解郁止痛方对偏头痛—抑郁共病模型大鼠行为学及脑组织中Iba1/RhoA蛋白表达的影响

Effects of antidepressant and analgesic formula on behavior and expression of Iba1/RhoA protein in brain tissue of rats with migraine-depression co-morbidity model

关键词:  偏头痛—抑郁  小胶质细胞  解郁止痛方  Iba1  RhoA  中枢敏化

Key words:  migraine-depression  microglia  antidepressant and analgesic formula  Iba1  RhoA  central sensitization

摘要:

[目的] 观察解郁止痛方对偏头痛-抑郁共病大鼠行为学及脑组织中电离钙结合适应分子1(Iba1),神经再生抑制因子Ras同源物基因A(RhoA)蛋白表达的影响,探讨其缓解偏头痛伴抑郁症状的分子机制。[方法] 将54只健康雄性SD大鼠随机分为空白组、模型组、西药组、中药解郁止痛方低、中、高剂量组。除空白组外,其余5组均予以颈部皮下注射硝酸甘油注射液及慢性不可预知性刺激以建立偏头痛-抑郁共病模型大鼠。实验开始造模同时予以药物灌胃干预。复制偏头痛模型前进行机械痛阈值的测定,实验造模治疗前、造模第10天、造模治疗后分别进行行为学评估(包括各组大鼠体质量、旷场实验、糖水消耗、强迫游泳实验、新奇抑食实验)。造模治疗结束后,灌注取脑,用蛋白印迹法检测大鼠脑组织杏仁核中Iba1及RhoA基因蛋白的表达。[结果] 与空白组比较,模型组大鼠体质量、旷场实验水平和垂直运动评分、糖水偏好率、机械痛阈值均下降(P＜0.01),新奇抑食-潜伏时间及强迫游泳不动时间延长(P＜0.01);与模型组比较,西药及解郁止痛方各剂量组大鼠体质量、旷场实验水平和垂直运动评分、糖水偏好率、机械痛阈值均升高(P＜0.05或P＜0.01),新奇抑食-潜伏时间及强迫游泳不动时间缩短(P＜0.05或P＜0.01)。蛋白印迹法结果显示:大鼠脑组织杏仁核中Iba1蛋白表达,与空白组比较,模型组升高;与模型组比较,西药及中药各剂量组下降,但均无统计学意义(P＞0.05)。大鼠脑组织杏仁核中RhoA蛋白表达,与空白组相比,模型组显著升高(P＜0.01);与模型组相比,西药组及中药高剂量组显著降低(P＜0.01)。[结论] 解郁止痛方可以明显改善偏头痛-抑郁共病大鼠的症状,其作用机制可能与减少中枢敏化、抑制小胶质细胞激活及影响RhoA表达有关。

Abstract:

[Objective] To observe the effects of the antidepressant formula on the behavior and expression of ionized calcium binding adaptor molecule 1 (Ionized calcium binding adaptor molecule-1,Iba1) and neurodegenerative inhibitory factor Ras homolog A (Ras homolog gene family,member A,RhoA) protein in brain tissues of migraine-depressed rats,and to explore the molecular mechanism of its alleviation of migraine with depressive symptoms.[Methods] Fifty-four healthy male SD rats were randomly divided into blank group,model group,western medicine group,and low-dose,medium-dose,and high-dose groups of Chinese herbal medicine for relieving depression and pain. After 1 week of acclimatization feeding,all 5 groups except the blank group were given subcutaneous injection of nitroglycerin injection into the neck and chronic unpredictable stimulation to establish migraine-depression co-model rats. The experimental modeling was started with pharmacological gavage intervention. Mechanical pain thresholds were measured before replication of the migraine model,and behavioral assessments (including body weight,absenteeism,sugar water consumption,forced swimming,and novelty food suppression) were performed before,on day 10,and after the modeling treatment. At the end of the modelling treatment,the brain was extracted by perfusion and the expression of Iba1 and RhoA gene proteins in the amygdala of rat brain tissue was detected by protein blotting.[Results] Compared with the blank group,the rats in the model group showed a decrease in body weight,the level and vertical movement scores of the open-field experiment,the sugar-water preference rate,and the mechanical pain threshold (P＜0.01),and an increase in the novel food suppression-latency time and forced swimming immobility time (P＜0.01);compared with the model group,the rats in the western medicine and antidepressant formula dose groups showed an increase in body weight,the level and vertical movement scores of the open-field experiment,the sugar-water preference rate,and the mechanical pain threshold (P＜0.05,P＜0.01). were increased (P＜0.05,P＜0.01),and the novel food suppression-latency time and forced swimming immobility time were shortened (P＜0.05,P＜0.01). The results of protein blotting showed that Iba1 protein expression in the amygdala of rat brain tissue was elevated in the model group compared with the blank group,and decreased in each dose group of Western medicine and traditional Chinese medicine (TCM) compared with the model group,but none of them was statistically significant (P＞0.05). The expression of RhoA protein in the amygdala of rat brain tissue was significantly higher in the model group compared with the blank group (P＜0.01);compared with the model group,it was significantly lower in the Western medicine group and the high-dose group of TCM (P＜0.01).[Conclusion] The mechanism of action of the antidepressant formula,which can significantly improve the symptoms of migraine-depression co-morbidities in rats,may be related to the reduction of central sensitization,the inhibition of microglia activation and the influence of RhoA expression.