大川芎方效应组分对偏头痛大鼠降钙素基因相关肽及其受体表达的影响

Effects of active components extracted from Dachuanxiong Decoction on the expressions of CGRP and its receptors in rats with migraine

目的研究大川芎方(川芎、天麻)效应组分对偏头痛大鼠下丘脑、中脑导水管周围灰质(periaqueductal gray,PAG)中降钙素基因相关肽(calcitonin gene-related peptide,CGRP)及其受体表达的影响。方法将72只大鼠随机均分为正常组,模型组,阳性药组,大川芎方高、中、低剂量组,随机均分为A、B 2个亚组。皮下注射硝酸甘油注射液建立大鼠偏头痛模型。采用免疫组化染色法观察B组大鼠下丘脑、中脑导水管周围灰质中CGRP的表达;酶联免疫吸附测定法(enzyme linked immunosorbent assay,ELISA)测定A组大鼠血清、下丘脑及PAG中CGRP的水平;实时荧光定量PCR(real-time PCR,RT-PCR)检测A组大鼠下丘脑、PAG中CGRP及其受体CRLR、RCP、RAMP1的基因表达。结果与正常组相比,模型组血清、下丘脑、PAG中CGRP水平明显升高(P<0.01),下丘脑中CGRP、RCP表达上调(P<0.05),PAG中CGRP、CRLR表达上调,RCP表达下调(P<0.05)。给予低、中、高剂量效应组分后均可降低偏头痛大鼠血清、下丘脑、PAG中CGRP的水平,中剂量可下调下丘脑中CGRP、RCP表达(P<0.05,P<0.01),下调PAG中CGRP、CRLR表达(P<0.05),上调PAG中RCP表达(P<0.05)。结论大川芎方效应组分可能通过下调CGRP在下丘脑、PAG的表达,减少CGRP合成,抑制神经源性炎症来达到治疗偏头痛的作用。

AIM To study the effects of active components extracted from Dachuanxiong Decoction （DCXD, Chuanxiong Rhizoma, Gastrodiae Rhizoma） on the expressions of calcitonin gene-related peptide （CGRP） and its receptors in hypothalamus and periaqueductal gray （PAG） of rats with migraine. METHODS Seventy-two rats were evenly assigned to six groups at random, normal group, model group, positive drug group, groups of high- dose, middle-dose and low-dose DCXD. Each group was then divided into Subgroup A and Subgroup B. The mi- graine rat model was established by subcutaneous nitroglycerin injection. The expressions of CGRP in Subgroup B＇ s rat hypothalamus and PAG were observed by immunohistochemical staining, while the levels of CGRP in rat plas- ma, hypothalamus and PAG in Subgroup A were detected by ELISA. All the gene expressions of CGRP and its receptors （CRLR, RCP, RAMP1 ） in Subgroup A＇s rat hypothalamus and PAG were assessed by real-time PCR. RESULTS There was a more significant CGRP level increase in plasma, hypothalamus and PAG in the model group than that in the normal group （P 〈0. 01 ）. The modelled rats found their up-regulated expressions of CGRP and RCP in hypothalamus （P 〈 0. 05 ）, up-regulated expressions of CGRP and CRLR in PAG, and down-regulated expression of RCP （P 〈 0. 05 ）. All low-dose, middle-dose, high-dose active components worked in decreasing the CGRP levels in plasma, hypothalamus and PAG. The middle-dose led to down-regulation of CGRP, RCP expres- sions in hypothalamus （P 〈 0. 05, P 〈 0. 01 ）, down-regulation of CGRP, CRLR expressions in PAG （P 〈 0.05 ）, and up-regulation of RCP expression in PAG （P 〈 0. 05 ）. CONCLUSION The mechanism of active components extracted from DCXD in migraine management may associate with its capability in down-regulating CGRP expres- sions in hypothalamus and PAG, reducing CGRP synthesis and inhibiting neurogenic inflammation.