清肝祛风通络方对偏头痛模型细胞增殖及CGRP、SP蛋白表达的影响

目的探讨清肝祛风通络方(苗药)对偏头痛模型细胞的增殖及CGRP、SP蛋白表达的影响。方法原代培养大鼠大脑皮质神经细胞,硝酸甘油诱导偏头痛模型。实验分为6组:正常空白组(不造模),模型对照组(硝酸甘油处理),高、中、低剂量苗药给药组(硝酸甘油处理+苗药含药血清处理),阳性对照组(硝酸甘油处理+盐酸氟桂利嗪处理)。MTT法检测大鼠大脑皮质神经细胞存活率;Western Blot检测大鼠大脑皮质神经细胞内降钙素基因相关肽(CGRP)与P物质(SP)蛋白表达。结果模型对照组细胞存活率(55.71±2.00)%,低于正常空白组,差异有统计学意义(t=6.858,P<0.001),高、中剂量苗药给药组大鼠大脑皮质神经细胞存活率分别为(77.99±3.20)%与(70.12±2.56)%,高于模型对照组,差异有统计学意义(t=4.164,2.694,P=0.002,0.023)。CGRP、SP蛋白在正常空白组的表达最低,在模型对照组的表达最高;苗药给药组中CGRP、SP蛋白表达随苗药浓度的增加而降低,与模型对照组比较,差异有统计学意义(P<0.05或P<0.01)。结论清肝祛风通络方对硝酸甘油诱导的大鼠大脑皮质神经细胞凋亡具有明显的抑制作用,其机制可能与抑制CGRP、SP蛋白的表达有关。

Effect of Qinggan Qufeng Tongluo Recipe on Cell Proliferation and Expression of CGRP and SP Protein in Migraine Models

Objective To investigate the effects of Qinggan Qufeng Tongluo Recipe(Miaoyao) on the proliferation of migraine model cells and the expression of CGRP and SP protein. Methods primary cultured rat cortical neurons and nitroglycerin induced migraine models. The experiment was divided into six groups: normal blank group(no model), model control group(nitroglycerin treatment), high, medium and low dose Miao medicine administration group(nitroglycerin treatment+serum treatment of Miao medicine), and positive control group(nitroglycerin treatment+flunarizine hydrochloride treatment). MTT assay was used to detect the survival rate of rat cerebral cortical neurons;Western blot was used to detect the expression of calcitonin gene-related peptide(CGRP) and substance P(SP) in rat cerebral cortical neurons. Results The cell survival rate of model control group was(55.71±2.00)%, lower than that of normal blank group(t=6.858, P<0.001). The survival rate of cerebral cortex neurons in high and medium dose Miao medicine group was(77.99±3.20)% and(70.12±2.56)%, respectively, higher than that of model control group, with significant difference(t=6.858, P<0.001). T=4.164, 2.694, P=0.002, 0.023). The expression of CGRP and SP protein was the lowest in the normal blank group and the highest in the model control group. The expression of CGRP and SP protein in the Miao medicine group decreased with the increase of the concentration of Miao medicine, and there was significant difference compared with the model control group(P<0.05 or P<0.01). Conclusion Qinggan Qufeng Tongluo Recipe can significantly inhibit the apoptosis of rat cerebral cortical neurons induced by nitroglycerin, and its mechanism may be related to the inhibition of the expression of CGRP and SP proteins.