CP-25 combined with MTX/ LEF ameliorates the progression of adjuvant-induced arthritis by the inhibition on GRK2 translocation.
CP-25 combined with MTX/ LEF ameliorates the progression of adjuvant-induced arthritis by the inhibition on GRK2 translocation.
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DOI：
10.1016/j.biopha.2018.12.040
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发表时间：
2019-02
期刊：
影响因子：
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通讯作者：
Xuezhi Yang;Yingjie Zhao;X. Jia;Chun Wang;Yujing Wu;Lingling Zhang;Yan Chang;Wei Wei-Wei
中科院分区：
文献类型：
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作者：
Xuezhi Yang;Yingjie Zhao;X. Jia;Chun Wang;Yujing Wu;Lingling Zhang;Yan Chang;Wei Wei-Wei
在动物模型中，GSCP-25通过抑制G蛋白偶联受体激酶2（GRK 2）膜表达来减弱关节炎进展。本研究比较了治疗组与高剂量甲氨蝶呤（MTX）/来氟米特（LEF）和CP-25结合低剂量MTX/LEF在佐剂诱导的关节炎（AA）大鼠模型和调查可能的mechanism.MethodsAA诱导大鼠通过完全弗氏佐剂。实验组分为正常组;媒介物组;单一疗法组，包括CP-25（50 mg/kg）、MTX（0.25、0.5 mg/kg）和LEF（5、10 mg/kg）;和CP-25（50 mg/kg）-组合MTX（0.25 mg/kg）/LEF（5 mg/kg）组。我们测量了细胞因子水平，磷酸化和蛋白质表达，以及蛋白质之间的相互作用。GRK 2对磷酸化细胞外信号调节激酶（p-ERK）的作用，通过GRK 2-siRNA使用高含量imaging system.ResultsTherapeutic效果，包括病理学和细胞因子平衡，确定CP-25组合组和高剂量MTX/LEF组之间是等效的。在治疗组中，AA成纤维细胞样滑膜细胞（FLS）中的P38、ERK和c-Jun N-末端激酶（JNK）活化减少; GRK 2表达仅在CP-25组中受到抑制。GRK 2和p-ERK之间的相互作用在媒介物组中减少，并且在CP-25组中恢复。GRK 2膜表达和p-ERK核表达在用肿瘤坏死因子α预处理并用前列腺素E2刺激的FLS中增加。结论CP-25联合MTX/LEF组与高剂量MTX/LEF组疗效相当。CP-25通过降低FLS中GRK 2的膜表达来抑制p-ERK。
ObjectiveCP-25 attenuates arthritis progression in animal models by inhibiting G protein-coupled receptor kinase 2 (GRK2) membrane expression. This study compared groups treated with high-dose methotrexate (MTX)/leflunomide (LEF) and CP-25 combined with low-dose MTX/LEF in an adjuvant-induced arthritis (AA) rat model and investigated possible mechanisms.MethodsAA was induced in rats via complete Freund’s adjuvant. Experimental groups were divided into a normal group; vehicle group; monotherapy groups, including CP-25 (50 mg/kg), MTX (0.25, 0.5 mg/kg), and LEF (5, 10 mg/kg); and CP-25 (50 mg/kg)-combined MTX (0.25 mg/kg)/LEF (5 mg/kg) groups. We measured cytokine levels, phosphorylation and protein expression, and interactions between proteins. The role of GRK2 on phosphorylated extracellular signal-regulated kinase (p-ERK) was determined via GRK2-siRNA using a high content imaging system.ResultsTherapeutic effects, including pathology and cytokine balance, were equivalent between the CP-25-combination groups and the high-dose MTX/LEF groups. P38, ERK, and c-Jun N-terminal kinase (JNK) activation in AA fibroblast-like synoviocytes (FLS) was reduced in the treatment groups; GRK2 expression was only inhibited in the CP-25 group. Interactions between GRK2 and p-ERK decreased in the vehicle group and were restored in the CP-25 group. GRK2 membrane expression and p-ERK nuclear expression increased in FLS pre-treated with tumour necrosis factor alpha and stimulated with prostaglandin E2. Nuclear expression of p-ERK increased in GRK2-siRNA FLS.ConclusionEquivalent therapeutic effects were observed between CP-25-combination groups and high-dose MTX/LEF groups. CP-25 inhibited p-ERK by reducing the membrane expression of GRK2 in FLS from AA rats.