Ahmad Sheikh Fayaz, Ansari Mushtaq Ahmad, Nadeem Ahmed, Zoheir Khairy M A, Bakheet Saleh A, Al-Shabanah Othman A, Al Rikabi Ammar Cherkess, Attia Sabry M
Department of Pharmacology and Toxicology, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia.
Department of Pharmacology and Toxicology, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia.
Mol Immunol. 2016 Oct;78:65-78. doi: 10.1016/j.molimm.2016.08.017. Epub 2016 Sep 5.
Protein tyrosine kinases are key mediators of the signal transduction cascades that control expression of many genes involved in the induction of inflammation caused by arthritis. Here we investigate the effect of the tyrosine kinase inhibitor tyrphostin AG126 on a mouse model of adjuvant-induced arthritis (AIA). We report that when given at 5mg/kg i.p. every 48h from days 0-21, AG126 exerts potent anti-arthritic effects. Further, we investigated the role of AG126 on the key mediators of arthritic inflammation, namely, edema, arthritic score, presence of immunophenotypes including Foxp3, CD4Foxp3, and CD25Foxp3 T regulatory (Treg) cells, as well as pro- and anti-inflammatory mediators. AG126 treatment significantly attenuated the severity of AIA and caused a substantial reduction in the percentage of CD2, CD3, CD4, CD8, CD23, CD80, CD86 CD122, CD195, TCRβ, and GITR cells in whole blood. Moreover, administration of AG126 under arthritis-inducing conditions resulted in suppression of IL-17A, IFN-γ, CD4 and CD25 populations while causing an increase in the Foxp3, CD4Foxp3, and CD25Foxp3 Treg populations in the spleen. In addition, RT-PCR analysis revealed increased expression of CD4, CD8, IL-17A, IFN-γ, TNF-α, and NF-κB p65 mRNAs and decreased IL-4 mRNA in the arthritic control (AC) mice, while treatment of animals with AG126 reversed these effects. Western blot analysis confirmed the decreased expression of IL-17, GITR, NF-κB p65 proteins and increased Foxp3 and IL-4 proteins following AG126 treatment of knee tissue. Thus, our findings provide new evidence that inhibition of protein tyrosine kinase activity decreases the progression of arthritis.
蛋白酪氨酸激酶是信号转导级联反应的关键介质，这些级联反应控制着许多与关节炎所致炎症诱导相关的基因的表达。在此，我们研究酪氨酸激酶抑制剂 tyrphostin AG126 对佐剂诱导性关节炎（AIA）小鼠模型的影响。我们报告称，从第 0 天至第 21 天，每 48 小时腹腔注射 5mg/kg 的 AG126 可发挥强大的抗关节炎作用。此外，我们研究了 AG126 对关节炎炎症关键介质的作用，即水肿、关节炎评分、包括 Foxp3、CD4Foxp3 和 CD25Foxp3 T 调节（Treg）细胞在内的免疫表型的存在，以及促炎和抗炎介质。AG126 治疗显著减轻了 AIA 的严重程度，并使全血中 CD2、CD3、CD4、CD8、CD23、CD80、CD86、CD122、CD195、TCRβ 和 GITR 细胞的百分比大幅降低。此外，在诱导关节炎的条件下给予 AG126 可导致脾脏中 IL-17A、IFN-γ、CD4 和 CD25 群体受到抑制，同时使 Foxp3、CD4Foxp3 和 CD25Foxp3 Treg 群体增加。此外，逆转录-聚合酶链反应（RT-PCR）分析显示，关节炎对照（AC）小鼠中 CD4、CD8、IL-17A、IFN-γ、TNF-α 和 NF-κB p65 mRNA 的表达增加，而 IL-4 mRNA 减少，而用 AG126 治疗动物可逆转这些作用。蛋白质印迹分析证实，AG126 治疗膝关节组织后，IL-17、GITR、NF-κB p65 蛋白的表达降低，而 Foxp3 和 IL-4 蛋白增加。因此，我们的研究结果提供了新的证据，表明抑制蛋白酪氨酸激酶活性可降低关节炎的进展。
