摘要
目的 探讨电热针通过调控Wnt/PPARγ信号通路对膝骨性关节炎大鼠滑膜炎性及软骨细胞凋亡的影响。方法 选取SPF级SD雄性大鼠40只,随机分为正常(NOR)组,模型(MOD)组,电热针(EHN)组,PPARγ激动剂(AGO)组,每组10只。对MOD组、EHN组、AGO组采用膝关节腔注射Ⅱ型胶原酶法建立膝骨性关节炎模型,NOR组不建立该模型,建模成功后,对EHN组行电热针治疗,对AGO组腹腔注射30 mg/kgWnt/PPARγ信号通路激动剂吡格列酮,NOR组、MOD组同期灌胃同体积0.9%氯化钠溶液,记录膝关节活动度及肿胀度,HE染色检测滑膜组织病理形态,酶联免疫吸附法检测滑膜炎症,TUNEL法检测软骨细胞凋亡,免疫印迹法检测Wnt/PPARγ信号通路相关蛋白表达。结果 与NOR组相比,MOD组膝关节活动度显著降低(P<0.05),膝关节肿胀度显著升高(P<0.05),与MOD组比较,EHN组、AGO组膝关节活动度显著升高(P<0.05),膝关节肿胀度显著降低(P<0.05);NOR组滑膜组织正常,滑膜细胞排列整齐,无血管翳形成,也未见明显炎症细胞浸润,MOD组出现滑膜细胞增生,有血管翳形成及明显炎症细胞浸润情况,与MOD组比较,EHN组、AGO组病理结构明显改善;与NOR组相比,MOD组大鼠滑膜组织TGF-β1、IL-1α、IL-21显著升高(P<0.05),与MOD组比较,EHN组、AGO组大鼠滑膜组织TGF-β1、IL-1α、IL-21显著降低(P<0.05);与NOR组相比,MOD组大鼠软骨细胞凋亡率显著升高(P<0.05),与MOD组比较,EHN组、AGO组大鼠软骨细胞凋亡率显著降低(P<0.05);与NOR组相比,MOD组大鼠膝关节组织Wnt4、β-catenin蛋白表达显著升高(P<0.05),PPARγ蛋白表达显著降低(P<0.05),与MOD组比较,EHN组、AGO组大鼠膝关节组织Wnt4、β-catenin蛋白表达显著降低(P<0.05),PPARγ蛋白表达显著升高(P<0.05)。结论 电热针可有效抑制膝骨性关节炎大鼠滑膜炎症及软骨细胞凋亡,其作用机制可能与调控Wnt/PPARγ信号通路有关。
Objective To investigate the effects of electrothermic needle on synovial inflammation and chondrocyte apoptosis in rats with knee osteoarthritis by regulating the Wnt/PPARγsignaling pathway.Methods Forty Sprague-Dawley(SD)male rats in the specific pathogen free(SPF)grade were randomly divided into control(NOR)group,model(MOD)group,electrothermic needle(EHN)group and PPARγagonist(AGO)group,with 10 rats in each group.Knee osteoarthritis model was established by injections of typeⅡcollagenase into the knee space of rats in MOD,EHN and AGO groups,and rats in the NORgroup were treated with blank control.After the modeling was successfully created,rats in the EHN group were treated with electrothermic needle,and those in the AGO group were intrabitoneally injected with 30mg/kg Wnt/PPARγsignaling pathway agonist pioglitazone.Rats in the NORgroup and MOD group were given the same volume of normal saline via oral gavage.Knee joint activity and swelling were recorded,and the pathological morphology of the synovial tissue was detected by hematoxylin and eosin(H&E)staining.Synovial inflammation was detected by enzyme-linked immunosorbent assay(ELISA),and chondrocyte apoptosis was detected by the terminal deoxynucleotidyl transferase dUTP nick-end labeling(TUNEL)assay.Protein levels of relevant genes in the Wnt/PPARγsignaling pathway were detected by Western blot.Results Compared with those of NORgroup,rats in the MOD group presented significantly decreased knee joint activity and increased knee joint swelling(P<0.05).Compared with those of MOD group,rats in the EHN group and AGO group presented significantly increased knee joint activity and decreased knee joint swelling(P<0.05).In the NORgroup,rats had normal synovial tissue with neat arrangement of synovial cells,no pannus formation and no obvious inflammatory cell infiltration.Rats in the MOD group had synovial cell proliferation,pannus formation and obvious inflammatory cell infiltration.Compared with those of MOD group,the pathological structures of key joint in rats of EHN group and AGO group were significantly alleviated.Compared with those of NORgroup,relative levels of TGF-β1,IL-1αand IL-21 in synovial tissue of rats in MOD group were significantly elevated(P<0.05),while TGF-β1,IL-1αand IL-21 in synovial tissue of rats in EHN and AGO groups were significantly reduced compared with those of MOD group(P<0.05).Compared with that of NORgroup,the apoptotic rate of chondrocytes in the MOD group was significantly elevated(P<0.05).Compared with that of MOD group,the apoptotic rate of chondrocytes in EHN group and AGO group was significantly reduced(P<0.05).Compared with those of NORgroup,rats in the MOD group had significantly upregulated Wnt4 andβ-catenin,and downregulated PPARγin knee tissue(P<0.05).Downregulated Wnt4 andβ-catenin,and upregulated PPARγwere observed in knee tissue of rats in EHN group and AGO group(P<0.05).Conclusion Electrothermic needle can effectively inhibit synovial inflammation and chondrocyte apoptosis in rats with knee osteoarthritis by regulating the Wnt/PPARγsignaling pathway.
作者
杨坤
胡锋
闵慧
梅求安
许明军
YANG Kun;HU Feng;MIN Hui(Department of Orthopedic Rehabilitation,Shiyan Taihe Hospital(Affiliated Hospital of Hubei University of Medicine),Hubei,Shiyan 442000,China;不详)
出处
《河北医药》
2025年第8期1253-1257,共5页
Hebei Medical Journal
基金
湖北省中医药管理局中医药科研项目(编号:ZY2023F072)。
