赛培新增产品文献：大鼠IL-1β，IL-6，TNF-α ELISA试剂盒

发布时间：2022-04-08 10:53:33

miRNA-27a 对实验性肺结核大鼠免疫功能的调控作用及其机制

 韩 娜 1, 刘凡平 1, 田彦卿 1, 郑志清 1, 郎伟明 1, 王 倩 1, 刘亚涛 1, 朱建光 2 （1. 河北大学附属医院结核科，河北 保定 071000；2. 中国人民解放军陆军第八十二集团军医院 耳鼻喉科，河北 保定 071000） ［摘 要］ 目的目的：探讨 miRNA-27a 对实验性肺结核模型大鼠免疫功能的调控作用，阐明其可能的作 用机制。方法方法：经尾静脉注射结核杆菌悬液建立肺结核大鼠模型，将造模成功的大鼠随机分为模型 组、miR-27a 激动剂对照 （agomir-NC） 组和 miR-27a 激动剂 （agomir） 组，另取正常大鼠设为对照 组，每组 12 只。agomir-NC 组和 agomir 组大鼠经尾静脉注射 agomir-NC 或 miR-27a agomir 干预，每天 1 次，连续 5 d。3 周后，计算各组大鼠脾脏和胸腺指数，HE 染色观察各组大鼠肺组织病理形态表现， 流式细胞术检测各组大鼠外周血 T 淋巴细胞亚群百分率，酶联免疫吸附测定 （ELISA） 法检测各组大 鼠肺组织中白细胞介素 1β （IL-1β）、白细胞介素 6 （IL-6） 和肿瘤坏死因子 α （TNF-α） 水平，实时荧 光定量 PCR （RT-qPCR） 法检测各组大鼠肺组织中 miR-27a 表达水平，Western blotting 法检测各组大 鼠肺组织中 Toll样受体 4 （TLR4）、核因子 κB （NF-κB） 抑制蛋白 α （IκBα）、磷酸化 IκB （p-IκB） 和 NF-κB p65 蛋白表达水平。结果结果：与对照组比较，模型组大鼠肺组织损伤严重，脾脏及胸腺指数、外 周血中 CD4+T 淋巴细胞百分率和肺组织中 miR-27a 表达水平及 IκB 蛋白表达水平均明显降低 （P< 0. 05），外周血中 CD8+T 淋巴细胞百分率及肺组织中 IL-1β、IL-6、TNF-α 水平和 TLR4、p-IκB 及 NF-κB p65 蛋白表达水平均明显升高 （P<0. 05）；与模型组比较，agomir 组大鼠肺组织病理形态表现 明显改善，脾脏和胸腺指数、外周血中 CD4+T 淋巴细胞百分率、肺组织中 miR-27a 表达水平及 IκB 蛋白表达水平均明显升高 （P<0. 05），外周血中 CD8+T 淋巴细胞百分率及肺组织中 IL-1β、IL-6、 TNF-α 水平和 TLR4、p-IκB、NF-κB p65 蛋白表达水平均明显降低 （P<0. 05），agomir-NC 组大鼠上 述各指标差异均无统计学意义 （P>0. 05）。结论结论：过表达 miR-27a 可能通过抑制 TLR4/NF-κB 信号通 路活化，减少炎症因子释放，调节机体免疫反应，从而改善肺结核大鼠肺损伤。 ［关键词］］ 微小 RNA；miRNA-27a；肺结核；Toll样受体 4/核因子 κB 信号通路；免疫功能；炎症 ［中图分类号］ R-332; R521 ［文献标志码］ A 

Regulatory effect of miRNA-27a on immune function in experimental pulmonary tuberculosis rats and its mechanism

HAN Na1 , LIU Fanping1 , TIAN Yanqing1 , ZHENG Zhiqing1 , LANG Weiming1 , WANG Qian1, LIU Yatao1 , ZHU Jianguang2 （1. Department of Tuberculosis，Affiliated Hospital，Hebei University，Baoding 071000，China； 2. Department of Otolaryngology，No.82 Military Hospital of PLA，Baoding 071000，China） ABSTRACT Objective：To investigate the regulatory effect of miRNA-27a on the immune function of the experimental pulmonary tuberculosis model rats，and to clarify its possible mechanism. Methods：The rat [文章编号] 1671-587X(2022)01-0104-07 [收稿日期] 2021-05-24 [基金项目] 河北省卫健委医学科学研究计划项目 （20190937） [作者简介] 韩 娜 （1981－），女，河北省保定市人，副主任医师，医学硕士，主要从事肺结核基础和临床方面的研究。 [通信作者] 朱建光，副主任医师 （E-mail：zxfj6098@163.com） 104韩 娜，等 . miRNA-27a 对实验性肺结核大鼠免疫功能的调控作用及其机制 models of pulmonary tuberculosis were established by injecting Mycobacterium tuberculosis suspension through tail vein，after successful modeling，the model rats were randomly divided into model group，miR- 27a agomir control（agomir-NC）group and miR-27a agomir（agomir）group，and another normal rats were used as control group；there were 12 mice in each groups. The rats in agomir-NC group or miR-27a agomir group were injected with agomir-NC or miR-27a agomir via tail vein once a day for 5 consecutive days. Three weeks later，the spleen and thymus indexes of the rats in various groups were calculated，the pathomorphology of lung tissue of the rats in various groups were observed by HE staining；the percentages of T lymphocyte subsets in peripheral blood of the rats in various groups were detected by flow cytometry； the levels of interleukin-1β（IL-1β），interleukin-6（IL-6）and tumor necrosis factor- α（TNF- α）in lung tissue of the rats in various groups were determined by enzyme linked immunosorbent assay（ELISA）；the expression levels of miR-27a in lung tissue of the rats in various groups were detected by real-time fluorescence quantificative PCR （RT-qPCR） method； the expression levels of Toll-like receptor 4 （TLR4），nuclear factor kappa-B（NF-κB）inhibitor α（IκBα），phosporylated-IκB（p-IκB）and NF-κB p65 in lung tissue were detected by Western blotting method. Results：Compared with control group，the lung injury of rats in model group was serious，the spleen and thymus indexes，the percentages of CD4+T cells in peripheral blood，the expression level of miR-27a and the expression level of IκB protein in lung tissue were significantly decreased（P<0. 05）；the percentages of CD8+T cells in peripheral blood，the levels of IL-1β，IL-6，TNF-α and the expression levels of TLR4，p-IκB and NF-κB p65 proteins in lung tissue were significantly increased（P<0. 05）. Compared with model group，the pathomorphology of lung tissue of the rats in agomir group was significantly improved，the spleen and thymus indexes，the percentages of CD4+ T cells in peripheral blood，the expression level of miR-27a and the expression level of IκB protein in lung tissue were significantly increased（P<0. 05）；the percentage of CD8+T cells in peripheral blood，the levels of IL-1β，IL-6，TNF- α and the expression levels of TLR4，p-IκB and NF- κB p65 proteins in lung tissue of the rats were significantly decreased（P<0. 05）；there were no statistically significant differences in the above indexes of the rats in agomir-NC group（P>0. 05）. Conclusion：Overexpression of miR-27a may improve the lung injury in the pulmonary tuberculosis rats by inhibiting activation of TLR4/NF- κB signaling pathway，reducing the release of inflammatory factors and regulating the body's immune response. KEYWORDS MiRNA-27a； Pulmonary tuberculosis； Toll-like receptor 4/nuclear factor kappa-B signaling pathway；Immune function；Inflammation

miRNA-27a对实验性...免疫功能的调控作用及其机制_韩娜 (1).pdf