Abstract：
Nowadays, the pharmacological management of visceral hypersensitivity associated with colitis is ineffective. In this context, targeting purinergic P2X4 receptor (P2X4R), which can modulate visceral pain transmission, could represent a promising therapeutic strategy. Herein, we tested the pain-relieving effect of two novel and selective P2X4R antagonists (NC-2600 and NP-1815-PX) in a murine model of DNBS-induced colitis and investigated the mechanisms underlying their effect. Tested drugs and dexamethasone (DEX) were administered orally, two days after colitis induction. Treatment with tested drugs and DEX improved tissue inflammatory parameters (body weight, spleen weight, macroscopic damage, TNF and IL-1β levels) in DNBS-rats. In addition, NC-2600 and NP-1815-PX attenuated visceral pain better than DEX and prevented the reduction of occludin expression. In in vitro studies, treatment of CaCo2 cells with supernatant from THP-1 cells, previously treated with LPS plus ATP, reduced the expression of tight junctions protein. By contrast, CaCo2 cells treated with supernatant from THP-1 cells, previously incubated with tested drugs, counteracted the reduction of tight junctions due to the inhibition of P2X4R/NLRP3/IL-1β axis. In conclusion, these results suggest that the direct and selective inhibition of P2X4R represents a viable approach for the management of visceral pain associated with colitis via NLRP3/IL-1β axis inhibition.
摘要：
如今,与结肠炎相关的内脏过敏的药物治疗无效.在这种情况下，靶向嘌呤能P2X4受体（P2X4R），可以调节内脏疼痛的传播，可以代表一种有前途的治疗策略。在这里，我们在DNBS诱导的结肠炎小鼠模型中测试了两种新型选择性P2X4R拮抗剂(NC-2600和NP-1815-PX)的疼痛缓解作用,并研究了其作用机制.试验药物和地塞米松（DEX）口服给药，结肠炎诱导后两天。用测试药物和DEX治疗可改善组织炎症参数（体重，脾脏重量,宏观损伤，DNBS大鼠的TNF和IL-1β水平）。此外,NC-2600和NP-1815-PX比DEX更好地减轻内脏疼痛，并防止了occludin表达的减少。在体外研究中，用THP-1细胞的上清液处理CaCo2细胞，以前用LPS加ATP治疗，降低紧密连接蛋白的表达。相比之下,用THP-1细胞的上清液处理的CaCo2细胞，之前与测试药物一起孵育，由于P2X4R/NLRP3/IL-1β轴的抑制而抵消了紧密连接的减少。总之,这些结果表明,直接和选择性抑制P2X4R是通过抑制NLRP3/IL-1β轴治疗结肠炎相关内脏痛的可行方法.
