目的 基于热敏通道瞬时受体电位通道香草醛亚型1（TRPV1）/瞬时受体电位销蛋白1（TRPA1）通路探究清脑止痛胶囊治疗偏头疼的作用机制。方法 取Wistar大鼠60只随机分为对照组、模型组、正天丸组（阳性对照，1.8 g·kg^-1）和清脑止痛胶囊低、中、高剂量（0.35、0.70、1.40 g·kg^-1）组。除对照组外，其余各组均用利血平0.7 mg·kg^-1 sc注射法建立偏头痛大鼠模型。各组均于造模成功后ig给予相应剂量药物，每天1次，连续7 d。末次给药结束12 h后，观察各组大鼠挠头次数及行为学变化；采用vonFrey法测试大鼠眶周疼痛阈值；酶联免疫法检测脑组织中白细胞介素-1（IL-1）、肿瘤坏死因子-α（TNF-α）、5-羟色胺（5-HT）水平变化；试剂盒法检测脑组织中一氧化氮（NO）、钙离子（Ca²⁺）水平；免疫荧光法检测TRPV1、TRPA1与神经元微管相关蛋白2（MAP2）在脑组织中共表达水平；蛋白免疫印迹（Western blotting）法检测脑组织中TRPV1/TRPA1通路蛋白、降钙素基因相关肽（CGRP）、环氧化酶2（COX-2）表达水平。结果 与模型组比较，清脑止痛胶囊低、中、高剂量组及正天丸组大鼠挠头次数，脑干组织NO、Ca²⁺、IL-1及TNF-α水平，TRPV1、TRPA1分别与MAP2阳性神经元共表达水平，TRPV1、TRPA1、CGRP、COX-2蛋白表达水平均显著降低（P<0.05），眶周疼痛阈值、5-HT表达显著升高（P<0.05），且上述指标变化均呈剂量相关性，高剂量组与正天丸组相比差异无统计学意义。结论 清脑止痛胶囊可能通过降低TRPV1/TRPA1通路介导的疼痛信号传导，改善偏头痛大鼠疼痛症状。

Objective To explore the mechanism of Qingnao Zhitong Jiaonang (QNZTJN) in the treatment of migraine based on transient receptor potential Vanilloild-1 (TRPV1)/transient receptor potential ankyrin-1 (TRPA1) pathway. Methods A total of 60 Wistar rats were randomly divided into control group, model group, positive control group (Zhengtian Pill, 1.8 g·kg^-1), QNZTJN low, middle, and high dose (0.35, 0.70, 1.40 g·kg^-1) groups. Except the control group, the other groups were injected with reserpine 0.7 mg·kg^-1 subcutaneously to establish migraine rat models. After successful modeling, the rats in each group were given corresponding doses of drugs by gavage for seven days, once a day. At 12 hours after the end of the last administration, the number of head scratching and behavioral changes of rats in each group were observed; the threshold of periorbital pain was measured by vonFrey method; the levels of interleukin-1 (IL-1), tumor necrosis factor-α (TNF-α) and 5-hydroxytryptamine (5-HT) in brain tissue were detected by enzyme linked immunosorbent assay; the levels of nitric oxide (NO) and calcium ion (Ca²⁺) in brain tissue were detected by kit method; the co expression levels of TRPV1, TRPA1 with neuronal microtubule associated protein 2 (MAP2) were detected by immunofluorescence; the expression levels of TRPV1/TRPA1 pathway protein, calcitonin gene related peptide (CGRP) and cyclooxygenase 2 (COX-2) were detected by Western blotting. Results Compared with model group, the number of head scratching, the levels of NO, Ca²⁺, IL-1 and TNF-α, the co-expression levels of TRPV1 and TRPA1 with MAP2 positive neurons, and the protein expression of TRPV1, TRPA1, CGRP and COX-2 decreased in the QNZTJN low, medium, high dose groups and positive control group (P < 0.05), the periorbital pain threshold and expression of 5-HT increased (P < 0.05), and the changes of above indexes in QNZTJN groups were dose-dependent. There was no significant difference between QNZTJN high-dose group and positive control group. Conclusion QNZTJN may improve the pain symptoms of migraine rats by reducing the pain signal transduction mediated by TRPV1/TRPA1 pathway.

R285.5

吉林省科技发展计划项目、清脑止痛胶囊大品种二次开发（20140310001YY）