散偏汤对偏头痛模型大鼠中脑、三叉神经节CGRP、PENK基因、蛋白表达的影响

Effect of Sanpian Decoction on CGRP,PENK Gene and Protein Expression in Mesencephalon and Trigeminal Ganglion of Migraine Model Rats

目的观察散偏汤对偏头痛模型大鼠中脑、三叉神经节降钙素基因相关肽(CGRP)、PENK(PENK)基因、蛋白表达的影响。方法 24只健康SPF级SD大鼠随机分为4组:生理盐水组、偏头痛模型组、琥珀酸舒马普坦组、散偏汤组。颈背部皮下注射硝酸甘油连续3周复制实验性偏头痛动物模型,第2次注射硝酸甘油后给予药物干预,1周后,第3次注射硝酸甘油2 h后取材,Western blotting法测定三叉神经节CGRP、PENK蛋白表达变化;实时聚合酶链反应PCR(Real-time PCR)检测各组大鼠中脑CGRP、PENK基因表达变化。结果 Western blotting法检测:偏头痛模型组大鼠三叉神经节中CGRP蛋白表达与生理盐水组差异有统计学意义(P<0.05),与琥珀酸舒马普坦组和散偏汤组差异有统计学意义(P<0.05),舒马普坦组与散偏汤组差异有统计学意义(P>0.05);偏头痛模型组三叉神经节PENK蛋白表达较生理盐水组降低(P<0.05),散偏汤组和琥珀酸舒马普坦组干预后PENK蛋白表达升高,与模型组差异有统计学意义(P<0.05);Real-Time PCR检测:偏头痛模型组大鼠中脑CGRP基因表达与生理盐水组差异有统计学意义(P<0.05);与琥珀酸舒马普坦组和散偏汤组差异有统计学意义(P<0.05);偏头痛模型组中脑PENK蛋白表达较生理盐水组降低(P>0.05),散偏汤组和琥珀酸舒马普坦组中脑PENK基因表达与模型组比较升高(P<0.05)。结论散偏汤可以抑制伤害性感受传导通路中CGRP的基因及蛋白表达,升高三叉神经节中PENK蛋白的表达,对偏头痛大鼠CGRP基因及蛋白的表达起抑制作用,能抑制痛觉信息的传递,并对内啡肽系统的镇痛作用有一定影响。

Objective： To study the effect of Sanpian Decoction on on CGRP,PENK gene and protein expres- sion in mesencephalon and trigemina] ganglion of migraine model rats. Methods： 24 healthy SPF grade SD rats were randomly divided into 4 groups： the saline group, the migraine model group, the sumatriptan group and San- pian Decoction group. The experimental migraine animal model was replicated with subcutaneous injection of ni- troglycerin on the back of the neck for 3 weeks. After the second injection of nitroglycerin, drug intervention was given. After 1 weeks of drug intervention,the samples were taken 2 hours after the 3rd injection,and the expres- sion of CGRP and PENK protein in the trigeminal ganglia was detected with Western-blot method. Real-time PCR was used to detect the expression of CGRP and PENK genes in midbrain of rats. Results： Western blotting method ： the expression of CGRP protein in trigeminal ganglion of migraine model group was significantly different from that in saline group （P 〈 0.05）. The difference was statistically significant with the sumatriptan succinate group and Sanpian Decoction group（P〈 0.05）. There was no significant difference between the sumatriptan group and Sanpian Decoction group （P〉 0.05 ）. The expression of PENK protein in trigeminal ganglion of migraine model group was lower than that in saline group （P〈 0.05）. The expression of PENK protein was significantly increased in Sanpian Decoction group and sumatriptan succinate group compared with the model group after intervention, which was significantly different from that in the model group （P〈 0.05 ）. Real-time PCR detection： The expres-sion of CGRP gene in the midbrain of migraine model group was significantly different from that in saline group （P〈0.05）. The difference was statistically significant with the sumatriptan succinate group and Sanpian Decoction group （P〈 0.05）. The expression of PENK protein in the midbrain of migraine model group was lower than that in normal saline group （P〈 0.05）. The expression of PENK gene in the middle cerebral cortex of Sanpian Decoction group and sumatriptan succinate group was higher than that of the model group （P〈 0.05）. Conclusion： Sanpian Decoction can inhibit the gene and protein expression of CGRP in the nociceptive transmission pathway,increase the expression of PENK protein in the trigeminal ganglia,inhibit the expression of CGRP gene and protein in the migraine rat,inhibit the transmission of pain information,and have some influence on the analgesic effect of en- dorphin system.