芎芷石膏汤对硝酸甘油致偏头痛大鼠血浆NO, NOS, CGRP及脑中5-HT, 5-HIAA含量的影响

目的:研究芎芷石膏汤的镇痛作用及对硝酸甘油致偏头痛大鼠血浆一氧化氮(NO)、一氧化氮合酶(NOS)和降钙素基因相关肽(CGRP)及脑组织中5-羟色胺(5-HT)、5-羟吲哚乙酸(5-HIAA)含量的影响。方法:将小鼠和大鼠分为空白、模型、实验药物3个剂量和阳性对照组(阿司匹林0.2 g·kg-1),以芎芷石膏汤低、中、高剂量(小鼠:6,12,24 g·kg-1·d-1;大鼠:3,6,12 g·kg-1·d-1)灌胃给药干预7 d。福尔马林实验末次给药后30 min后小鼠右后足跖皮下注射浓度为2.5%的福尔马林20μL(正常对照组除外)记录小鼠给福尔马林后第Ⅰ时相(0~5 min)和第Ⅱ时相(10~60 min)内小鼠舔右后足的累计时间。热板法于末次给药后15,30,60,90,120 min,在热盘痛觉测试仪(55±0.5)℃上记录每只小鼠首次舔后足或跳跃时间(s)。硝酸甘油所致大鼠偏头痛实验,末次给药后30 min,除空白对照组大鼠以相应剂量95%乙醇sc外,其余各受试组均sc硝酸甘油注射剂10 mg·kg-1,复制偏头痛模型。分光光度法和放免法测大鼠血浆NO,NOS和CGRP含量,高效液相-电化学法测脑组织中5-HT,5-HIAA含量。结果:与正常组比较,福尔马林实验模型组小鼠右后足跖皮下注射福尔马林后,第Ⅰ时相舔足时间,第Ⅱ时相舔足时间均明显增多。在注射硝酸甘油造模后,与空白对照组相比,模型组30 min内大鼠前肢挠头次数较空白对照组均明显增加(P<0.01),大鼠血浆中的NO,NOS,CGRP明显升高(P<0.01),5-HT明显下降(P<0.05)。芎芷石膏汤能有效抑制福尔马林致小鼠疼痛反应,明显减少小鼠I时相舔足时间和Ⅱ时相舔足时间,与模型组比较差异有统计学意义(P<0.05,P<0.01),并对热板致痛小鼠的物理疼痛镇痛效果明显,能显著提高小鼠的痛阈值,与空白对照组比较差异显著(P<0.05,P<0.01)。硝酸甘油致偏头痛模型组大鼠血浆NO,NOS,CGRP与空白对照组相比均明显增高(P<0.05),而脑中5-HT,5-HIAA含量与空白对照组相比明显下降(P<0.05)。与模型组相比,经不同剂量芎芷石膏汤灌胃后大鼠血浆NO,NOS,CGRP增加受到抑制,同时脑中5-HT向5-HIAA的代谢减少,5-HT的量增加,尤以中、高剂量组明显(P<0.05,P<0.01)。结论:芎芷石膏汤具有明显镇痛作用,可能通过降低血浆中NO,NOS,CGRP含量和升高脑中5-HT含量,发挥对偏头痛的治疗作用。

Effect of Xiongzhi Shigao Tang on Plasma NO, NOS, CGRP and 5-HT, 5-HIAA of Brain in Migraine Model Rats

Objective: To observe the analgesic effect of Xiongzhi Shigao tang and the influence on the level of nitric oxide(NO),nitric oxide synthase(NOS), calcitonin gene-related peptide(CGRP)in blood plasma and 5-hydroxytryptamine(5-HT),5-hydroxyindole acetic acid(5-HIAA)in the brain tissue in the rat models of migraine. Method: The mice and rats were divided into blank control group,model group,three doses groups of Xiongzhi Shigao tang (mice:6,12,24 g·kg^-1; rat:3,6,12 g·kg^-1·d^-1),Chinese medicine and western medicine control groups.The chemical and physical pain threshold were observed. Nitroglycerin was used to make rat migraine model. The changes of behavior were detected. The contents of plasma NO,NOS,CGRP were measured by the way of radioactive immunoreaction or spectmphotometry. Contents of 5-HT and 5-HIAA in brain tissue were assayed HPLC. Result: Compared with the normal group, the first phase of paw licking time and the second phase of paw licking time were increased significantly in the model group of formalin test(P<0.01). In the test of migraine model induced by nitroglycerin, the forelimb scratching frequency of rats were increased markedly compared with the blank control group (P<0.01) during the period of 0 min to 30 min. The contents of NO,NOS,CGRP in model group were higher than blank group significantly(P<0.05),but the content of 5-HT and 5-HIAA in the brain was decreased (P<0.05).Xiongzhi Shigao tang could suppress Faure Marin induced pain reaction effectively, which could significantly reduce rats' first phase of paw licking time and second phase of paw licking time compared with the model group (P<0.05,P<0.01). And Xiongzhi Shigao tang could significantly increase the pain threshold of mice induced by hot plate compared with the blank control group (P<0.05,P<0.01).Compared to model group, the contents of NO,NOS and CGRP in high and middle dosage group of Xiongzhi Shigao tang were decreased (P<0.05, P<0.01), and the content of 5-HT were increased in brain (P<0.05). Conclusion: The remedial Xiongzhi Shigao tang can treat migraine through adjusting the contents of NO,NOS,CGRP and 5-HT.This might be one of the possible mechanisms of Xiongzhi Shigao tang in the treatment of migraine.