基于JAK/STAT信号通路探讨复方竹节参片对膝关节骨性关节炎兔的作用机制

目的探讨JAK/STAT信号通路在膝关节骨性关节炎(KOA)发生、发展和转归中的作用,初步揭示复方竹节参片治疗KOA的作用环节和靶点。方法取6只雄性日本大耳白兔作为对照组,取34只雄性日本大耳白兔采用关节腔注射木瓜蛋白酶诱导KOA模型。将造模成功的兔随机分为模型组8只和复方竹节参片高、中、低剂量组各6只。造模结束后,复方竹节参片高、中、低剂量组分别按150 mg/kg、75 mg/kg、37.5 mg/kg的剂量灌胃给予复方竹节参片混悬液,模型组和对照组同法给予蒸馏水,均每天1次,连续14 d。分别于造模前、造模结束后、灌胃7 d和灌胃14 d测量各组兔脚踝周径;于末次灌胃后24 h,取膝关节、血清和滑膜组织,采用HE染色法观察膝关节组织病理学形态,ELISA法测定血清中白细胞介素-6(IL-6)水平,Western blot法测定滑膜组织中JAK2、p-JAK、STAT3、ERK1蛋白表达量。结果与对照组比较,模型组兔脚踝周径显著增加(P<0.05),血清IL-6水平及滑膜中JAK2、p-JAK、STAT3、ERK1蛋白表达量均显著升高(P均<0.05)。与模型组比较,复方竹节参片各组兔脚踝周径明显缩短(P均<0.05),复方竹节参片高、中剂量组血清IL-6水平均显著降低(P均<0.05);复方竹节参片高剂量组滑膜组织中JAK2蛋白表达量及复方竹节参片各组滑膜组织中p-JAK2、STAT3、ERK1蛋白表达量均显著降低(P均<0.05)。结论复方竹节参片能显著缓解KOA的关节肿胀,其机制可能与调控JAK/STAT信号通路上下游相关的细胞因子和蛋白表达有关。

Discussion on the mechanism of compound panax japonicus tablets in the treatment of rabbits with knee osteoarthritis based on JAK/STAT signal pathway

Objective It is to explore the role of JAK/STAT signaling pathway in the occurrence,development and outcome of knee osteoarthritis(KOA),and preliminarily reveal the role and target point of compound panax japonicus tablets in the treatment of KOA.Methods Six male Japanese white rabbits were taken as the control group,and another 34 male Japanese white rabbits were injected with papain into the joint cavity to induce the KOA models.The successfully modeled rabbits were randomly divided into a model group with 8 rabbits and compound panax japonicus tablet high,medium,and low dose groups with 6 rabbits in each group.At the end of modeling,the high,medium,and low dose groups of compound panax japonicus tablets were given a suspension of compound panax japonicus tablets at the doses of 150 mg/kg,75 mg/kg,and 37.5 mg/kg by gavage,respectively.The model group and the control group were given distilled water by gavage,all once a day for 14 consecutive days.The ankle circumferences of the rabbits in each group were measured before and after the modeling,7 days and 14 days after gavage;at 24 hours after the last gavage,the knee joints,serum and synovial tissue were taken to observe the histopathological morphology of knee joint by HE staining,to determine the levels of serum interleukin-6(IL-6)by ELISA method,and to detect the expression of JAK2,p-JAK,STAT3,ERK1 protein in synovial tissue by Western blot method.Results Compared with the control group,the length of ankle circumference,the content of serum IL-6 and the expressions of STAT3,JAK2,p-JAK2 and ERK1 proteins in synovium in the model group were significantly increased(all P<0.05).Compared with the model group,the length of ankle circumference in the high,medium and low dose groups of compound panax japonicus tablets was significantly shortened(all P<0.05),the content of serum IL-6 in high and medium dose groups of compound panax japonicus tablets was significantly decreased(P<0.05),JAK2 protein expression in synovium of compound panax japonicus high dose group and the expression of ErK1,STAT3,p-JAK2 protein in synovium of each treatment group of compound panax japonicus tablets was significantly decreased(all P<0.05).Conclusion Compound panax japonicus tablets can significantly relieve the joint swelling of KOA,its mechanism may be related to the regulation of cytokine and protein expression upstream and downstream related to the JAK/STAT signaling pathway.