解郁止痛方对偏头痛-抑郁共病模型大鼠行为学及NLRP3和Caspase-1基因表达的影响

Effects of Jieyu Zhitong Formula on behaviors and expressions of NLRP3 and Caspase-1 genes in migraine-depression model rats

目的探究解郁止痛方(JYZT)对偏头痛-抑郁共病模型大鼠行为学及脑组织中Nod样受体蛋白3(Nod-like receptor protein 3,NLRP3)/胱天蛋白酶-1(Caspase-1)蛋白表达水平的影响。方法48只大鼠随机分为正常组、模型组、阳性药物组、JYZT低剂量组、JYZT中剂量组、JYZT高剂量组,每组8只,除正常组外,其余各组均采用间歇性颈部皮下注射硝酸甘油注射液及7种不同的慢性不可预知性温和刺激方法诱导偏头痛-抑郁共病大鼠模型。造模开始同时予以药物干预治疗。诱导偏头痛模型前进行热痛阈值测定,造模前、造模治疗中(第10天)、造模治疗后对各组大鼠进行行为学测试。Western blot检测大鼠杏仁核组织中NLRP3及Caspase-1蛋白的表达。结果第7、第14、第21天,与正常组相比,模型组热痛阈值显著降低(P<0.01);与模型组相比,阳性药物组及解郁止痛方各剂量组热痛阈值显著升高(P<0.05,P<0.01)。造模治疗第10天、第21天,与正常组相比,模型组旷场实验水平和垂直运动得分、糖水偏好率均显著降低(P<0.01),新奇抑制摄食-潜伏时间及强迫游泳不动时间均显著延长(P<0.01);与模型组比较,阳性药物组及JYZT各剂量组旷场实验水平和垂直运动得分、糖水偏好率均显著升高(P<0.05,P<0.01),新奇抑制摄食-潜伏时间及强迫游泳不动时间显著缩短(P<0.05,P<0.01)。造模治疗结束后,与正常组比较,模型组NLRP3及Caspase-1蛋白表达显著升高(P<0.01);与模型组比较,阳性药物组及JYZT中剂量组、JYZT高剂量组NLRP3及Caspase-1蛋白表达显著降低(P<0.01)。结论解郁止痛方可以改善偏头痛-抑郁共病大鼠的行为学得分,降低大鼠脑组织杏仁核中NLRP3及Caspase-1蛋白表达,从而抑制炎症反应的发生。

Objective To investigate the effects of Jieyu Zhitong(JYZT)Formula,on the behavioral and expression levels of nod-like receptor protein 3 and Caspase-1 in brain tissues of migraine-depression rat model.Methods Forty-eight SD rats were randomly divided into normal,model,positive drug and JYZT low-,medium-and high-dose groups,with 8 in each group.All groups except the normal group were induced with intermittent subcutaneous neck nitroglycerin injection and 7 different chronic unpredictable mild stimulation methods for migraine-depression rat models.Pharmacological interventions were administered at the beginning of modeling.Thermal pain thresholds were measured before the induction of the migraine model,and behavioral tests were performed in each group before,during(10 d),and after the modeling.After the experiments,the expressions of NLRP3 and Caspase-1 protein in the amygdala tissue of rats was detected by Western blot.Results On the 7th,14th and 21st days,the thermal pain thresholds in the model group decreased significantly compared with the normal group(P<0.01),and the thermal pain thresholds in positive drug group and each dose group of antidepressant formula increased in comparison to the model group(P<0.05,P<0.01);on the 10th and 21st days of the modeling,compared with the normal group,the model group showed decrease in the horizontal and vertical movement scores of the model group in the open field test,and sugar water preference rates(P<0.01),and a prolongation of the novel food suppression-latency time and forced swimming immobility time(P<0.01).The positive drug group and JYZT dose groups showed an increase in the level and vertical locomotion scores and sugar-water preference rates(P<0.05,P<0.01)and a shortening of the novel food suppression-latency time and forced swimming immobility time(P<0.05,P<0.01).After modeling,the expressions of NLRP3 and Caspase-1 protein in the model group was significantly higher compared with those of the normal group(P<0.01);the expressions of NLRP3 and Caspase-1 protein in the positive drug group and JYZT medium-and high-dose groups were significantly reduced(P<0.01).Conclusion JYZT Formula,can improve the behavioral scores of migraine-depressed rats,reduce the expression of NLRP3 and Caspase-1 protein in the amygdala of rat brain tissues,and thus inhibit the inflammatory response.