橄榄苦苷调节Hippo-YAP/TAZ信号通路对膝骨关节炎大鼠滑膜炎症的影响

Effect of oleuroside on synovial inflammation in knee osteoarthritis rats by regulating the Hippo-YAP/TAZ signaling pathway

目的探讨橄榄苦苷(OL)通过调节Yes相关蛋白(YAP)/转录共激活因子PDZ结合基序(TAZ)信号通路对膝骨关节炎(KOA)大鼠滑膜炎症的影响。方法采用改良的Hulth法构建KOA大鼠模型,并随机分为Model组、低剂量OL组(20 mg/kg)、高剂量OL组(40 mg/kg)、高剂量OL+VT103组(40 mg/kg+10 mg/kg YAP/TAZ抑制剂),每组12只,另取12只大鼠作为正常对照组(NC组)。观察各组大鼠的情况,Lequesne MG评分对各组大鼠行为学进行评估;ELISA试剂盒检测大鼠血清中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)的表达;苏木精-伊红(HE)染色检测大鼠滑膜组织的病理损伤;qRT-PCR法检测滑膜组织中YAP和TAZ mRNA的表达;Western blot检测大鼠滑膜组织中YAP、磷酸化YAP(p-YAP)、磷酸化TAZ(p-TAZ)、TAZ、B细胞淋巴瘤/白血病-2相关X蛋白(Bax)蛋白表达。结果与NC组比较,Model组大鼠Lequesne MG评分升高、血清中TNF-α和IL-6的表达增加、YAP和TAZ mRNA表达增加、p-YAP/YAP、p-TAZ/TAZ、Bax蛋白表达升高(P<0.05);与Model组比较,低剂量、高剂量OL组大鼠Lequesne MG评分降低、血清中TNF-α和IL-6的表达减少、YAP和TAZ mRNA表达减少、p-YAP/YAP、p-TAZ/TAZ、Bax蛋白表达降低(P<0.05);VT103可明显减弱OL对KOA大鼠滑膜组织的保护作用(P<0.05)。结论OL可能通过激活YAP/TAZ信号通路减轻膝骨关节炎大鼠滑膜炎症。

Objective To investigate the effect of oleuroside(OL)on synovial inflammation in knee osteoarthritis(KOA)rats by regulating the Yes-associated protein(YAP)/transcriptional co-activator with PDZ-binding motif(TAZ)signaling pathway.Methods A KOA rat model was constructed using the improved Hulth's method and randomly grouped into a Model group,a low-dose OL group(20 mg/kg),a high-dose OL group(40 mg/kg),and a high-dose OL+VT103 group(40 mg/kg+10 mg/kg YAP/TAZ inhibitor),with 12 rats in each group,another 12 rats were taken as the normal control group(NC group).The condition of rats in each group was observed,and the behavior of rats in each group was evaluated with the Lequesne MG score;ELISA kit was applied to detect the expression of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in serum;hematoxylin-eosin(HE)staining was applied to detect pathological damage in synovial tissue;qRT-PCR method was applied to detect the expression of YAP and TAZ mRNA in synovial tissue;Western blot was applied to detect the expression of YAP,phosphorylated YAP(p-YAP),phosphorylated TAZ(p-TAZ),TAZ,and B-cell lymphoma/leukemia 2 associated X protein(Bax)in synovial tissue.Results Compared with the NC group,the Model group showed an increase in Lequesne MG score,serum TNF-αand IL-6 expression,increased expression of YAP and TAZ mRNA,and increased expression of p-YAP/YAP,p-TAZ/TAZ,Bax proteins(P<0.05);compared with the Model group,the low-dose and high-dose OL groups showed a decrease in Lequesne MG score,serum TNF-αand IL-6 expression,YAP and TAZ mRNA expression,and p-YAP/YAP,p-TAZ/TAZ,Bax protein expression(P<0.05);VT103 significantly weakened the protective effect of OL on the synovial tissue of KOA rats(P<0.05).Conclusion OL may alleviate synovial inflammation in knee osteoarthritis rats by activating the YAP/TAZ signaling pathway.