miR-142-3p靶向PERK/Bip信号通路对骨关节炎模型大鼠的干预作用  

Intervention effect of miR-142-3p targeting PERK/Bip signaling pathway in rats with osteoarthritis

作　　者：魏萌[1] 张英民[1] 康少英[1] 李建伟[1] 王振兴[1] 陈伟[1] WEI Meng;ZHANG Ying-min;KANG Shao-ying;LI Jian-wei;WANG Zhen-xing;CHEN Wei(Third Department of Orthopaedics,Handan Central Hospital,Handan Hebei 056001,China)

机构地区：[1]邯郸市中心医院骨三科,河北邯郸056001

出　　处：《局解手术学杂志》2022年第5期379-384,共6页Journal of Regional Anatomy and Operative Surgery

基　　金：邯郸市科学技术研究与发展计划(1823208046ZC)。

摘　　要：目的探究微小RNA-142-3p(miR-142-3p)靶向蛋白激酶R样内质网激酶(PERK)/免疫球蛋白结合蛋白(Bip)信号通路对骨关节炎模型大鼠的干预作用。方法选取60只SPF级雄性大鼠,随机挑选15只作为空白组,其余45只建立骨关节炎模型,最终39只建模成功,随机分为模型组、下调组、上调组,每组13只;建模成功后,空白组和模型组于膝部注射生理盐水,上调组和下调组分别于右膝关节腔内注射miR-142-3p mimic和miR-142-3p inhibitor。HE染色观察各组大鼠膝关节软骨组织病理学变化;观察并比较各组大鼠膝关节活动度、压痛阈值、热痛阈值;酶联免疫吸附试验(ELISA)检测血清白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)水平;实时荧光定量PCR检测转化生长因子-β(TGF-β)、Ⅰ型胶原α1(COL1α1)、金属蛋白酶组织抑制因子1(TIMP1)mRNA水平;Western blot检测PERK/Bip信号通路蛋白表达量。结果与模型组相比,下调组miR-142-3p表达量降低,上调组miR-142-3p表达量升高,差异有统计学意义(P<0.05),提示miR-142-3p转染成功。通过组织病理学观察发现,空白组和上调组大鼠膝关节软骨组织完整,模型组和下调组大鼠膝关节软骨退变,正常软骨结构丧失。与空白组比较,其他各组大鼠膝关节活动度较小,压痛阈值、热痛阈值较低,IL-1β、TNF-α、TGF-β、COL1α1、TIMP1 mRNA水平较高,PERK、Bip、Caspase-9蛋白表达较高,差异均有统计学意义(P<0.05);与模型组比较,下调组大鼠膝关节活动度较小,压痛阈值、热痛阈值较低,IL-1β、TNF-α、TGF-β、COL1α1、TIMP1 mRNA水平较高,PERK、Bip、Caspase-9蛋白表达较高,差异均有统计学意义(P<0.05);与模型组和下调组比较,上调组大鼠膝关节活动度较大,压痛阈值、热痛阈值较高,IL-1β、TNF-α、TGF-β、COL1α1、TIMP1 mRNA水平较低,PERK、Bip、Caspase-9蛋白表达较低,差异均有统计学意义(P<0.05)。结论上调骨关节炎大鼠miR-142-3p的表达可�Objective To investigate the intervention effect of microRNA-142-3p(miR-142-3p)targeting protein kinase R-like ER kinase(PERK)/immune globulin binding protein(Bip)signaling pathway in rats with osteoarthritis.Methods A total of 60 SPF male rats were selected,15 of which were randomly selected as the blank group,and the remaining 45 rats were established for osteoarthritis model.Finally,39 rats were successfully modeled and they were randomly divided into the model group,the down-regulated group and the up-regulated group,with 13 rats in each group.After successful modeling,the blank group and the model group were injected with normal saline in knees,and the up-regulated group and the down-regulated group were injected with miR-142-3p mimic and miR-142-3p inhibitor in the right knee cavity,respectively.Histopathological changes of knee cartilage in each group were observed by HE staining.Knee range of motion,tenderness threshold and thermal pain threshold in each group were observed and compared.Serum levels of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent assay(ELISA).The levels of transforming growth factor-β(TGF-β),typeⅠcollagenα1(COL1α1)and tissue inhibitor of metalloproteinase 1(TIMP1)mRNA were detected by real-time fluorescence quantitative PCR.The protein expression of PERK/Bip signaling pathway were detected by Western blot.Results Compared with the model group,the expression level of miR-142-3p was decreased in the down-regulated group and increased in the up-regulated group,with statistically significant differences(P<0.05),indicating that miR-142-3p was successfully transfected.Through histopathological observation,it was found that the knee cartilage tissues of rats in the blank group and the up-regulated group were intact,while the knee cartilage tissues of rats in the model group and the down-regulated group were degenerated and the normal cartilage structures were lost.Compared with the blank group,the other three groups had lower knee

关 键 词：PERK/Bip信号通路 微小RNA-142-3p 骨关节炎 炎症反应 滑膜纤维化 

分 类 号：R684.3[医药卫生—骨科学]