P型Na+-K+-ATP酶与偏头痛关系的实验研究

目的 利用硝酸甘油实验性偏头痛模型探讨P型Na+-K+-ATP酶在偏头痛发病机制中的作用.方法 20只SD大鼠(雌雄各半)随机分为生理盐水对照组和模型组.模型组按TassorelliCristina法复制偏头痛大鼠模型(每周1次,连续4次),第4次造模后分别取三叉神经节及三叉颈复合体、大脑额叶组织;RT-PCR及Western印迹法分别测定P型Na+-K+-ATP酶的mRNA和蛋白的表达量,免疫沉淀法测定其活性,同时用Fluo-3/AM荧光法测定细胞内钙离子浓度.结果 模型组大鼠三叉神经节及三叉颈复合体(0.72±0.05,0.59±0.05,5.21±0.51)及大脑额叶组织(0.70±0.05,0.60±0.05,3.61±0.49)的P型Na泵mRNA及蛋白表达和活性均明显低于生理盐水对照组(0.83±0.10,0.67±0.06,6.53±0.73;0.81±0.08,0.71±0.09,6.61±0.73),差异有统计学意义;模型组大鼠三叉神经节及三叉颈复合体(211 182±12 973)及大脑额叶组织(186 511±18 297)的细胞内钙离子浓度均明显高于生理盐水对照组(135 243±18 105,143 289±25 175),差异有统计学意义.结论 P型Na+-K+-ATP酶可能通过其表达量及活性的变化参与了偏头痛的发生发展过程.

Relationship between P-type Na+ -K+ -Atpase and migraine: an experimental study with rats

Objective To investigate the role of P-type Na+ -K+ -ATPase in the mechanism of migraine. Methods nitroglycerin induced migraine Twenty Sprague-Dawley rats, 10 male and 10 female,were randomly divided into 2 groups: model group, undergoing subcutaneous injection of nitroglycerin 10 mg/kg once a week for 4 weeks so as to establish migraine model, and control group, undergoing subcutaneous injection of normal saline. Then the rats were killed with their trigeminal ganglia,trigeminocervical complex, and cortex of frontal lobe taken out. RT-PCR and Western-blotting were used to detect the mRNA and protein expression of P-type Na+ -K+ -ATPase. Immuno-precipitation assay was conducted to observe the Na + -K + -ATPase activity. The concentration of intracellular Ca2 + was investigated by Fluo-3/AM fluorometric method. Results The mRNA and protein expression levels and activity of P-type Na + -K + -ATPase in the trigeminal ganglion and trigeminocervical complex of the model group were 0. 72 ±0. 05,0. 59 ± 0. 05 ,and 5.21 ± 0. 51 respectively, and the mRNA and protein expression levels and activity of P-type Na + -K + -ATPase in the cortex of frontal lobe of the model group were 0. 70 ± 0. 05,0. 60 ± 0. 05,and 3.61 ± 0. 49 respectively, all significantly lower than those of the control group (0. 83 ± 0. 10,0. 67 ±0.06,6.53 ±0.73,0.81 ±0.08,0.71 ± 0.09, and 6.61 ±0.73 respectively, all P ＜0.05). The concentration of intracellular Ca2+ in trigeminal ganglion and trigeminocervical complex and the concentration of Ca2+ in the cortex of frontal lobe of the model group were 211 182 ± 12 973 and 186 511 ± 18 297respectively, both significantly higher than those of the control group ( 135 243 ± 18 105 and 143 289 ±25 175 respectively, both P ＜ O. 01 ). Conclusion P-type Na + -K + -ATPase may infect the pathogenesis of migraine by its expression and activity.