miR-129靶向HMGB1对大鼠急性脊髓损伤后炎症反应的影响  

Effect of miR-129 targeting HMGB1 on inflammatory response after acute spinal cord injury in rats

作　　者：黄炜[1] 贺媛 张卫涛 HUANG Wei;HE Yuan;ZHANG Wei-tao(Department of Rehabilitation,Baoji Vocational and Technical College,Shaanxi,721000,China;Department of Rehabilitation,Xi’an Traditional Chinese Medicine Encephalopathy Hospital,Shaanxi,710032,China;Department of Orthopedics,Xi’an Honghui Hospital,Shaanxi,710054,China)

机构地区：[1]宝鸡职业技术学院康复教研室,陕西宝鸡721000 [2]西安中医脑病医院康复治疗科,陕西西安710032 [3]西安市红会医院骨科,陕西西安710054

出　　处：《颈腰痛杂志》2023年第3期305-310,共6页The Journal of Cervicodynia and Lumbodynia

基　　金：国家社会科学基金重点项目(编号:17AYY008)。

摘　　要：目的探讨微小RNA-129(mini RNA-129,miR-129)对大鼠急性脊髓损伤(acute spinal cord injury,ASCI)后炎症反应的影响及其机制。方法实验分为正常组、假手术组、模型组、miR-NC(阴性)组、miR-129 agomir(激动剂)组,每组12只。对各组大鼠进行BBB脊髓运动功能评分;通过H-E染色进行脊髓组织形态学观察;通过免疫荧光染色观察A1型星形胶质细胞;通过实时荧光定量聚合酶链式反应(qRT-PCR)检测miR-129和靶基因高迁移率族蛋白B1(high mobility group box 1,HMGB1)mRNA的表达;通过蛋白质免疫印迹技术(western blot,WB)检测HMGB1蛋白表达;通过酶联免疫吸附试验检测肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)和IL-6表达水平;通过双荧光素酶法检测miR-129和HMGB1靶向关系,实验分为四组:pMIR-HMGB1-WT+miR-129 mimics组、pMIR-HMGB1-WT+miR-NC组、pMIR-HMGB1-MUT+miR-129 mimics组、pMIR-HMGB1-MUT+miR-NC组。结果与正常组相比,假手术组miR-129表达、HMGB1mRNA和蛋白表达、TNF-α、IL-1β、IL-6水平以及A1型星形胶质细胞水平差异无统计学意义(P>0.05);模型组miR-129的表达显著降低,HMGB1mRNA和蛋白表达、TNF-α、IL-1β、IL-6水平以及A1型星形胶质细胞水平均显著升高(P<0.05)。与模型组相比,miR-129NC组上述指标差异无统计学意义(P>0.05);miR-129 agomir组miR-129的表达显著升高,HMGB1mRNA和蛋白表达、TNF-α、IL-1β和IL-6水平以及A1型星形胶质细胞水平均显著降低(P<0.05)。与pMIR-HMGB1-WT+miR-NC组相比,pMIR-HMGB1-WT+miR-129组荧光素酶活性显著降低(P<0.05);pMIR-HMGB1-MUT+miR-129组和pMIR-HMGB1-MUT+miR-NC组之间无明显差异(P>0.05)。结论miR-129可靶向调控HMGB1,下调炎性因子TNF-α、IL-1β和IL-6水平,减轻大鼠ASCI发生后的炎症反应。Objective To investigate the effect of mini RNA-129(miR-129)on inflammatory response after acute spinal cord injury(ASCI)in rats and its mechanism.Methods The experiment was divided into normal group,sham operation group,model group,miR-NC(negative)group and miR-129 agomir(agonist)group,with 12 rats in each group.The evaluation of Basso-Beattie-Bresnahan(BBB)score was performed;hematoxylin eosin staining was used to observe the histomorphology of spinal cord;immunofluorescence staining was used to observe type A1 astrocytes;real-time fluorescent quantitative polymerase chain reaction(qRTPCR)was used to detect the expression of miR-129 and high mobility group box 1(HMGB1)mRNA;Western blot(WB)was applied to detect the expression of HMGB1 protein;enzyme-linked immunosorbent assay(ELISA)was employed to detect the expression levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and IL-6;double luciferase assay was used to detect the targeting relationship between miR-129 and HMGB1.The experiment was divided into four groups:pMIR-HMGB1-WT+miR-129 mimics group,pMIR-HMGB1-WT+miR-NC group,pMIR-HMGB1-MUT+miR-129 mimics group,and pMIR-HMGB1-MUT+miR-NC group.Results Compared with those in the normal group,there were no significant differences in the expression of miR-129,the expression of HMGB1 mRNA and protein,the levels of TNF-α,IL-1βand IL-6,and the level of A1 type astrocytes in the sham operation group(P>0.05);in the model group,the expression of miR-129 was significantly lower,the expression of HMGB1 mRNA and protein,the levels of TNF-α,IL-1βand IL-6,and the level of A1 type astrocytes were significantly higher(P<0.05).Compared with those in the model group,there was no significant difference in the above indexes in miR-129-NC group(P>0.05);in the miR-129 agomir group,the expression of miR-129 was significantly higher,the expression of HMGB1 mRNA and protein,the levels of TNF-α,IL-1βand IL-6,and the level of A1 type astrocytes were significantly lower(P<0.05).Compared with pMIR-HMGB1-WT+miR-NC group,t

关 键 词：miR-129 HMGB1 急性脊髓损伤 炎症反应 星形胶质细胞 

分 类 号：R744[医药卫生—神经病学与精神病学]