微小RNA-7基因敲减对LPS诱导的脑部炎症的影响  

The effect of microRNA-7knock down on LPS-induced murine brain inflammation

作　　者：岳东旭[1,2] 赵娟娟[1,2] 胡琳 陈慧子 丁涛[1,2] 潘飞飞 陈超[1,2] 郭萌萌[1,2] 徐林[1,2] YUE Dong-xu;ZHAO Juan-juan;HU Lin;CHEN Hui-zi;DING Tao;PAN Fei-fei;CHEN Chao;GUO Meng-meng;XU Lin(Special Key Lab of Gene Detection and Treatment of Guizhou Province,Zunyi Medical University,Zunyi 563099,China;Department of Immunology,Talent Base of Biological Therapy of Guizhou Province,Zunyi Medical University,Zunyi 563099,China)

机构地区：[1]遵义医学院贵州省基因检测与治疗特色重点实验室,遵义563099 [2]遵义医学院免疫学教研室暨贵州省生物治疗人才基地,遵义563099

出　　处：《现代免疫学》2018年第5期366-371,共6页Current Immunology

基　　金：国家自然科学基金(31760258);贵州省高层次创新人才计划[黔科合人才(2016)4031号];遵义医学院优秀青年人才计划(15ZY-001)

摘　　要：为研究微小RNA-7(microRNA-7, miR-7)基因敲减(knock down, KD)对LPS诱导的脑部炎症的影响并探讨其意义,课题组用LPS腹腔注射(2.5mg/kg体质量)WT小鼠建立脑部炎性损伤模型;HE染色观察小鼠脑组织病理学变化,real-time PCR探针法检测脑组织中miR-7的表达变化;进一步,用LPS腹腔注射(2.5mg/kg体质量)WT小鼠和miR-7KD小鼠;12h后,HE染色观察小鼠脑组织病理学变化;real-time PCR检测脑组织中炎性细胞因子IL-6、TNF-α和TGF-β的mRNA变化水平;ELISA检测脑组织中炎性细胞因子IL-6、TNF-α和TGF-β的变化水平;Western blotting检测信号通路Akt和p-Akt以及炎性信号通路NF-κB和p-NF-κB的表达变化。结果显示小鼠炎性损伤模型脑组织中,HE示WT小鼠在注射LPS后,脑组织中炎性细胞浸润显著增多且miR-7的表达水平显著升高(P<0.01);HE结果示miR-7 KD小鼠脑组织中炎性细胞浸润较WT小鼠显著增多;real-time PCR结果示促炎细胞因子IL-6(P<0.01)和TNF-α(P<0.01)的表达水平均显著升高,而抑炎细胞因子TGF-β(P<0.01)的表达水平显著降低;ELISA结果显示,促炎细胞因子IL-6(P<0.01)和TNF-α(P<0.01)的表达水平均显著升高,而抑炎细胞因子TGF-β的表达水平显著降低(P<0.01); Western blotting结果示miR-7 KD模型小鼠脑组织Akt和p-Akt表达没有明显变化,NF-κB(P<0.05)和p-NF-κB(P<0.01)表达均明显增加。由此miR-7基因敲减显著促进了脑组织炎性损伤的发生,与NF-κB信号通路传递改变相关,本研究为后续深入探讨miR-7在脑部炎症相关疾病的发生机制中的作用提供了前期实验基础。We aimed to explore the effect of microRNA-7 knock down（KD）on LPS-induced brain inflammation in murine.Wild-type and miR-7 KD mice were intraperitoneally injected with LPS（2.5 mg/kg weight）to induce brain inflammation model.12 hours later,HE staining was used to observe the pathological changes in the brain tissues.The expressions of IL-6,TNF-α and TGF-β at RNA level were detected by real-time PCR and those at protein level were detected by ELISA.The changes of NF-κB and Akt signaling pathways were detected by Western blotting.Together,the miR-7 knock down in mice significantly enhanced the brain inflammation.The real-time PCR and Western blotting showed the increase in IL-6,TNF-α and the decrease in TGF-β in KD group（P 〈0.01）.Additionally,knock down of miR-7 results in changes in NF-κB signaling pathway while no changes were found for Akt pathway.In conclusion,miR-7 KD aggravates the inflammation in brain tissue and it affects NF-κB signaling pathway.These results would lay foundation for further study on the mechanism of miR-7 in the pathogenesis of brain inflammation-related diseases.

关 键 词：微小RNA-7 脑部炎症 NF-ΚB 细胞因子 

分 类 号：R394.33[医药卫生—医学遗传学]