由IL-10/JAK2/STAT3通路探讨养血柔筋方对兔膝骨关节炎软骨损伤的研究

Exploring the effect of Yangxue Roujin formula on cartilage injury in rabbit knee osteoarthritis through the IL-10/JAK2/STAT3 pathway

目的 由白细胞介素(IL)-10/Janus激酶(JAK2)/信号传导与转录激活因子3(STAT3)通路探讨养血柔筋方对兔膝骨关节炎(knee osteoarthritis, KOA)软骨损伤的影响及机制。方法 将新西兰雄兔分为空白组、KOA模型组、养血柔筋方组(50 mg/kg)、塞来昔布组(24 mg/kg)、JAK2激活剂组(50 mg/kg养血柔筋方+0.11 mg/kg AG490),每组6只。通过木瓜蛋白酶诱导KOA兔模型,造模成功后给予相应治疗。运用HE染色观察软骨组织病理变化,Mankin评分进行定量评分;透射电镜观察软骨组织的超微结构;ELISA测量血清IL-1β、IL-6含量;免疫组织化学检测软骨组织基质金属蛋白酶13(MMP13)、转化生长因子-β_(1)(TGF-β_(1))、Ⅱ型胶原蛋白(COl-Ⅱ)蛋白表达;Western Blot检测软骨组织IL-10/JAK2/STAT3通路蛋白表达。结果 与空白组比较,KOA模型组中软骨组织病变严重,Mankin评分、IL-1β和IL-6含量、MMP13、p-JAK2和p-STAT3蛋白水平上调,TGF-β_(1)、COl-Ⅱ和IL-10蛋白水平均下调(P<0.05)。与KOA模型组比较,养血柔筋方组、塞来昔布组关节软骨病理损伤明显恢复,Mankin评分、IL-1β和IL-6含量、MMP13、p-JAK2和p-STAT3蛋白水平下调,TGF-β_(1)、COl-Ⅱ和IL-10蛋白水平均上调(P<0.05)。JAK2激活剂可明显削弱养血柔筋方对KOA模型的影响(P<0.05)。结论 养血柔筋方可能通过调节IL-10/JAK2/STAT3通路从而抑制KOA兔软骨损伤。

Objective Based on the interleukin(IL)-10/Janus kinase(JAK2)/signal transduction and activator of transcription 3(STAT3)pathway,to explore the effect and mechanism of Yangxue Roujin formula on cartilage damage in rabbits with knee osteoarthritis(KOA).Methods New Zealand male rabbits were divided into blank group,KOA model group,Yangxue Roujin formula group(50 mg/kg),celecoxib group(24 mg/kg),and JAK2 activator group(50 mg/kg Yangxue Roujin formula+0.11 mg/kg AG490),6 per group.The KOA rabbit model was induced by papain,and the corresponding treatment was given after the model was successfully established.HE staining was applied to observe the pathological changes of cartilage tissue,Mankin score was used for quantitative scoring;transmission electron microscopy was applied to observe the ultrastructure of cartilage tissue;ELISA was applied to measure the levels of serum IL-1βand IL-6;immunohistochemistry was applied to detect the expression of cartilage tissue matrix metalloproteinase 13(MMP13),transforming growth factor-β_(1)(TGF-β_(1)),collagen type II(COl-II)proteins;and Western blot was applied to detect the expression of IL-10/JAK2/STAT3 pathway proteins in cartilage tissue.Results Compared with the blank group,the cartilage tissue lesions in the KOA model group were severe,the Mankin score,the contents of IL-1βand IL-6,and the protein levels of MMP13,p-JAK2 and p-STAT3 were up-regulated,the protein levels of TGF-β_(1),COl-II and IL-10 were all down-regulated(P<0.05).Compared with the KOA model group,the pathological damage of the articular cartilage in the Yangxue Roujin recipe group and the celecoxib group recovered obviously,the Mankin score,the contents of IL-1βand IL-6,and the protein levels of MMP13,p-JAK2 and p-STAT3 were down-regulated,the protein levels of TGF-β_(1),COl-II and IL-10 were all up-regulated(P<0.05).JAK2 activator was able to obviously weaken the effect of Yangxue Roujin formula on KOA model(P<0.05).Conclusion Yangxue Roujin formula may inhibit cartilage injury in KOA rabbits by regulating IL-10/JAK2/STAT3 pathway.