芍药苷-6′-O苯磺酸酯调节GRK2对CIA大鼠巨噬细胞JAK1/STAT3信号通路的影响

Effect of paeoniflorin-6′-O benzene sulfonate on JAK1/STAT3 signaling pathway in rat macrophages with collagen-induced arthritis through regulating G protein coupled receptor kinase 2

目的明确芍药苷-6′-O苯磺酸酯(代号CP-25)调节G蛋白偶联受体激酶2(G protein coupled receptor kinase 2,GRK2)发挥对胶原性关节炎(collagen-induced arthritis,CIA)大鼠巨噬细胞JAK1/STAT3信号通路的抑制作用。方法SD雄性大鼠随机分为4组:正常组、模型组、CP-25(50 mg·kg-1·d-1)组、MTX(0.5 mg·kg-1·3d-1)组。在造模后连续21 d灌胃给药。观察检测大鼠整体指标,检测大鼠腹腔巨噬细胞培养上清和外周血血清中IL-6、PGE2的水平、CD86/CD206的比值、GRK2的胞膜表达及GRK2与JAK1共定位、JAK1/STAT3蛋白表达及p-STAT3入核情况。提取正常大鼠腹腔巨噬细胞,体外IL-6、PGE2刺激24 h后,分别检测上述指标。结果CP-25可明显改善大鼠关节肿胀和全身炎症情况,降低大鼠腹腔巨噬细胞培养上清和外周血中IL-6和PGE2的水平,CP-25明显降低巨噬细胞CD86/CD206、JAK1/STAT3蛋白表达及GRK2胞膜表达、p-STAT3入核,增加GRK2与JAK1的共定位结合率。结论CP-25对CIA大鼠发挥治疗作用,其重要作用机制之一是通过调节GRK2活性,进而抑制JAK1/STAT3信号通路。

Aim To investigate the inhibitory effect of peonia-6′-o benzene sulfonate(CP-25)on the JAK1/STAT3 signaling pathway in collagen-induced arthritis(CIA)rats macrophages through regulating G protein coupled receptor kinase 2(GRK2).Methods SD rats were randomly divided into four groups:normal group,model group,CP-25(50 mg·kg-1·d-1)group and MTX(0.5 mg·kg-1·3d-1)group.The drug was administered orally for 21 consecutive days after modeling.During the administration,the overall index of the rats were observed,the levels of IL-6 and PGE2 in peritoneal macrophage culture supernatant and peripheral blood serum were detected,and the ratio of CD86/CD206,the cell membrane expression of GRK2 and the co-localition of GRK2 and JAK1,the protein expression of JAK1/STAT3 and the nuclear translocation of p-STAT3 were all detected.The normal rat peritoneal macrophages were extracted,and after IL-6 and PGE2 were stimulated for 24 h in vitro,the above indexes were detected respectively.Results CP-25 could significantly relieve joint swelling and systemic inflammation in CIA rats,CP-25 could significantly reduce the levels of IL-6 and PGE2 in culture supernatant and peripheral blood serum of rat peritoneal macrophages.CP-25 could significantly reduce the expression of CD86/CD206,JAK1/STAT3 protein,GRK2 membrane expression and p-STAT3 nuclear translocation of macrophages,and increase the co-localization rate of GRK2 and JAK1.Conclusions CP-25 has a therapeutic effect on CIA rats,and one of its important mechanisms is inhibiting the JAK1/STAT3 signaling pathway by regulating GRK2 activity.