人β-防御素在空肠弯曲菌感染中的表达与调控  

Expression and regulation of human beta-defensin in Campylobacter jejuni infection

作　　者：吴琼[1] 陈洪友[2] 屠丽红[2] 宋元君[2] 张雯霞[2] 陈敏[2] 

机构地区：[1]上海交通大学附属第六人民医院,上海200233 [2]上海市疾病预防控制中心,上海200336

出　　处：《检验医学》2016年第8期662-666,共5页Laboratory Medicine

基　　金：上海市自然科学基金资助项目(11ZR1434200)

摘　　要：目的调查2012至2013年上海地区腹泻症候群调查研究及病原谱监测中空肠弯曲菌的感染现状,探讨在空肠弯曲菌引起的肠道感染中人β-防御素(hBD)的抗菌作用及其免疫调节功能。方法收集2012至2013年上海地区腹泻症候群调查研究及病原谱监测中分离到的空肠弯曲菌,分析其感染现状;采用琼脂稀释法检测空肠弯曲菌对庆大霉素(GEN)、四环素(TET)、阿奇霉素(AZI)、红霉素(ERY)、克林霉素(CLI)、环丙沙星(CIP)、萘啶酸(NAL)的最低抑菌浓度(MIC);采用十二烷基硫酸钠-脉冲场凝胶电泳(SDS-PFGE)分析其分子流行趋势;空肠弯曲菌标准菌株感染人类肠上皮细胞系Caco-2,采用荧光定量聚合酶链反应(PCR)检测肠上皮细胞hBD-1、hBD-2、hBD-3、TLR2和TLR4基因转录情况;采用蛋白免疫印迹法检测hBD-1、hBD-2、hBD-3、TLR2、TLR4蛋白表达情况。结果 2012至2013年上海地区腹泻患者空肠弯曲菌分离率为1.6%。37株空肠弯曲菌对TET和NAL的耐药性极高,分别达95.7%和97.8%。PFGE分为9个型别。hBD-1、hBD-2、hBD-3在Caco-2细胞感染空肠弯曲菌6 h时转录水平最高,而其蛋白表达量则在24 h时最高。TLR2在感染24 h的基因转录水平较0、6、10 h有显著升高,但TLR4则在感染6 h时基因转录水平最高,TLR2、TLR4蛋白表达与基因转录水平相一致。结论 hBD在空肠弯曲菌感染中起着一定的防御作用。Objective To investigate the infection status of Campylobacter jejuni isolated from diarrhea syndrome and pathogenic monitoring study in Shanghai from 2012 to 2013. To study the immune regulation of human beta-defensin （hBD） in gastrointestinal infection caused by Campylobacter jejuni. Methods Campylobacter jejuni were isolated from diarrhea syndrome and pathogenic monitoring study in Shanghai from 2012 to 2013. Minimal inhibitory concentrations（MIC） to 7 antibiotics [gentamycin（GEN）, tetracycline（TET）,azithromycin（AZI）,erythromycin（ERY）,clindamycin（CLI）,ciprofloxacin （CIP） and nalidixic acid（NAL）] were determined by agar dilution method. Sodium dodecyl sulfate-pulsed-field gel electrophoresis （SDS-PFGE） was carried out for analyzing molecular epidemic trend. Human intestinal epithelial cell line Caco-2 was infected with Campylobacter jejuni. The transcription levels of hBD-1, hBD-2,hBD-3,TLR2 and TLR4 genes were determined by fluorescence quantitation polymerase chain reaction （PCR）,and the expressions of hBD-1,hBD-2,hBD-3,TLR2 and TLR4 were determined by western blotting. Results The isolation rate of Campylobacter jejuni isolated from diarrhea syndrome and pathogenic monitoring study in Shanghai from 2012 to 2013 was 1.6%. The drug resistance rates to TET and NAL were high （95.7% and 97.8%）. There were 9 PFGE patterns. After Campylobacter jejuni infected human intestinalepithelial cell line Caco-2,the transcription levels of hBD-1,hBD-2 and hBD-3 genes reached the highest at 6 h, and the expressions of hBD-1,hBD-2 and hBD-3 reached the highest at 24 h. The transcription level of TLR2 at 24 h was higher than those at 0,6 and 10 h. However,the transcription level of TLR4 reached the highest at 6 h. The expressions of TLR2 and TLR4 were the same as their transcription levels. Conclusions hBD plays a role in defense of Campylobacter jejuni infection.

关 键 词：人β-防御素 空肠弯曲菌 免疫防御 

分 类 号：R446.5[医药卫生—诊断学]