中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2014年
42期
3304-3309
,共6页
苏瑾文%刘艳华%杨秉芬%程小星
囌瑾文%劉豔華%楊秉芬%程小星
소근문%류염화%양병분%정소성
结核,肺%基因表达谱%信号通路%聚合酶链反应
結覈,肺%基因錶達譜%信號通路%聚閤酶鏈反應
결핵,폐%기인표체보%신호통로%취합매련반응
Tuberculosis,pulmonary%Gene expression profiling%Pathway%Polymerase chain reaction
目的 分析和验证重症继发性肺结核患者基因表达谱.方法 收集解放军三○九医院结核病研究所2012年5月至2013年10月住院患者中符合入选标准的继发性肺结核患者103例,根据病情严重程度分为重症组(57例)和轻症组(46例),并以同期进行健康体检且年龄相匹配的45名健康人为对照组.选取重症组4例、轻症组3例和对照组5名用Affymetrix基因表达谱芯片进行全基因组检测和分析,对重症组比对轻症组筛选的差异基因做聚类分析和生物信息学分析;对以上各组剩余的53例、43例和40名用实时荧光定量聚合酶链反应(RT-PCR)验证芯片结果,其中验证代表基因原癌基因(JUN)表达水平是重症组、轻症组各20例,对照组8名;用方差分析和非参数检验统计方法判断组间的统计学差异.结果 芯片结果显示重症组比对轻症组差异基因为406条,下调基因264条,上调基因142条,差异基因以表达下调为主;聚类分析显示同组样本表达谱有相似性,提示芯片试验有一定重复性,结果可靠;基因本体论注释显示差异基因主要参与了生物学过程,包括免疫反应、信号转导、DNA依赖的转录调节、炎症反应、抗原处理与提呈、趋化作用等;信号通路分析显示差异基因参与了22条炎症和免疫反应相关通路,主要有B细胞受体信号通路、抗原处理与提呈、Toll样受体信号通路、丝裂原活化蛋白激酶通路、转化生长因子-β通路等;RT-PCR验证JUN吸光度值在组间比较的统计学结果为重症组比轻症组P ＜0.001,重症组表达下调,与表达谱芯片结果相符.结论 重症继发性肺结核与轻症继发性肺结核患者相比对有大量差异表达基因,其肺结构严重损害与基因水平差异有关.
目的 分析和驗證重癥繼髮性肺結覈患者基因錶達譜.方法 收集解放軍三○九醫院結覈病研究所2012年5月至2013年10月住院患者中符閤入選標準的繼髮性肺結覈患者103例,根據病情嚴重程度分為重癥組(57例)和輕癥組(46例),併以同期進行健康體檢且年齡相匹配的45名健康人為對照組.選取重癥組4例、輕癥組3例和對照組5名用Affymetrix基因錶達譜芯片進行全基因組檢測和分析,對重癥組比對輕癥組篩選的差異基因做聚類分析和生物信息學分析;對以上各組剩餘的53例、43例和40名用實時熒光定量聚閤酶鏈反應(RT-PCR)驗證芯片結果,其中驗證代錶基因原癌基因(JUN)錶達水平是重癥組、輕癥組各20例,對照組8名;用方差分析和非參數檢驗統計方法判斷組間的統計學差異.結果 芯片結果顯示重癥組比對輕癥組差異基因為406條,下調基因264條,上調基因142條,差異基因以錶達下調為主;聚類分析顯示同組樣本錶達譜有相似性,提示芯片試驗有一定重複性,結果可靠;基因本體論註釋顯示差異基因主要參與瞭生物學過程,包括免疫反應、信號轉導、DNA依賴的轉錄調節、炎癥反應、抗原處理與提呈、趨化作用等;信號通路分析顯示差異基因參與瞭22條炎癥和免疫反應相關通路,主要有B細胞受體信號通路、抗原處理與提呈、Toll樣受體信號通路、絲裂原活化蛋白激酶通路、轉化生長因子-β通路等;RT-PCR驗證JUN吸光度值在組間比較的統計學結果為重癥組比輕癥組P ＜0.001,重癥組錶達下調,與錶達譜芯片結果相符.結論 重癥繼髮性肺結覈與輕癥繼髮性肺結覈患者相比對有大量差異錶達基因,其肺結構嚴重損害與基因水平差異有關.
목적 분석화험증중증계발성폐결핵환자기인표체보.방법 수집해방군삼○구의원결핵병연구소2012년5월지2013년10월주원환자중부합입선표준적계발성폐결핵환자103례,근거병정엄중정도분위중증조(57례)화경증조(46례),병이동기진행건강체검차년령상필배적45명건강인위대조조.선취중증조4례、경증조3례화대조조5명용Affymetrix기인표체보심편진행전기인조검측화분석,대중증조비대경증조사선적차이기인주취류분석화생물신식학분석;대이상각조잉여적53례、43례화40명용실시형광정량취합매련반응(RT-PCR)험증심편결과,기중험증대표기인원암기인(JUN)표체수평시중증조、경증조각20례,대조조8명;용방차분석화비삼수검험통계방법판단조간적통계학차이.결과 심편결과현시중증조비대경증조차이기인위406조,하조기인264조,상조기인142조,차이기인이표체하조위주;취류분석현시동조양본표체보유상사성,제시심편시험유일정중복성,결과가고;기인본체론주석현시차이기인주요삼여료생물학과정,포괄면역반응、신호전도、DNA의뢰적전록조절、염증반응、항원처리여제정、추화작용등;신호통로분석현시차이기인삼여료22조염증화면역반응상관통로,주요유B세포수체신호통로、항원처리여제정、Toll양수체신호통로、사렬원활화단백격매통로、전화생장인자-β통로등;RT-PCR험증JUN흡광도치재조간비교적통계학결과위중증조비경증조P ＜0.001,중증조표체하조,여표체보심편결과상부.결론 중증계발성폐결핵여경증계발성폐결핵환자상비대유대량차이표체기인,기폐결구엄중손해여기인수평차이유관.
Objective To explore the gene expression profiles of severe secondary pulmonary tuberculosis patients.Methods From May 2012 to October 2013,a total of 103 eligible patients with secondary pulmonary tuberculosis were recruited from Institution of Tuberculosis Research of PLA Hospital No.309.They were divided into severe secondary pulmonary tuberculosis (severe group) (n =57) and mild secondary pulmonary tuberculosis (mild group) (n =46) by the severity of disease.At the same time age-matched healthy controls (n =45) were selected from healthy subjects undergoing physical examination.Whole genome expression profiling was performed with Affymetrix Gene expression chips for 4 cases in severe group,3 in mild group and 5 in healthy group.Cluster and bioinformatics analysies were performed on differentially expressed genes in severe versus mild group.The remainders of three groups were 53,43 and 40 cases respectively used for verify the results of gene chip by real-time fluorescence quantitative PCR (RT-PCR).And 20 cases in severe group,20 in mild group and 8 in control group were used to verify the expression level of jun oncogene (JUN) on behalf of differential expressed genes.Analysis of variance and nun-parametric tests were used for statistic difference analysis among three groups.Results There were 406 differentially expressed genes for severe and mild groups.There were 264 down-regulated gene and 142 upregulated ones.The down-regulated genes were predominant.Cluster analysis show the similarity of gene expression profile in the same group.The result confirmed that the gene chip experiments were both repeatable and reliable.According to gene ontology,the differentially expressed genes were mainly involved in such biological processes as immune response,signal transduction,regulation of transcription (DNA-dependent),inflammatory response,antigen processing and presentation and chemotaxis,etc.Pathway analysis showed differentially expressed genes were involved in 22 pathways of immune response and inflammation.The major pathways included B cell receptor signaling,antigen processing and presentation,Toll-like receptor signaling,MAPK signaling and transforming growth factor-beta (TGF-β) signaling.Realtime fluorescence quantitative PCR (RT-PCR) analysis showed that the statistics of optical density for JUN was P ＜0.001 in severe versus mild group.It was down-regulated in severe group.And the expression of JUN was conformed with the result of gene expression chip.Conclusions The patients of severe group have a larger number of differential expressed genes versus those of mild group.And severe lung tissue damage in severe group may be correlated with differences in gene expression.
