解郁止痛方对偏头痛-抑郁病模大鼠行为学及c-fos、c-jun基因表达影响

Effect of Jieyu Zhitong Decoction(解郁止痛方)on Behavior and c-fos and c-jun Gene Expression in Migraine-Depression Model Rats

目的观察解郁止痛方对偏头痛-抑郁病模大鼠行为学及c-fos、c-jun基因表达的影响。方法将72只雄性SD大鼠随机分为空白组、模型组、尼莫地平组以及解郁止痛高、中、低剂量组,采用间歇皮下注射硝酸甘油注射液(10 mg/kg)与慢性不可预知温和刺激复制偏头痛-抑郁大鼠模型,造模同时给予干预,空白组及模型组灌服等量的纯水,尼莫地平组予尼莫地平混悬液6 mg/kg灌胃,解郁止痛方高、中、低剂量组分别给予解郁止痛方混悬液(1.0 g/100 g,0.5 g/100 g,0.25 g/100 g生药)灌胃,各组干预1次/d,共21 d。观察记录造模干预各阶段各组大鼠体质量、旷场试验、强迫游泳实验、糖水消耗实验结果以及大鼠偏头痛发作时挠头、攀笼次数。第21天心脏灌注处死大鼠后取脑,用免疫组化方法记录大鼠杏仁核c-fos、c-jun基因表达。结果造模干预中(第10天)及造模干预结束时(第21天),与空白组相比,其他各组大鼠体质量均明显减轻(P<0.01,P<0.05);模型组大鼠体质量下降最明显;与模型组相比,尼莫地平组、解郁止痛方各剂量组体质量均升高(P<0.01,P<0.05)。造模干预中(第10天)及造模干预结束时(第21天),与空白组相比,其他各组水平及垂直活动得分均下降(P<0.01,P<0.05);与模型组相比,尼莫地平组、解郁止痛方各剂量组水平及垂直活动得分均显著升高(均P<0.01)。造模干预中(第10天)及造模干预结束时(第21天),与空白组相比,其他各组大鼠游泳不动时间延长(P<0.01,P<0.05);与模型组相比,解郁止痛方各剂量组、尼莫地平组游泳不动时间显著降低(P<0.01,P<0.05),解郁止痛方各剂量组与尼莫地平组相比,差异无统计学意义(P>0.05)。造模干预中(第10天)及造模干预结束时(第21天),与模型组相比,解郁止痛方各剂量组、尼莫地平组糖水偏好百分率均显著升高(均P<0.01),解郁止痛方各剂量组与尼莫地平组相比,差异无统计学意义(P>0.05)。造模干预第7天,与模型组比较,尼莫地平组、解郁止痛方中、高剂量组挠头次数减少(P<0.01,P<0.05),尼莫地平组、解郁止痛方各剂量组攀笼次数减少(P<0.01,P<0.05);造模干预第14天,与模型组比较,尼莫地平组、解郁止痛方各剂量组搔头及攀笼次数均显著减少(P<0.01);造模干预第21天,与模型组比较,尼莫地平组、解郁止痛方各剂量组搔头及攀笼次数均显著减少(P<0.01),解郁止痛方不同剂量组给药效果差异无统计学意义,但从总体趋势来看高剂量组优于低、中剂量组。空白组可观察到少量的免疫阳性细胞,模型组、尼莫地平组、解郁止痛方各剂量组阳性细胞数明显增多,吸光度值明显增高,与空白组形成显著差异,P<0.01。与模型组相比,各药物治疗组c-fos、c-jun阳性细胞数减少,吸光度值降低,P<0.01,差异无统计学意义。结论解郁止痛方可以明显改善偏头痛-抑郁模型大鼠行为学症状,保护应激引起的神经元受损。

Objective To observe the effect of Jieyu Zhitong Decoction(解郁止痛方)on the behavior and the expression of c-fos and c-jun genes in migraine-depression rats.Methods Seventy-two male SD rats were randomly divided into normal group,model group,nimodipine group and Jieyu Zhitong Decoction high,medium and low group.The migraine-depression rat model was made by intermittent subcutaneous injection of nitroglycerin(10 mg/kg)and chronic unpredictable mild stimulation,and intervention was given at the same time.Normal group and model group were given the same amount of ultrapure water,nimodipine group was given nimodipine suspension 6 mg/kg by gavage,Jieyu Zhitong Decoction high,medium and low group were given Jieyu Zhitong suspension with high,medium and low doses(1.0 g/100 g,0.5 g/100 g,0.25 g/100 g)once a day.To observe the weight,open field test,forced swimming test,sugar water consumption test results of rats in each stage of modeling intervention,and the times of scratching the head and climbing the cage during migraine.On the 21st day,the brain was harvested by heart perfusion,and the expression of c-fos and c-jun genes in rat amygdala was recorded by immunohistochemistry.Results During the intervention(day 10)and at the end of the intervention(day 21),compared with the blank group,the body mass of rats in other groups was significantly reduced(P<0.01,P<0.05);The body mass of the model group decreased most obviously;Compared with the model group,the body mass of the nimodipine group and the Jieyu Zhitong Decoction groups increased(P<0.01,P<0.05).During the intervention(day 10)and at the end of the intervention(day 21),compared with the blank group,the scores of horizontal and vertical activities in other groups decreased(P<0.01,P<0.05);Compared with the model group,the horizontal and vertical activity scores of nimodipine group and Jieyu Zhitong Decoction groups were significantly higher(all P<0.01).During the intervention(day 10)and at the end of the intervention(day 21),compared with the blank group,the swimming immobility time of rats in other groups was prolonged(P<0.01,P<0.05);Compared with the model group,the swimming immobility time of each dose group of Jieyu Zhitong Decoction and nimodipine group was significantly decreased(P<0.01,P<0.05).There was no significant difference between each dose group of Jieyu Zhitong Decoction and nimodipine group(P>0.05).During the modeling intervention(day 10)and at the end of the modeling intervention(day 21),compared with the model group,the sugar water preference percentage of each dose group of Jieyu Zhitong Decoction and nimodipine group was significantly increased(all P<0.01),and there was no significant difference between each dose group of Jieyu Zhitong Decoction and nimodipine group(P>0.05).On the 7th day of model intervention,compared with the model group,the number of head scratching in the nimodipine group and the middle and high dose groups of Jieyu Zhitong Decoction decreased(P<0.01,P<0.05),and the number of cage climbing in the nimodipine group and the Jieyu Zhitong Decoction groups decreased(P<0.01,P<0.05);On the 14th day of model intervention,compared with the model group,the times of scratching and climbing cages in Nimodipine group and Jieyu Zhitong Decoction groups were significantly reduced(P<0.01);On the 21st day of model intervention,compared with the model group,the times of scratching and climbing cages in the nimodipine group and the Jieyu Zhitong Decoction groups were significantly reduced(P<0.01).There was no significant difference in the administration effect of the Jieyu Zhitong Decoction groups,but from the overall trend,the high dose group was better than the low and medium dose groups.A small number of immune positive cells were observed in the blank group.The number of positive cells in the model group,the nimodipine group and the Jieyu Zhitong Decoction groups were significantly increased,and the optical density value was significantly increased,which was significantly different from the blank group(P<0.01).Compared with the model group,the number of c-fos and c-jun positive cells in each drug treatment group decreased,and the optical density value decreased(P<0.01),and the difference was not statistically significant.Conclusion Jieyu Zhitong Decoction can obviously improve behavioral symptoms of migrainedepression model rats and protect neurons from damage caused by stress.