摘要
目的探讨丹参酮ⅡA(TanⅡA)通过介导Yes激酶相关蛋白(Yes-associated protein,YAP)、核因子-κB受体活化因子配基(receptor activator of nuclear factor-κB ligand,RANKL)/核因子κB受体活化因子(eceptor activator of nuclear factor-κB,RANK)/骨保护蛋白(osteoprotegerin,OPG)调节骨关节炎小鼠骨代谢的作用机制。方法建立骨关节炎小鼠模型,将60只小鼠随机分成假手术组、模型组、TanⅡA低剂量组和TanⅡA高剂量组,每组15只,造模成功后灌胃给药,连续4周。HE和番红O固绿染色观察软骨组织病理损伤并进行Mankin评分。酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)检测血清骨碱性磷酸酶(bone alkaline phosphatase,BALP)、骨钙素(osteocalcin,OC)、Ⅰ型胶原交联羧基末端肽(C-telopeptide of typeⅠcollagen,CTX)、白细胞介素(interleukin,IL)-1β、IL-6、IL-8和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)。蛋白质印迹法(Western blotting)检测基质金属蛋白酶(matrix metalloproteinases,MMPs)、YAP、RANK、RANKL和OPG蛋白。结果TanⅡA可改善小鼠软骨组织病理变化并降低Mankin评分。与假手术组比较,模型组BALP、OC水平下降,CTX、TNF-α、IL-6、IL-1β、IL-8、MMP1、MMP3和MMP13水平升高(P<0.05)。与模型组比较,TanⅡA低剂量组、TanⅡA高剂量组BALP、OC水平升高,CTX、TNF-α、IL-6、IL-1β、IL-8、MMP1、MMP3和MMP13水平降低(P<0.05)。与假手术组比较,模型组小鼠软骨组织中YAP、OPG和RANK蛋白水平下降,RANKL蛋白水平升高(P<0.05);与模型组比较,TanⅡA 2组小鼠软骨组织中YAP、OPG和RANK蛋白水平上升,RANKL蛋白水平下降(P<0.05)。结论TanⅡA可能通过介导YAP、RANK/RANKL/OPG信号通路调控骨关节炎。
Objective To explore the action mechanism of tanshinoneⅡA(TanⅡA)on regulating bone metabolism in osteoarthritis mice by mediating Yes-associated protein(YAP)and receptor activator of nuclear factor-κB ligand(RANKL)/receptor activator of nuclear factor-κB(RANK)/osteoprotegerin(OPG).Methods The osteoarthritis models of mice were prepared.A total of 60 mice were randomly divided into sham operation group,model group,low-dose and high-dose TanⅡA groups,15 cases in each group.After successful modeling,they were given intragastric administration for 4 weeks.The pathological damage of cartilage tissues was observed by HE and Safranin O fast green staining,and Mankin scoring was conducted.The levels of serum bone alkaline phosphatase(BALP),osteocalcin(OC),C-telopeptide of typeⅠcollagen(CTX),interleukin(IL)-1β,IL-6,IL-8 and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent assay(ELISA).The expressions of matrix metalloproteinases(MMPs),YAP,RANK,RANKL and OPG were detected by Western blotting.Results TanⅡA could improve pathological changes of cartilage tissues and decrease Mankin score.Compared with sham operation group,the levels of BALP and OC were decreased in model group,while the levels of CTX,TNF-α,IL-6,IL-1β,IL-8,MMP1,MMP3 and MMP13 were increased(P<0.05).Compared with model group,the levels of BALP and OC were increased in low-dose and high-dose TanⅡA groups,while the levels of CTX,TNF-α,IL-6,IL-1β,IL-8,MMP1,MMP3 and MMP13 were decreased(P<0.05).Compared with sham operation group,the levels of YAP,OPG and RANK in cartilage tissues were decreased,while the RANKL level was increased in model group(P<0.05).Compared with model group,the levels of YAP,OPG and RANK in cartilage tissues were increased,while the RANKL level was decreased in low-dose and high-dose TanⅡA groups(P<0.05).Conclusion TanⅡA may regulate osteoarthritis by mediating YAP and RANK/RANKL/OPG signaling pathways.
作者
张超
周迎锋
路坦
赵红星
耿晓林
陶金刚
徐海斌
ZHANG Chao;ZHOU Yingfeng;LU Tan;ZHAO Hongxing;GENG Xiaolin;TAO Jingang;XU Haibin(The First Ward,Department of Orthopedic Surgery,the First Affiliated Hospital of Xinxiang Medical College,Weihui 453100,China)
出处
《西北药学杂志》
CAS
2024年第2期74-80,共7页
Northwest Pharmaceutical Journal
基金
2019年度新乡医学院第一附属医院青年培育基金项目(编号:QN-2019-B04)。
