The results of gene annotation demonstrated that the differentially expressed proteins were mainly involved in 4 kinds of cell activities,including single biological process,cell metabolism process,cell biological regulation and response to stimulation and metabolism process,and their main molecular functions included catalytic activities and structural molecular activities.The results of subcellular positioning analysis demonstrated that 25 differentially expressed proteins with up-regulated loci expressions were inside the cells,and out of 77 differentially expressed proteins with down-regulated loci expressions 19 were outside the cells and the others were inside the cells.The network diagram of interaction between 39 differentially expressed proteins showed that proteasome subunit alpha type-7(PSMA7),proteasome subunit beta type-7(PSMB7),proteasome subunit beta type-3(PSMB3),proteasome subunit alpha type-1(PSMA1),neural precursor cell-expressed developmentally downregulated 8(NEDD8),DNA damage-binding protein 1(DDB1),RAD23A and UBA52 were located at the center of the interaction network nodes.Conclusion:Abundant differentially expressed proteins of PMOP in whole blood can be identified using proteomics approach,and PSMA7,PSMB7,PSMB3,PSMA1,NEDD8,DDB1,RAD23A and UBA52 may be the potential effective blood markers of PMOP.

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備注/Memo

備注/Memo:
基金項(xiàng)目:國(guó)家自然科學(xué)基金項(xiàng)目(81573754; 81873129); “十三五”浙江省中醫(yī)藥(中西醫(yī)結(jié)合)重點(diǎn)學(xué)科建設(shè)計(jì)劃項(xiàng)目(2017-XK-A16) 通訊作者:史曉林 E-mail:[email protected](收稿日期:2019-04-06 本文編輯:李曉樂)
更新日期/Last Update: 1900-01-01