上调miR-155-5p对脊柱结核病模型小鼠的干预作用  

Intervention effects of upregulation of miR-155-5p in mice with spinal tuberculosis

作　　者：吴术其[1] WU Shuqi(Depatment of Thoracic Tuberculosis,Changsha Central Hospital,Hunan,Changsha 410007,China)

机构地区：[1]湖南省长沙市中心医院肺外结核科,410007

出　　处：《河北医药》2022年第15期2250-2253,2258,共5页Hebei Medical Journal

基　　金：湖南省卫生计生委科研计划课题项目(编号:B2016239)。

摘　　要：目的探究上调miR-155-5p对小鼠脊柱结核病小鼠的影响并探究破骨相关信号通路RANK/NF-κB信号通路及其磷酸化在此过程中发挥的作用。方法将40只C57BL/6小鼠分为对照组(n=10),模型组(n=10),NC组(n=10)和实验组(n=10),小鼠脊柱局部注射浓度为108 CFU/ml的H37Rv标准菌株悬液0.1 ml建立脊柱结核病模型,对照组注射等体积0.9%氯化钠溶液。NC组和实验组在模型建立后分别通过尾静脉注射miR-NC及miR-155-5p agomir,对照组和模型组注射等量0.9%氯化钠溶液,饲养8周后取脊柱标本送Micro-CT扫描检测计骨小梁数目和厚度,抗酒石酸酸性磷酸酶(Trap)染色观察破骨细胞数量,qPCR检测各组小鼠外周血中miR-155-5p表达水平,酶联免疫吸附剂测定法(Elisa)检测外周血白介素1-β(IL-1β),白介素-6(IL-6),白介素21(IL-17),肿瘤坏死因子α(TNF-α),Western blot检测结核病灶组织破骨相关蛋白核因子kB受体激活剂(RANK),核因子κB(NF-κB)及磷酸化核因子κB(p-NF-κB)蛋白的表达水平。结果qPCR结果表明与对照组比较,模型组和NC组小鼠miR-155-5p表达显著降低,实验组小鼠miR-155-5p显著升高(P<0.01),与对照组比较,模型组和NC组小鼠骨小梁数目和厚度均显著降低,实验组小鼠骨小梁数目和厚度均显著显著升高(P<0.01),Trap染色结果表明与对照组比较,模型组和NC组小鼠破骨细胞比例显著升高,而实验组破骨细胞显著降低。Elisa结果表明与对照组比较,模型组和NC组小鼠外周血炎性因子表达水平显著升高,实验组组外周血炎性因子表达显著降低(P<0.01)。Westernblot结果表明与对照组比较,模型组和NC组小鼠RANK、NF-κB及p-NF-κB蛋白表达具有升高趋势(P<0.01),实验组则有下降趋势(P<0.01)。结论脊柱结核病小鼠外周血miR-155-5p表达显著降低,上调miR-155-5p表达可以对小鼠脊柱结核具有保护作用,其机制可能与抑制炎性因子的释放及RANK/NF-κB信号通路的调控和磷酸化相�Objective To investigate the effects of upregulation of miR-155-5p on mice with spinal tuberculosis,and to explore the effects of osteoclast related signaling pathway RANK/NF-κB signaling pathway and its phosphorylation in this process.Methods Forty C57BL/6 mice were divided into control group,model group,NC group and experimental group,with 10 mice in each group.The animal models with pinal tuberculosis were established by injecting H37Rv standard strain suspension into the spinal column of mice,with the volume 0.1ml and concentration of 108 CFU/ml,and the mice in control group were given equal-volume 0.9%sodium chloride solution,and the mice in NC group and experimental group were injected with miR-NC and miR-155-5p agomir via tail vein after the animal models were established.After 8-week feeding,the number and thickness of trabecular bone were detected by Micro CT,and the number of osteoclasts was observed by tartrate resistant acid phosphatase(TRAP)staining,and the expression levels of miR-155-5p in peripheral blood were detected by qPCR,the expression levels of interleukin-1β(IL-1β),interleukin-6(IL-6),nterleukin-17(IL-17),tumor necrosis factorα(TNF-α)in peripheral blood were detected by enzyme-linked immunosorbent assay(ELISA),and the levels of osteoclast associated protein nuclear factor kB receptor activator(RANK),nuclear factor-κB(NF-κB)and phosphorylated nuclear factorκB(p-NF-κB)in tuberculosis tissue were detected by Western Blot.Results Compared with those in control group,the expression of levels miR-155-5p in model group and NC group were significantly decreased,however,the expression levels of miR-155-5p in experimental group were significantly increased(P<0.01).Compared with those in control group,the number and thickness of trabecular bone in model group and NC group were significantly decreased,and the number and thickness of trabecular bone in experimental group were significantly increased(P<0.01).Compared with that in in control group,the proportion of osteoclasts in model group and

关 键 词：miR-155-5p 脊柱结核病 炎性因子 RANK/NF-κB信号通路 保护作用 

分 类 号：R529.2[医药卫生—内科学]