| Migraine,as a common primary neurological diseases.Unilateral moderate to severe pulsation-like headache is the most common symptom,sometimes alternating on both sides.Severe cases are repetitive and lingering,often accompanied by symptoms such as photophobia,phobia,nausea,vomiting.The current pathogenesis of migraine is still not clear.The mainstream theory believes that migraine is closely related to trigeminal vascular theory,cortical diffusion inhibition,genetic factors and biochemical factors.It is believed that an important inducer of migraine is by improper adjustment of the person with physical weakness,especially when the external elements attack on the basis of physical deficiency is the major cause.The common seen factors are the head is not dried after bathing,and when the phlegm is vented,the wind and heat channel through over exerting immune system.There are various treatment options for migraine in clinical practice,including the well documented traditional Chinese medicine in the classification and statistics literature.For the most common wind-heat syndrome,the combination of disease syndrome and animal model adopts the combination of Western medicine pathological modeling factors and TCM syndrome modeling factors.On the basis of the combined animal model,the wind-heat syndrome model plus nitroglycerin-induced model has certain research value and significance.Gastrodin and puerarin are the main active ingredients of Chinese medicine Tianma and Pueraria,respectively;often used for vertigo,hypertension,vertebral basilar artery insufficiency,sudden deafness,and other diseases.However,the clinical application of gastrodin and puerarin is mainly seen in injections,and puerarin injection also has the risk of causing adverse reactions such as hemolysis.Oral preparations of gastrodin and puerarin have been developed on the market,and the safety is higher than that of injection.However,increasing the oral bioavailability of gastrodin and puerarin has become a research hotspot.Pharmacokinetics indicate that the combination of the two can effectively increase the respective absorption rate,reduce the clearance rate,and prolong the average residence time.Exploring the efficacy of combined application of gastrodin and puerarin in the treatment of migraine can help to provide new ideas for the clinic practice.Objective:In this study,according to the disease-binding model,the model of wind-heat syndrome migraine was induced by swimming,blown by wind and then injected with nitroglycerin.The effects of gastrodin combined with puerarin on the pain threshold of wind-heat model migraine rats were explored.Whether it is more effective in suppressing pain sensitization during migraine attacks.The expression of CGRP,RAMP1 and CRLR in rat brain were used as an indicator to measure the therapeutic effect of combined application,and provide more ideas for clinical treatment of migraine.Methods:Sixty SPF male rats,weighing 180-220 g each,were randomly divided into normal control group,migraine wind-heat model group,gastrodin group,puerarin group,gastrodin combined with puerarin group.Sumatriptan group,10 in each group.Normal control group:normal feeding on the first day and the second day of the experiment,without any treatment,the third day of the experiment began to be administered with 10ml/kg/d normal saline for three consecutive days;migraine wind-heat model group:the first day of the experiment first swim,water temperature(24～26 ℃),time 30 min,after swimming,hot air in the incubator,temperature(39±0.5℃),humidity(50±5%),wind 2,for five consecutive days.On the fourth day of the experiment,the stomach was firstly administered with normal saline,10ml/kg,and then swimming and blowing.The gastrodin group:the rest of the same migraine model,the gastrodil was gastrodin(30mg/kg).Puerarin group:the rest of the treatment with the migraine wind-heat model group,the administration of puerarin(150mg/kg).On the sixth day of the experiment,2 ml/kg of normal saline was injected into the back of the control group after the gavage,and the other groups were injected with nitroglycerin 2 ml/kg.Gastrodin combined with puerarin group:the rest were treated with migraine and wind-heat model group.Gastrointestinal drugs were gastrodin and puerarin(the gastrodin 30mg/kg+puerarin 150mg/kg),and the gastric perfusion amount was 10ml/kg.Sumatriptan group:the rest of the treatment was the same as the migraine wind-heat model group.The gavage was sumatriptan,and the amount of gavage was(6mg/kg),1 ml/100 g.After 4 hours,the pain threshold of each group was measured with a rat tail light tester.Five rats in each group were perfused with fixed brain,and frozen sections were used to detect the CGRP content in rat brain slices.The other 5 rats in each group were directly taken from the thalamus.The trigeminal ganglion and the trigeminal nucleus were frozen in liquid nitrogen.The content of RAMP 1 and CRLR in rats were detected by Western-blot method.Research result:1.Observing the behavior of rats after modeling,it was found that the rats in the model group of migraine with wind-heat syndrome showed irritability,standing in cage,conjunctival congestion,red tongue,no soft stool,dry and hard texture,more times of scratching the head,red ears and more times of climbing cages.2 Pain threshold:After the model establishment,the pain threshold of the model group was significantly lower than that of the normal control group,the difference was statistically significant(P<0.01).Compared with the model group,the gastrodin group and the puerarin group were used alone.The effect was not obvious,but the effect of gastrodin combined with puerarin group was better(P<0.01),which was equivalent to that of sumatriptan group.3 Immunohistochemistry:The results of CGRP protein content in the brain slices of each group showed that the CGRP protein content in the amygdala model group was significantly higher than that in the normal control group(P<0.01);the treatment group had a lower trend than the model group.(P<0.05)The concentration of gastrodin combined with puerarin group was more significant than that of gastrodin group and puerarin group(P<0.05).The effect was similar to that of western medicine sumatriptan group.There was no statistical difference between them.4 Western-Blot:In the thalamus,there was no significant difference in RAMP1 protein expression between the migraine model and the normal group(p=0.078),but there was a significant increase in expression,and CRLR also increased significantly,with statistically significant differences.(P<0.01);There was no statistically significant difference between RAMP1 and CRLR in the combination of gastrodin and puerarin.The CRLR of gastrodin combined with puerarin group decreased significantly compared with the model group(P<0.01),and RAMP1 also decreased.The trend(p=0.09);the gastrodin combined with the puerarin group was lower than the puerarin group RAMP1 and CRLR(p<0.05);the trigeminal ganglion site,the migraine wind-heat model group,the RAMP1 and CRLR content were higher than the normal control group.There was a significant increase(p<0.01),and there was a decrease in gastrodin combined with puerarin group,among which RAMP1 was more significant(p<0.01).The content of RAMP1 and CRLR in the sumatriptan group was also lower than that in the model group(p<0.01);The content of RAMP 1 in the combination of gastrodin and puerarin was less than that in the combination of gastrodin and puerarin(p<0.05).There was no significant difference between the components in the nucleus of the trigeminal nucleus.The expressions of RAMP1(p=0.09)and CRLR(p=0.055)were increased in the migraine model group compared with the normal control group;the RAMP1 and CRLR protein contents were decreased in each treatment group compared with the model group;gastrodin combined with Pueraria The down-regulation trend of the puerarin group was higher than that of the gastrodin.Conclusion:Wind is an important cause of migraine.Wind-heat syndrome is a clinically high-rising syndrome of migraine.An important condition for the occurrence of migraine is caused by inappropriate adjustment of people with physical weakness.This experiment uses normal temperature swimming and blowing hot air.Simulating the climate characteristics of wind-heat syndrome,the classical nitroglycerin induced pathological changes of migraine,and the syndrome combined with the wind-heat syndrome migraine model.The model results showed that the model group showed redness,irritability,dryness and other similar human wind and heat symptoms,as well as migraine-like manifestations such as taro and cage.The model of the model of wind-heat syndrome migraine was more consistent with expectations..CGRP is a core participant in the pathogenesis of migraine,and its increased release is considered to be an important marker of migraine attack caused by trigeminal vascular reflex activation.The combined expression of CRLR and RAMP1 constitutes a CGRP functional receptor.In this experiment,CGRP and its receptors ramp1 and CRLR were used as models to evaluate the migraine model of wind-heat syndrome and the intervention of traditional Chinese medicine.Immunohistochemical results showed that CGRP positive expression was most obvious in the amygdala region,suggesting that CGRP can promote pain in the amygdala region,indicating that the amygdala not only participates in mood regulation,but also involves the regulation of pain and pain emotions;Westren-blot found that there was a correlation between the expression of RAMP 1 and CRLR protein in the thalamus,trigeminal ganglion,and trigeminal nucleus.Gegen relies on muscle spasm,heat,main wind,Tianma calming liver to stop endogenous wind,main sputum internal wind,the two are compatible,from the inside and outside the wind and evil.In this experiment,puerarin tablets were combined with gastrodin tablets.The results showed that the combination of drugs could significantly increase the pain threshold of the rats with migraine in the wind-heat syndrome group,and had significant analgesic effect on migraine rats;The expression of CGRP in the amygdala of rats with heat syndrome migraine model was better than that of single use;and it could lower the CGRP receptor rampl and CRLR protein in the thalamus,trigeminal ganglia and trigeminal nucleus.The expression of the content,wherein the difference in the trigeminal ganglion is most obvious.Therefore,swimming and blowing plus nitroglycerin in the back of the neck can effectively induce the behavioral characteristics of the rat model of wind-heat syndrome migraine.Puerarin tablets combined with Tianmasu tablets have obvious analgesic effect on the model rats,and can regulate CGRP and the expression of its receptors ramp1 and CRLR is involved in anti-migraine effects. |
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