摘要
目的探讨潜伏结核感染和活动性缔结核感染对患者外周血CD4^＋T细胞内miR-29家族及其靶基因IFN-γ表达的影响。方法于2012年3-12月，在山东省潍坊市某两家医院选取调查对象并分为3组：活动性肺结核组（30例）、潜伏结核感染（LTBI）组（25例）和健康对照组（30名）。分离纯化3组受试者外周血中的CD4^＋T细胞；州核酸杂交与荧光定量PCR检测CD4^＋T细胞内miR-29a、miR-29b和miR-29c的表达；用定量PCR检测miR-29靶基因IFN-γ的表达；用TargetScan与PicTar联合预测miR-29家族的靶基因；用David数据库和Cytoscape软件对miR-29家族的预测靶基因进行皋因本体论（GO）和京都基因和基因组百科仝书（KEGG）生物通路富集分析。结果miR-29a、miR-29b和miR-29c在活动性肺结核组、LTBI组和健康对照组CD4^＋T细胞中的表达水平均不同（P〈0．05）：miR-29b和miR-29c在活动性肺结核组的表达（561．63±65．36，281．85±42．78）高于健康对照组（260．74±38．69，128．21±19．98），低于LTBI组（2030．29±321．68，620．93±79．14）；miR-29a在对照组的表达水平（913．95±104．73）高于活动性结核组（323．37±54．38），低于LTBI组（4782．13±567．81）。靶基凶IFN-γ在3组之间的表达水平亦不同（P〈0．05）：LTBI组（0．45±0．09）低于健康埘照组（1．00），而高于活动性结核组（0．11±0．03）。miRNA-29家族的预测靶基冈的GO功能富集于细胞外基质结构成分、转录调节活性等分子功能方面；在KEGG的通路数据库中，miR-29家族预测靶基因集合屁著富集于局部黏附信号通路、调节细胞骨架与mTOR信号通路等方面。结论LTBI和活动性结核感染明显增加了患者外周血CD4^＋T细胞内miR-29家族的表达，降低了其靶基因IFN-γ的表达，从而影响了信号通路等生物学过程。
Objective To investigate the effect of latent and active pulmonary tuberculosis （TB） on expression of miR-29 family and target gene IFN-γ in CD4^±T cells. Methods Subjects from two hospitals of Weifang were enrolled from March 2012 to December 2012 and divided into three groups： active TB group （30 cases） ,latent tuberculosis infection （LTBI） group （ 25 cases） and healthy control group （ 30 cases）. CD4^± T cells in blood were collected fi＇oin the three groups. Levels of miR-29a,miR-29b and miR-29c were measured by nucleic acid hybridization and RT-qPCR. Expression of IFN-γ was analyzed by RT-qPCR. Target genes of miR-29 family were predicted with both TargetScan and PicTar. GO annotation and pathway ovelxepresentation were further analyzed with David database and Cytoscape. Results Levels of miR-29a, miR-29b and miR-29c showed significant differences among the three groups（ P 〈 0. 05 ） ： levels of miR-29b and miR-29c in the active TB group （ 561.63 _± 65.36,281.85 ± 42. 78 ） were higher than the healthy controls（260. 74 ±38.69,128.21 ± 19.98） ,but lower than the LTBI group（2030. 29 ± 321.68,620. 93 ±79. 14） ; expression of miR-29a in the healthy control group（913.95 ± 104. 73） were higher than the active TB group（323.37±54. 38 ）, but lower than the LTBI group （4782. 13 ± 567.81 ）. Level of IFN-3, showed significant differences among the three groups （ P 〈 0. 05 ） ： level of IFN-3, in the LTBI group （ 0. 45 _± 0. 09 ） were lower than the healthy controls （ 1.00）, but higher than the active TB group （0. 11 -4- 0. 03 ）. The target genes of miR-29 family mainly existed in molecular function such as extraeellular matrix structural constituent and transcription regulator activity. In KEGG pathway,the gene set mostly existed in signaling pathway such as Focal adhesion, ECM-receptor interaction and mTOR signaling pathway. Conclusion The expression of miR-29 family was increased and target gene IFN-γ in CD4^± T cells was decreased by latent and active pulmonary TB,which might play important role in alteration of signal pathway.
出处
《中华预防医学杂志》
CAS
CSCD
北大核心
2013年第7期632-636,共5页
Chinese Journal of Preventive Medicine
基金
国家自然科学基金（81100006）
山东省自然科学基金（ZR2010HM073）
山东省高等学校科技计划项目（J09LF20）
作者简介
通信作者：付玉荣，Email：yifuyurong@163．com
