塞莱昔布对幽门螺杆菌蛋白质组的影响  

Proteomic profiling of Helicobacter pylori treated with celecoxib

作　　者：高培培[1] 王蔚虹[1] 王静[1] 李江[1] 董欣红[1] 

机构地区：[1]北京大学第一医院消化内科,北京市100034

出　　处：《世界华人消化杂志》2011年第17期1785-1790,共6页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.30770981~~

摘　　要：目的:探讨选择性COX-2抑制剂塞莱昔布对幽门螺杆菌(H.pylori)蛋白质组的影响.方法:双向凝胶电泳技术(2-DE)分离塞莱昔布处理前后的H.pylori标准株26695全菌蛋白,银染法凝胶染色,图像扫描分析软件识别差异蛋白,基质辅助激光解吸电离飞行时间质谱(MALD-TOF-MS)、串联质谱(MALDI-TOF-MS/MS)分析差异蛋白;SYBRGreenⅠ实时定量PCR测定差异蛋白基因的表达变化.结果:塞莱昔布处理后,H.pylori标准株26695发现17个蛋白质斑点表达有差异.7个蛋白质斑点得到阳性鉴定,分别归为H.pylori的3种蛋白质:热休克蛋白60(HSP60),即groEL,延伸因子(EF-TU),谷氨酰转肽酶(GGT).与DMSO溶媒对照相比,塞莱昔布处理后的H.pyloriHSP60(groEL)、EF-TU、GGT表达下调.实时定量PCR测定显示,编码H.pyloriEF-TU、GGT基因的mRNA水平降低(0.07±0.06vs1.01±0.16;0.31±0.13vs0.98±0.01,均P<0.05),而编码H.pyloriHSP60(groEL)基因的mRNA水平增加(1.85±0.26vs1.07±0.27,P<0.05).结论:塞莱昔布通过下调H.pyloriEF-TU、GGTmRNA的表达减低其蛋白的表达;而其对HSP60蛋白质表达的下调作用不发生在mRNA转录水平.塞莱昔布可能通过调节H.pylori致病相关蛋白的表达影响其致病性.AIM： To perform a proteomic investigation of the effect of celecoxib on Helicobacter pylori （H.pylori）. METHODS： Total proteins of untreated and celecoxib-treated H.pylori 26695 were extracted and separated by 2-dimensionals polyacrylamide gel electrophoresis （2-DE）. Differential protein expression was detected using computer-assisted image analysis. Differential proteins were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF-MS） and matrix-assisted laser desorption/ionization time-of-flight-tandem mass spectrometry （MALDI-TOF-MS/MS）. The levels of mRNA expression were measured by realtime polymerase chain reaction.RESULTS： Seventeen differentially expressed spots were detected between untreated and celecoxib-treated H.pylori 26695. Seven spots were positively identified as three proteins： heat shock protein 60 （HSP60）, elongation factor TU （EF-TU） and gamma-glutamyltranspeptidase （GGT）. The protein expression of HSP60, GGT, and EF-TU, and mRNA expression of GGT and EF-TU were down-regulated （0.07 ± 0.06 vs 1.01 ± 0.16; 0.31 ± 0.13 vs 0.98 ± 0.01, both P 0.05）, while the mRNA expression of HSP60 was upregulated in the presence of celecoxib （1.85 ± 0.26 vs 1.07 ± 0.27, P 0.05）. CONCLUSION： Celecoxib could down-regulate the protein expression of HSP60, GGT and EFTU and mRNA expression of GGT and EF-TU in H.pylori; however, the mRNA expression of HSP60 was up-regulated. These results suggest that celecoxib might interfere with the pathogenicity of H.pylori.

关 键 词：幽门螺杆菌 塞莱昔布 蛋白质组 

分 类 号：R57[医药卫生—消化系统]