目的 探讨丹参酮Ⅱ A(Tan Ⅱ A)通过介导Yes激酶相关蛋白(Yes-associated protein，YAP)、核因子-KB受体活化因子配基(receptor activator of nuclear factor-κB ligand，RANKL)/核因子 KB 受体活化因子(eceptor activator of nuclear factor-KB，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、Tan ⅡA低剂量组和Tan Ⅱ A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 type Ⅰ collagen，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 Tan ⅡA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，Tan ⅡA低剂量组、Tan ⅡA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，Tan Ⅱ A 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 Tan ⅡA可能通过介导YAP、RANK/RANKL/OPG信号通路调控骨关节炎。
Action mechanism of tanshinone Ⅱ A on regulating bone metabolism in osteoar-thritis mice
Objective To explore the action mechanism of tanshinone Ⅱ A(Tan Ⅱ A)on regulating bone metabolism in osteoar-thritis mice by mediating Yes-associated protein(YAP)and receptor activator of nuclear factor-κB ligand(RANKL)/receptor activator of nuclear factor-KB(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 Tan Ⅱ A 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 type Ⅰ collagen(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 Tan Ⅱ A 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 Tan Ⅱ A 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 Tan Ⅱ A groups(P＜0.05).Conclusion Tan Ⅱ A may regulate osteoarthritis by mediating YAP and RANK/RANKL/OPG signaling pathways.
tanshinone ⅡAosteoarthritisYes-associated proteinreceptor activator of nuclear factor-κB ligandreceptor activator of nuclear factor-κBosteoprotegerin
NETL
NSTL
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