芎麻汤药效部位对硝酸甘油诱导偏头痛大鼠三叉神经颈髓复合体内CGRP-CRLR/RAMP1信号通路的影响

摘要

目的 用硝酸甘油型偏头痛大鼠模型,观察芎麻汤药效部位(乙酸乙酯及正丁醇提取部位)对偏头痛发作时三叉神经颈髓复合体(TCC)内降钙素基因相关肽-降钙素受体样受体-活性修饰蛋白受体1(CGRP-CRLR/RAMP1)信号通路的干预作用.方法 36只雄性SD大鼠随机分为6组(空白对照组,模型组,芎麻汤药效部位低、中、高剂量组,琥珀酸舒马普坦对照组),每组6只.皮下注射硝酸甘油(10mg/kg)复制偏头痛模型,给药后乙醚麻醉,其中18只(每组3只)心脏灌注后取全脑剥离TCC部位,采用免疫组化染色检测CGRP、CRLR、RAMP1阳性表达;余18只直接取全脑剥离TCC部位并平分两份,一份组织采用qPCR检测CGRP、CRLR、RAMP1 mRNA表达,另一份组织采用Western blot检测CGRP、CRLR、RAMP1蛋白表达.结果 与空白对照组比较,偏头痛大鼠TCC内CGRP、CRLR、RAMP1阳性细胞数、mRNA及蛋白表达均明显增加,差异具有统计学意义(P＜0.05),模型复制成功;与模型组比较,芎麻汤药效部位能降低偏头痛大鼠TCC内CGRP、CRLR、RAMP1阳性细胞数,下调CGRP、CRLR、RAMP1 mRNA和蛋白表达,差异具有统计学意义(P＜0.05).结论 芎麻汤药效部位能降低偏头痛大鼠TCC内CGRP-CRLR/RAMP1信号通路的活性.%Objective To explore the effects of bioactive parts of Xiongma Decoction (parts of ethyl acetate and n-butanol extract) on the CGRP-CRLR/RAMP1 signal pathway so as to clarify its therapeutic mechanism on migraine.Methods We randomly divided 36 male SD rats into 6 groups with 6 in each:blank group,model group,groups of low-,medium-and high-dose Xiongma Decoction bioactive parts,and Sumatriptan group.By giving hypodermic injection of 10 mg/kg nitroglycerin,the migraine rat model was copied;Only 18 rats were found to have positive expressions of CGRP,CRLR,and RAMP1 in TCC with immunohistochemistry staining after heart perfusion.For the remaining 18 rats,TCC was stripped directly from the whole brain and divided into two parts,one part used to detect CGRP,CRLR,RAMP1 mRNA expressions by qPCR,and the other part to detect CGRP,CRLR,RAMP1 protein expressions by Western blot.Results The number of CGRP,CRLR and RAMP1 immunoreactive cells,the mRNA and protein expressions on TCC in model group were effectively increased,compared with those in the blank group (P＜0.05),indicating that the model copying was successful.Compared with those in the model group,the number of CGRP,CRLR and RAMP1 immunoreactive cells in Xiongma Decoction bioactive parts was significantlv decreased,and the expressions of CGRP,CRLR and RAMP1 mRNA and protein were reduced (P＜0.05).Conclusion The bioactive parts of Xiongma Decoction can reduce the activity of CGRP-CRLR/RAMP1 signal pathway in TCC of migraine rats.