天麻制剂通过腺苷途径治疗偏头痛相关的分子机制研究

Title:The molecular mechanism of the Tianma preparation in the treating migraine by adenosine pathway

目的探讨天麻制剂各有效成分对偏头痛模型大鼠的降钙素基因相关肽(calcitonin gene-related peptide,CGRP)与腺苷A1受体(adenosine A1 receptor,A1R)表达的影响。方法将SPF级雄性SD大鼠84只随机分为7组(n=12):假手术组(A组/阴性对照组)、电刺激三叉神经节(electrical stimulation of the trigeminal ganglion,ESTG)模型组(B组)、舒马普坦干预组(C组/阳性对照组)、天麻素干预组(D组)、对羟基苯甲醇干预组(E组)、香英兰醇干预组(F组)、β-谷甾醇干预组(G组)。通过建立ESTG模型,采用酶联免疫吸附测定(enzyme-linked immuno sorbent assay,Elisa)、免疫荧光及Western-Blot技术检测天麻制剂各有效成分对CGRP与A1R表达的影响。结果与A组相比,B组大鼠三叉神经节(trigeminal ganglia,TG)、三叉神经脊束尾核(trigeminal nucleus caudalis,TNC)中的CGRP表达明显增高,A1R的表达明显降低,差异具有统计学意义(P<0.01)。与B组相比,C、D组大鼠TG、TNC中的CGRP表达明显降低,A1R的表达明显增高,差异具有统计学意义(P<0.01);而E、F、G组与B组之间差异无统计学意义(P>0.05)。与C组相比,D组大鼠TG、TNC中的CGRP、A1R的表达无明显差异(P>0.05)。结论预防应用天麻素,与舒马普坦一样可对偏头痛发作起到一定保护作用,而香英兰醇、对羟基苯甲醇及β-谷甾醇对缓解偏头痛的作用疗效甚微,且天麻制剂有效成分中天麻素可通过激活A1R及抑制CGRP表达来抑制偏头痛的发生。

Objective To investigate the effect of the active ingredients of Tianma preparation on the expression of calcitonin gene-related peptide(CGRP)and adenosine A1 receptor in migraine model rats.Methods Eighty-four SPF rats were randomly divided into 7 groups(n=12):sham operation group(group A),electrical stimulation of the trigeminal ganglion group(ESTG group),Sumatriptan group(group C),gastrodin group(group D),4-hydroxybenzyl alcohol group(group E),vanilanol group(group F),β-sitosterol group(group G).The enzyme-linked immuno sorbent assay(ELISA),immunofluorescence and western-blot techniques were used to dected the effect of the active components of Tianma preparation on the expression of CGRP and adenosine A1 receptor.Results Compared with group A,the expression of CGRP in trigeminal ganglia(TG)and trigeminal nucleus caudalis(TNC)in group B rats was significantly increased,while the expression of adenosine A1 receptor was significantly decreased,with statistically significant differences(P<0.01).Compared with group B,the expression of CGRP in TG and TNC in group C and D was significantly decreased,and the expression of adenosine A1 receptor was significantly increased,with statistically significant differences(P<0.01).There was no significant difference between group E,F,G and group B(P>0.05).Compared with group C,the expression of CGRP and adenosine A1 receptor in TG and TNC of rats in group D was not statistically significant(P>0.05).Conclusion Like sumatriptan,The preventive application of gastrodin can alleviate the migraine attack,while the effects of vanillin,4-hydroxybenzyl alcoholandβ-sitosterol were not good enough.In addition,gastrodin can inhibit the occurrence of migraine by activating adenosine A1 receptor and inhibiting the release of CGRP.