P2Y14受体参与小胶质细胞介导的偏头痛中枢敏化

P2Y14 receptor is involved in microglia-mediated migraine central sensitization

摘要:

目的： 三叉神经脊束尾核中的P2X4受体通过激活小胶质细胞从而参与慢性偏头痛的发展已经得到研究证实，但对于P2Y14受体在偏头痛中的作用，尤其是与中枢敏化相关的研究却非常少。我们研究开展的目的是在硬脑膜注射“炎性汤”（IS）和P2Y14抑制剂4,7-二取代的二萘酸衍生物（4,7-disubstituted 2naphthoic acid derivative ，PPTN)后的偏头痛大鼠模型里探索其颈髓C1-C2内P2Y14受体和小胶质细胞的表达情况，从形态和功能上分析它们对中枢敏化发生发展的影响。方法：42只雄性SD大鼠随机分为生理盐水（NS）组、“炎性汤”4天（IS4d）组、“炎性汤”7天（IS7d）组、PPTN预防组、预防对照组、PPTN治疗组和治疗对照组。Von Frey纤维丝测定大鼠的眶周痛阈值，Western Blot检测颈髓后角P2Y14和Iba-1蛋白的表达情况，免疫荧光检测颈髓后角Iba-1数量及形态变化【建议3】。结果：与NS组相比，IS4d组和IS7d组大鼠眶周痛阈值明显下降，而PPTN预防组和PPTN治疗组眶周痛阈值的下降程度均远小于各自相对应的NS预防对照组和NS治疗对照组（p<0.01）。Western Blot结果显示，P2Y14受体和小胶质细胞特异性蛋白Iba-1含量在各组的变化趋势与眶周痛阈值呈一致性，免疫荧光结果亦然【建议3】。结论：P2Y14受体可能通过激活小胶质细胞从而参与偏头痛中枢敏化的发生发展，其抑制剂PPTN可以起到预防和治疗的作用，不仅可以降低中枢敏化的程度，减缓中枢敏化的进程，甚至可能抑制中枢敏化的出现。

Abstract:

Objective P2X4 receptor in the trigeminal nucleus caudalis (TNC) activates microglia to participate in the development of chronic migraine has been demonstrated. However, very little research has been done on the role of P2Y14 receptor in migraine, especially in relation to central sensitization. The purpose of our study was to explore the expression of P2Y14 receptor and microglia in C1-C2 of spinal cord after dural injection of inflammatory decoction (IS) and P2Y14 inhibitor PPTN in migraine rats, and to analyze their influence on the occurrence and development of central sensitization in terms of morphology and function. Methods 42 male SD rats were randomly divided into normal saline (NS) group, inflammatory soup 4-day (IS4d) group, inflammatory soup 7-day (IS7d) group, PPTN prevention group, prevention control group, PPTN treatment group and treatment control group. Von Frey filaments were used to determine the periorbital pain threshold of rats, and Western Blot was used to detect the expression of P2Y14 and Iba-1 proteins in the posterior horn of cervical spinal cord. Meanwhile, immunofluorescence was used to detect the quantity and morphology of Iba-1【建议3】. Results Compared with the NS group, the periorbital pain threshold of the IS4d group and IS7d group significantly decreased, while the decrease degree of the periorbital pain threshold of the PPTN prevention group and the PPTN treatment group was much lower than their corresponding prevention control group and treatment control group (p<0.01). Western Blot results showed that the variation trend of P2Y14 receptor and microglia specific protein Iba-1 in each group was consistent with the periorbital pain threshold. So did immunofluorescence results【建议3】. Conclusion P2Y14 receptor may be involved in the occurrence and development of migraine central sensitization by activating microglia, and its inhibitor PPTN could play a preventive and therapeutic role, not only reducing the degree of central sensitization, slowing down the process of central sensitization, but even inhibiting the occurrence of central sensitization.