目的 基于熱敏通道瞬時(shí)受體電位通道香草醛亞型1（TRPV1）/瞬時(shí)受體電位銷蛋白1（TRPA1）通路探究清腦止痛膠囊治療偏頭疼的作用機(jī)制。方法 取Wistar大鼠60只隨機(jī)分為對照組、模型組、正天丸組（陽性對照，1.8 g·kg^-1）和清腦止痛膠囊低、中、高劑量（0.35、0.70、1.40 g·kg^-1）組。除對照組外，其余各組均用利血平0.7 mg·kg^-1 sc注射法建立偏頭痛大鼠模型。各組均于造模成功后ig給予相應(yīng)劑量藥物，每天1次，連續(xù)7 d。末次給藥結(jié)束12 h后，觀察各組大鼠撓頭次數(shù)及行為學(xué)變化；采用vonFrey法測試大鼠眶周疼痛閾值；酶聯(lián)免疫法檢測腦組織中白細(xì)胞介素-1（IL-1）、腫瘤壞死因子-α（TNF-α）、5-羥色胺（5-HT）水平變化；試劑盒法檢測腦組織中一氧化氮（NO）、鈣離子（Ca²⁺）水平；免疫熒光法檢測TRPV1、TRPA1與神經(jīng)元微管相關(guān)蛋白2（MAP2）在腦組織中共表達(dá)水平；蛋白免疫印跡（Western blotting）法檢測腦組織中TRPV1/TRPA1通路蛋白、降鈣素基因相關(guān)肽（CGRP）、環(huán)氧化酶2（COX-2）表達(dá)水平。結(jié)果 與模型組比較，清腦止痛膠囊低、中、高劑量組及正天丸組大鼠撓頭次數(shù)，腦干組織NO、Ca²⁺、IL-1及TNF-α水平，TRPV1、TRPA1分別與MAP2陽性神經(jīng)元共表達(dá)水平，TRPV1、TRPA1、CGRP、COX-2蛋白表達(dá)水平均顯著降低（P<0.05），眶周疼痛閾值、5-HT表達(dá)顯著升高（P<0.05），且上述指標(biāo)變化均呈劑量相關(guān)性，高劑量組與正天丸組相比差異無統(tǒng)計(jì)學(xué)意義。結(jié)論 清腦止痛膠囊可能通過降低TRPV1/TRPA1通路介導(dǎo)的疼痛信號傳導(dǎo)，改善偏頭痛大鼠疼痛癥狀。

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

吉林省科技發(fā)展計(jì)劃項(xiàng)目、清腦止痛膠囊大品種二次開發(fā)（20140310001YY）