摘要
目的:探讨Janus激酶2/信号转导及转录激活因子3(JAK2/STAT3)信号通路在膝骨关节炎(KOA)大鼠滑膜组织中的表达及其对血管内皮生长因子(VEGF)和干细胞生长因子-β(SCGF-β)的调控作用。方法:将40只SD雌性大鼠随机分为4组,即正常对照组、模型组、AG490组和DUSP19组,每组10只。除正常对照组外,其他3组均通过左膝关节腔内注射L-半胱氨酸—木瓜蛋白酶混合液建立KOA大鼠模型。造模12 d后,AG490组和DUSP19组分别在关节腔注射JAK2/STAT3信号通路抑制剂AG490和激活剂DUSP19,正常对照组和模型组注射等量生理盐水。取各组滑膜组织,苏木精—伊红(HE)染色行滑膜组织病理学检查,采用RT-qPCR法检测JAK2、STAT3、SCGF-β、VEGF mRNA表达;Western blotting法检测p-JAK2、p-STAT3、SCGF-β、VEGF蛋白表达。采用Pearson相关分析方法分析p-JAK2、p-STAT3与SCGF-β、VEGF表达的相关性。结果:与正常对照组比较,模型组滑膜细胞表面明显肿胀,存在炎细胞浸润,增生活跃,毛细血管增多且充血明显;与模型组比较,AG490组滑膜细胞异常增生、排列情况、炎细胞浸润均显著恶化,而DUSP19组细胞形态显著改善。与正常对照组比较,模型组JAK2、STAT3mRNA及p-JAK2、p-STAT3蛋白表达量显著降低,SCGF-β、VEGF mRNA及蛋白表达量显著升高(均P<0.05);与正常对照组和模型组比较,AG490组JAK2、STAT3 m RNA及p-JAK2、p-STAT3蛋白表达量显著降低,SCGF-β、VEGF mRNA及蛋白表达量显著升高(均P<0.05);相反地,DUSP19组JAK2、STAT3 mRNA及p-JAK2、p-STAT3蛋白表达量显著升高,SCGF-β、VEGF mRNA及蛋白表达量显著降低(均P<0.05)。Pearson相关分析显示,p-JAK2、p-STAT3表达与SCGF-β、VEGF表达呈负相关关系(均P<0.05)。结论:JAK2/STAT3信号通路在KOA大鼠滑膜组织中的表达被显著抑制,该通路的激活可下调SCGF-β、VEGF的表达而减轻KOA大鼠关节病理损伤。
Objective: To investigate the expression of Janus kinase2/signal transduction and activator of transcription 3(JAK2/STAT3) signaling pathway in the synovial tissues of knee osteoarthritis(KOA)rats and its regulation on vascular endothelial growth factor(VEGF) and stem cell growth factor-β(SCGF-β) expression. Methods: Forty SD female rats were randomly divided into normal control group, model group, AG490 group and DUSP19 group, with 10 rats in each group. Except for the normal control group, the rats in other three groups were injected with L-cysteine-papa in mixture via left knee joint to establish KOA models. After 12 days of modeling, AG490 and DUSP19 were injected into the articular cavity of rats in AG490 group and DUSP19 group, respectively. Rats in the normal control group and model group were injected with same volume of normal saline.The m RNA expressions of JAK2, STAT3, SCGF-β and VEGF were detected by RT-qPCR. The protein expressions of p-JAK2, p-STAT3, SCGF and VEGF were determined by Western blotting. The correlation between the expressions of p-JAK2, p-STAT3, SCGF-β and VEGF was evaluated by Pearson correlation analysis. Results: Compared with the normal control group, evidently swollen synovial cell surface, massive inflammatory cell infiltration, synovial hyperplasia, increased capillary and obvious hyperemia were observed in the model group. Compared with the model group, the synovial hyperplasia, disordered cell arrangement, and inflammatory cell infiltration were significantly worsened in AG490 group, while the morphology of cells was significantly improved in DUSP19 group. The JAK2 and STAT3 mRNA levels and p-JAK2 and p-STAT3 protein levels in the model group were lower than those in the normal control group, while the mRNA and protein levels of SCGF-β and VEGF were markedly higher(P<0.05).Compared with normal control group and model group, the expressions of JAK2 and STAT3 mRNA and p-JAK2 andp-STAT3 protein in AG490 group were significantly decreased, while the expressions of SCGF-β and VEGF mRNA and protein were increased(P<0.05). However, opposite effects were noted in DUSP group(P<0.05).Pearson correlation analysis showed that p-JAK2 and p-STAT3 expressions were negatively correlated with SCGF-β and VEGF expressions(P<0.05). Conclusion: The JAK2/STAT3 signaling pathway was significantly inhibited in synovial tissues of KOA rats;activation of JAK2/STAT3 pathway down-regulated the expression of SCGF-β and VEGF, and alleviated joint pathological injury in KOA rats.
作者
董朝军
刘宣毅
李冕
东彬
杨晓雅
Dong Chaojun;Liu Xuanyi;Li Mian;Dong Bin;Yang Xiaoya(Cangzhou People's Hospital of Hebei Province,Cangzhou 061000,China)
出处
《广西医科大学学报》
CAS
2021年第9期1735-1740,共6页
Journal of Guangxi Medical University
基金
河北省重点研发计划项目(No.V1585054099367)。
作者简介
通信作者:杨晓雅。
