正天丸对偏头痛模型大鼠三叉神经二级神经元P2X_3受体表达的影响研究

Effect of Zhengtian Pills on P2X_3 Receptor Expression in Secondary Neurons of Trigeminal Nerve of Rat Model of Migraine

目的探究正天丸对偏头痛模型大鼠三叉神经二级神经元P2X3受体表达的影响。方法 2014年9—10月选取SPF级SD雄性大鼠30只,采用随机数字表法分为3组,分别为对照组、模型组、正天丸组,每组10只。正天丸组大鼠给予1.62 g·kg^(-1)·d^(-1)正天丸灌胃,对照组和模型组大鼠给予1 ml 0.9%氯化钠溶液灌胃,各组大鼠连续给药7 d。模型组、正天丸组大鼠末次灌胃30 min后左侧肩部皮下注射10 mg/kg硝酸甘油注射液,制备实验性偏头痛动物模型,对照组大鼠同期左侧肩部皮下注射0.5 ml 0.9%氯化钠溶液。观察各组大鼠耳红出现、消失时间及挠头开始、结束时间等行为学表现,采用免疫荧光、Western blotting法、实时荧光定量PCR法检测各组大鼠三叉神经颈髓复合体(TCC)中P2X_3受体及其mRNA表达水平。结果模型组和正天丸组大鼠耳红出现时间、挠头开始时间比较,差异无统计学意义(P>0.05);正天丸组大鼠耳红消失时间和挠头结束时间均较模型组提前(P<0.05)。免疫荧光结果显示,对照组、模型组和正天丸组大鼠三叉神经TCC中均有P2X_3受体阳性表达,主要分布在三叉神经脊束核尾部(TNC)和C_1、C_2颈髓背侧核浅层(Ⅰ、Ⅱ层),模型组表达亮度较对照组升高,正天丸组表达亮度较模型组降低。模型组和正天丸组大鼠三叉神经TCC中P2X_3受体及其mRNA表达水平较对照组升高(P<0.05);正天丸组大鼠三叉神经TCC中P2X_3受体及其mRNA表达水平较模型组降低(P<0.05)。结论正天丸对偏头痛的治疗作用可能与其可以抑制三叉神经二级神经元P2X_3受体表达上调有关。

Objective To investigate the effect of Zhengtian pills on P2X3 receptor expression in secondary neurons of trigeminal nerve of rat model of migraine. Methods From September to October in 2014,we selected 30 SPF grade Sprague -Dawle（SD）male rats and using the random number table method divided them into three groups：control group,model group and Zhengtian pills group,with 10 rats in each group. Rats of Zhengtian pills group was given 1. 62 g·kg - 1 ·d - 1 Zhengtian pills by gavage,rats of control group and model group were given 1 m 0. 9% sodium chloride solution by gavage. The drug administration lasted for 7 days. In model group and Zhengtian pills group,migraine rat models were established by subcutaneous injection of nitroglycerin（10 mg/ kg）at the left shoulder at 30 min after the last drug administration. Rats of control group were given 0. 5 ml 0. 9% sodium chloride solution by subcutaneous injection at the left shoulder. Behavioural performances of the rats were observed,including the appearing and disappearing time of red ear and the beginning and ending time of scratching head. Immunoflourescence,Western blotting and Real - time PCR were used to detect the expressions of P2X3 receptor and its mRNA in trigeminocervical complex（TCC）. Results Model group and Zhengtian pills group were not significantly different in＆amp;nbsp;the appearing time of red ear and the beginning time of scratching head（ P 〉 0. 05 ） . Zhengtian pills group had earlier disappearing time of red ear and earlier ending time of scratching head than model group（ P 〈 0. 05）. Immunofluorescence results showed that the expression of P2X3 receptors in TCC was positive in the rats of each group,mainly distributed in the TNC andⅠ,Ⅱ layer of C1 ,C2 cervical spinal dorsal nucleus,and the expression of model group was higher than control group and Zhengtian pills group. The expression levels of P2X3 receptor and its mRNA in TCC of model group and Zhengtian pills group were higher than those of control group（P 〈 0. 05）,while the expression levels of P2X3 receptor and its mRNA of Zhengtian pills group were lower than those of model group（P 〈 0. 05）. Conclusion The effect of Zhengtian pills on the treatment of migraine may be related with its inhibition on the expression of P2X3 receptors in secondary neurons of trigeminal nerve.