Study On The Adenosine A₁ Receptors And Signal Transduction Pathways Involved In The Mechanism Of Electroacupuncture In Ratmodels Of Migrainewith Nitroglyeerin

Objective:1. Study the behavioral impact of electro-acupuncture at different acupoint groups of migraine model rats induced by nitroglycerin(GTN), to compare the effect of different acupoints compatibility differences.2.Select the MAPK pathway and ADORA1 which is closely related to the painas the breakthrough point, to observe the analgesiceffect of electro-acupuncture and intracerebroventricular(icv.) injection of adenosine A1 receptor agonist(CCPA) and antagonist(DPCPX) on migraine model rats induced by nitroglycerin(GTN). And explore the possible role and mechanism of of adenosine A1 receptor and ERK1/2 in the MAPK pathway in the pathogenesis of migraine.At the same time, take the gene and protein expression of ERK1/2, ADOR A1 was the index, To reveal the mechanism of electroacupuncture analgesia and provide scientific experimental basis for the elucidation of the mechanism of Electro-acupuncture on migraine pain. Methods:1.Through the establishment of nitroglycerin experimental migraine rat model, to test behavioral changes of rats in each group. Then observe the treatment effect and influence of electro-acupuncture at different acupoints group on the symptoms of migraine model rats and to compare the differences between each group.2. By using methods of immunohistochemistry and real time PCR, to observe electro-acupuncture on local meridiangroup(Fengchi ? Shuaigu) and intracerebroventricular(icv.) injection of adenosine A1 receptor agonist(CCPA) and antagonist(DPCPX) on the hypothalamus and spinal cord of migraine model rats induced by nitroglycerin(GTN),andpreliminary discuss the analgesiceffect of ADOR A1 mRNA expression on the hypothalamus and spinal cord of migraine model rats induced by nitroglycerin(GTN). Byusing methods of Western Blot and real time PCR, to observe electro-acupuncture on local meridiangroup(Fengchi?Shuaigu)to the analgesiceffect of protein and gene expression of ERK1/2 in the MAPK pathwayon the hypothalamus and spinal cord of migraine model rats induced by nitroglycerin(GTN). Results:1. Behavior observation results show that: "climb cage, scratching, red eared" behavior science change can be seen as the rats pain defense reflects the objective performance can be as glyceryl trinitrate(GTN) migraine rat model behavior evaluation index. EA followed by far group(Waiguan, Yanglingquan), local acupoint group(Fengchi, the rate of the valley) were able to reduce migraine model rats behavior score, EA local acupoints group in reducing of migraine model rats behavior score is better than that of electro acupuncture followed by far fetch group.2.RT-PCR results showed that:(1) the expression of ADOR A1 mRNA in hypothalamus and spinal cord of the rat model of migraine was down regulated, and the expression of gene was up-regulated by electroacupuncture. Adora1 agonist CCPA lateral ventricle microinjection can upregulate the ADORA1 mRNA gene expression and may be coordinating role in the same direction and EA. Adora1 antagonist DPCPX side of microinjection into the ventricle inhibited the expression, and may have a reverse on electroacupuncture(EA).(2) migraine model rat hypothalamus and spinal cord ERK1 mRNA and ERK2 mRNA expression and down regulate the gene expression of electroacupuncture.3.Immunohistochemistry results showed that :ADORA1 in hypothalamus and spinal cord in rats with migraine(the expression site was in cytoplasm) protein expression, electroacupuncture can upregulate the expression of the receptor. The upregulation of the expression of protein ADORA1 activator CCPA after intracerebroventricular injection, and may have the same direction and coordination role of electroacupuncture. Inhibitory effect of ADORA1 antagonist DPCPX on the expression of micro injection in the lateral ventricle, and the inhibition effect of Electroacupuncture on electroacupuncture.4. Western Blot results showed that: migraine model rat hypothalamus and spinal cord of adenosine ERK1 and ERK2 protein expression, protein expression decreased two ea.Conclusions:1. EA followed by far fetch group, the local meridian group can obviously alleviate glyceryl trinitrate(GTN) type of migraine rats discomfort behavioral symptoms; the analgesic effect of the local meridian group is better.2. Nitroglycerin(GTN) occurrence and development of migraine may be reduced, and expression of ADORA1 in hypothalamus and spinal cord of rats ERK1/2 expression.3. EA can increase the expression of ADORA1 in the hypothalamus and spinal cord of migraine rats, and down regulate the expression of ERK1/2.