依达拉奉调节HIF-1α/VEGF信号通路对类风湿性关节炎大鼠血管生成及炎症反应的影响

Impacts of edaravone on angiogenesis and inflammatory reaction in rheumatoid arthritis rats by regulating the HIF-1α/VEGF signaling pathway

目的探讨依达拉奉(Eda)调节缺氧诱导因子-1α(HIF-1α)/血管内皮生长因子(VEGF)信号通路对类风湿性关节炎(RA)大鼠血管生成及炎症反应的影响。方法将60只6~8周龄SPF级雄性SD大鼠分为control组、RA组、Eda组、甲氨蝶呤组、二甲基草酰甘氨酸(DMOG)组(HIF-1α/VEGF通路激活剂),每组12只,除control组外均采用Ⅱ型胶原蛋白联合完全弗氏佐剂复制RA大鼠模型。所有干预结束后,检测大鼠足趾肿胀度、痛阈值并对大鼠进行关节炎评分;ELISA法检测血清白细胞介素-6(IL-6)、肿瘤坏死因子(TNF-α)水平;CD31免疫组化检测足关节滑膜组织血管密度;HE染色观察足关节滑膜组织病理改变;qRT-qPCR法检测HIF-1α、VEGF mRNA表达;Western blot法检测HIF-1α/VEGF信号通路蛋白表达。结果相较于control组,RA组大鼠足肿胀度、关节炎评分、IL-6、TNF-α水平、足关节滑膜组织血管密度、HIF-1α、VEGF mRNA及蛋白表达升高,压痛阈值、热痛阈值降低(P<0.05);相较于RA组,Eda组与甲氨蝶呤组大鼠足肿胀度、关节炎评分、IL-6、TNF-α水平、足关节滑膜组织血管密度、HIF-1α、VEGF mRNA及蛋白表达降低,压痛阈值、热痛阈值升高(P<0.05);HIF-1α/VEGF通路激活剂DMOG可减弱Eda对RA大鼠血管生成及炎症反应的抑制作用(P<0.05)。结论Eda通过抑制HIF-1α/VEGF信号通路,抑制血管生成、降低炎症反应,从而起到减轻RA的作用。

Objective To investigate the impacts of edaravone(Eda)on angiogenesis and inflammatory reaction in rheumatoid arthritis(RA)rats by regulating the hypoxia inducible factor-1α(HIF-1α)/vascular endothelial growth factor(VEGF)signaling pathway.Methods 60 rats were separated into control group,RA group,Eda group,methotrexate group,and dimethyloxaloglycine(DMOG)group(HIF-1α/VEGF pathway activator),with 12 rats in each group.Except for the control group,RA rat model was replicated using typeⅡcollagen combined with complete Freund's adjuvant.After all interventions were completed,the degree of foot swelling and pain threshold of rats were measured,and arthritis scores were performed on the rats.ELISA method was applied to detect serum levels of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α).CD31 immunohistochemistry was applied to detect the vascular density of synovial tissue in the foot joint.HE staining was applied to observe pathological changes in the synovial tissue of the foot joint.QRT-qPCR method was applied to detect HIF-1αand VEGF mRNA expression.Western blot was applied to detect the expression of HIF-1α/VEGF signaling pathway proteins.Results Compared to the control group,the foot swelling degree,arthritis score,IL-6,TNF-αlevels,vascular density of synovial tissue in the foot joints,HIF-1α,VEGF mRNA and protein expression increased in RA group,while the tenderness threshold and heat pain threshold decreased(P<0.05).Compared to the RA group,the foot swelling degree,arthritis score,IL-6,TNF-αlevels,vascular density of synovial tissue in the foot joints,HIF-1α,VEGF mRNA and protein expression decreased in Eda group and methotrexate group,while the tenderness threshold and heat pain threshold increased(P<0.05).The HIF-1α/VEGF pathway activator DMOG was able to weaken the inhibitory effect of Eda on angiogenesis and inflammatory reaction in RA rats(P<0.05).Conclusion Eda can inhibit angiogenesis and reduce inflammatory reaction by inhibiting the HIF-1α/VEGF signaling pathway,thereby reducing RA.