中药治疗偏头痛处方规律分析及脑痛立停胶囊治疗偏头痛作用机理研究
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摘要
目的:探讨治疗偏头痛中药新药脑痛立停胶囊治疗偏头痛的短期镇痛作用机理和预防偏头痛发作的长期治疗作用机理。
方法:检索治疗偏头痛的古代方剂和现代用于治疗偏头痛的中药处方,分析中药治疗偏头痛的处方规律,为脑痛立停胶囊的组方提供理论与文献依据。建立电刺激上矢状窦(SSS)区硬脑膜大鼠偏头痛模型,给与脑痛立停胶囊灌胃治疗,并以非甾体类抗炎药萘普生为对照药物,用免疫组化SABC法检测脑干和下丘脑c-fos、c-jun表达水平,放射免疫法检测血浆降钙素基因相关肽(CGRP)、内皮素(ET)含量;建立利血平化低5-羟色胺(5-HT)小鼠偏头痛模型,给与脑痛立停胶囊灌胃治疗,以复方羊角胶囊为对照药物,采用电化学法测定小鼠血浆和脑组织5-HT、5-羟吲哚乙酸(5-HIAA)、多巴胺(DA)、高香草酸(HVA)、3,4-二羟基苯乙酸(DOPAC)和去甲肾上腺素(NE)的含量,用放射免疫法检测脑组织亮氨酸-脑啡肽(L-EK)和β-内啡呔(β-EP)的含量;采用小鼠抗焦虑实验和小鼠爬梯实验评价脑痛立停胶囊的抗焦虑作用。
结果:
1.无论现代处方还是古代方剂,祛风散寒止痛药都是使用频率最高的药物。用于治疗偏头痛的现代处方和古代方剂中,川芎、白芷和细辛都是最常用的药物。从功效方面看,用于治疗偏头痛的现代处方中,多以活血化瘀,通络止痛,祛风止痛,平肝息风为主。
2.电刺激SSS区硬脑膜偏头痛模型大鼠脑干和下丘脑c-fos、c-jun阳性细胞数及阳性细胞面积明显高于正常对照组和假手术组,经脑痛立停胶囊灌胃治疗后,模型大鼠脑干和下丘脑c-fos、c-jun阳性细胞数及阳性细胞面积明显下降,其中大剂量组作用明显,其作用强度与萘普生相当。
3.电刺激SSS区硬脑膜偏头痛模型大鼠血浆CGRP、ET含量明显升高,脑痛立停胶囊可明显降低模型动物血浆CGRP水平。但对升高的血浆ET无明显影响。4. ICR小鼠对利血平的反应较为敏感,与文献报道的KM小鼠在利血平使用剂量方面有很大差异。本实验皮下注射利血平0.25mg/kg方法复制的偏头痛小鼠模型血浆和脑组织5-HT、NE、DA含量较正常小鼠明显减少,说明模型复制成功。
5.利血平化低5-HT偏头痛模型小鼠体重下降、体温降低、凝血时间缩短,血浆及脑组织5-HT、DA的含量明显降低,脑组织NE、和HVA含量也明显降低。脑痛立停胶囊可以改善利血平化低5-HT偏头痛模型小鼠的一般状况,提高模型小鼠的体重、体温并延长模型小鼠的凝血时间;可提高模型小鼠血浆和脑组织5-HT、DA及脑组织NE的含量,同时降低模型小鼠血浆DOPAC含量,作用强度与复方羊角胶囊相似;脑痛立停胶囊对模型小鼠血浆5-HIAA和HVA含量无显著影响。
6.利血平化低5-HT偏头痛模型小鼠血浆和脑组织5-HT、DA和脑组织NE含量下降的同时,脑组织L-EK含量也明显低于正常对照组和假手术组。脑痛立停胶囊在提高利血平化低5-HT偏头痛模型小鼠血浆和脑组织5-HT、DA含量和脑组织NE的同时,能够明显提高模型小鼠脑组织L-EK和β-EP的含量。7.脑痛立停胶囊可以增加小鼠抗焦虑实验跨箱次数,缩短黑箱停留时间;同时可增加爬梯实验的爬梯级数,减少小鼠直立次数。
结论:
1.脑痛立停胶囊可能通过以下机制发挥短期缓解疼痛作用:通过某些作用机制抑制脑干、下丘脑c-fos与c-jun基因的表达,减弱或阻断了疼痛刺激在中枢神经的传导;降低血浆CGRP的含量,从而发挥调节血管舒缩障碍及抗炎作用;提高血浆及脑组织中5-HT、DA和脑组织NE的含量而起到调整脑部血管舒缩功能作用;调节血小板功能,减轻血液高凝状态,改善脑部血液循环。
2.脑痛立停胶囊可能通过以下机制发挥预防偏头痛发作的长期治疗作用:提高脑组织5-HT、NE、DA、L-EK和β-EP的含量,加强偏头痛患者中枢痛调制系统的功能;改善偏头痛患者的焦虑症状。
Objective
To discuss the action mechanism of Naotongliting capsule as a new medicine used to treat patients with migraine both on long term protective function and immediate effect of alleviating pain.
Methods
Both ancient and modern traditional Chinese medicine (TCM) prescriptions used to cure migraine are collected for finding the general rule of herb selection among those prescriptions by statistical analyzing;After making rat model with migraine by electrostimulating superior sagittal sinus(SSS),the model rats are treated whit Naotongliting capsule meanwhile taking Naproxen Sodium as positive control .The protein products of c-fos and c-jun gene in brine stem and hypothalamus of rats are tested with immunohistochemical method SABC.The content of calaitonin gene related peptide(CGRP) and endothelin(ET) in blood plasma of rats is measured by means of radioimmunity test; Mice model with migraine caused by exhausting 5-hydroxytryptamine(5-HT) with Reserpine is built up ,then the model mice are treated whit Naotongliting capsule.Fufangyangjiao capsule is used as control medicine.The content of 5-HT, 5-hydroxyindole acetic acid(5-HIAA),dopamine(DA),homovanillic acid(HVA),dihydroxyphenylacetic acid(DOPAC)and norepinephrine(NE)in brain and blood plasma of mice are tested by electrochemical meathod.The level of leucine enkephalin(L-EK) andβ- endorphin(β-EP) in brain of mice is tested with radioimmunity method;Staircase test and anti-anxiety test are used to estimate the dissipate anxiety effect of Naotongliting capsule
Results
1. The herbal medicine which have functions of dispelling wind-evil, dispersing cold and alleviating pain are most widely used not only in ancient TCM prescriptions, but also modern TCM prescriptions. Among the TCM prescriptions treating migraine.Chuanxiong, Baizhi and Xixin are used most frequently, and most of those prescriptions have efficacy virtue of stimulating circulation to end stasis, expelling the obstruction from collaterals and calming the liver to stop the wind.
2. Model rats with migraine caused by electrostimulating SSS have more cells which express protein products of c-fos and c-jun gene in brine stem and hypothalamus ,and the area of c-fos and c-jun positive cells is higher than normal control group and false operation group. Naotongliting capsule can decrease the mumber and the area of c-fos and c-jun positive cells of model rats.Big dose therapy group has better effect that is similar to Naproxen Sodium.
3. The level of CGRP and ET in blood plasma of model rats with migraine caused by electrostimulating SSS is upregulated. Naotongliting capsule can downregulate the level of CGRP in blood plasma of model rats,but it has no obvious effect on upregulated ET level .
4. According to the results of this research, ICR mouse is sensitive to Reserpine.The dose of Reserpine ICR mouse can endureis quite different to this of KM mouse that is reported in articles. In this experiment ,the dose of Reserpine giving to ICR mouse is 0.25mg/kg when causing mice model with migraine successfuly.The content of 5-HT,DA and NE in blood plasma and brain stem and hypothalamus is decreased obviously.
5. Model mice with migraine caused by exhausting 5-HT with Reserpine have low temperature and low body weight and short blood clotting time.The content of 5-HT,DA in blood plasma ,brain stem and hypothalamus of model Mice is decreased ,and NE,HVA in brine stem and hypothalamus is decreased too. Naotongliting capsule can better the general condition of model mice, increase temperature and body weight, prolong blood clotting time of model mice;Meanwhile the content of 5-HT,DA in blood plasma ,brain stem and hypothalamus of model mice are increased,and the content of DOPAC in plasma is downregulated. The effect is similar to Fufangyangjiao capsule.But Naotongliting capsule has no obvious effect on 5-HIAA and DA in plasma.
6. Model mice with migraine caused by exhausting 5-HT with Reserpine not only have lower content of 5-HT and DA in plasma, brain stem and hypothalamus, lower content of NE in brine stem and hypothalamus,but also have lower level of L-EK andβ-EP in brain than normal control and false injection groups. Naotongliting capsule can also upregulate the level of L-EK andβ-EP in brine, when upregulate the content of 5-HT and DA in plasma and brain stem and hypothalamus, upregulate the content of NE in brain of model mice.There is an obvious relation between effect and dose.
7. The Naotongliting capsule can increases frequency with which the mice stride over the light-dark chambers,and abbreviate the time of mice staying in the dark chamber;Meanwhile It can increases the number of staircases that the mice clamber,and decreases the frequency with which the mice arises.
Conclusions
1. Naotongliting capsule could give its immediate effect of alleviating pain by following mechanism:restraining the expression of c-fos and c-jun gene by some kind of way, weakening or stopping the conduction of pain stimulation in the center nerves system;Downregulating the level of CGRP in plasma to regulate the function of blood vessel and improv anti-inflammatory effect;Upregulating the level of 5-HT and DA in plasma , brain stem and hypothalamus, increasing the content of NE in brine stem and hypothalamus,to regulate function disorder of blood vessel in brain;Regulating function of thrombocyte, abating the blood clotting condition to improving the blood circulation in brain.
2. Naotongliting capsule could give its long term protective function by following mechanism: Upregulating the level of 5-HT, NE, DA, L-EK andβ-EP in brain,to improve the function of pain modulation of the center nerves system; Calming the anxiety symptoms of patients with migraine.
方法:检索治疗偏头痛的古代方剂和现代用于治疗偏头痛的中药处方,分析中药治疗偏头痛的处方规律,为脑痛立停胶囊的组方提供理论与文献依据。建立电刺激上矢状窦(SSS)区硬脑膜大鼠偏头痛模型,给与脑痛立停胶囊灌胃治疗,并以非甾体类抗炎药萘普生为对照药物,用免疫组化SABC法检测脑干和下丘脑c-fos、c-jun表达水平,放射免疫法检测血浆降钙素基因相关肽(CGRP)、内皮素(ET)含量;建立利血平化低5-羟色胺(5-HT)小鼠偏头痛模型,给与脑痛立停胶囊灌胃治疗,以复方羊角胶囊为对照药物,采用电化学法测定小鼠血浆和脑组织5-HT、5-羟吲哚乙酸(5-HIAA)、多巴胺(DA)、高香草酸(HVA)、3,4-二羟基苯乙酸(DOPAC)和去甲肾上腺素(NE)的含量,用放射免疫法检测脑组织亮氨酸-脑啡肽(L-EK)和β-内啡呔(β-EP)的含量;采用小鼠抗焦虑实验和小鼠爬梯实验评价脑痛立停胶囊的抗焦虑作用。
结果:
1.无论现代处方还是古代方剂,祛风散寒止痛药都是使用频率最高的药物。用于治疗偏头痛的现代处方和古代方剂中,川芎、白芷和细辛都是最常用的药物。从功效方面看,用于治疗偏头痛的现代处方中,多以活血化瘀,通络止痛,祛风止痛,平肝息风为主。
2.电刺激SSS区硬脑膜偏头痛模型大鼠脑干和下丘脑c-fos、c-jun阳性细胞数及阳性细胞面积明显高于正常对照组和假手术组,经脑痛立停胶囊灌胃治疗后,模型大鼠脑干和下丘脑c-fos、c-jun阳性细胞数及阳性细胞面积明显下降,其中大剂量组作用明显,其作用强度与萘普生相当。
3.电刺激SSS区硬脑膜偏头痛模型大鼠血浆CGRP、ET含量明显升高,脑痛立停胶囊可明显降低模型动物血浆CGRP水平。但对升高的血浆ET无明显影响。4. ICR小鼠对利血平的反应较为敏感,与文献报道的KM小鼠在利血平使用剂量方面有很大差异。本实验皮下注射利血平0.25mg/kg方法复制的偏头痛小鼠模型血浆和脑组织5-HT、NE、DA含量较正常小鼠明显减少,说明模型复制成功。
5.利血平化低5-HT偏头痛模型小鼠体重下降、体温降低、凝血时间缩短,血浆及脑组织5-HT、DA的含量明显降低,脑组织NE、和HVA含量也明显降低。脑痛立停胶囊可以改善利血平化低5-HT偏头痛模型小鼠的一般状况,提高模型小鼠的体重、体温并延长模型小鼠的凝血时间;可提高模型小鼠血浆和脑组织5-HT、DA及脑组织NE的含量,同时降低模型小鼠血浆DOPAC含量,作用强度与复方羊角胶囊相似;脑痛立停胶囊对模型小鼠血浆5-HIAA和HVA含量无显著影响。
6.利血平化低5-HT偏头痛模型小鼠血浆和脑组织5-HT、DA和脑组织NE含量下降的同时,脑组织L-EK含量也明显低于正常对照组和假手术组。脑痛立停胶囊在提高利血平化低5-HT偏头痛模型小鼠血浆和脑组织5-HT、DA含量和脑组织NE的同时,能够明显提高模型小鼠脑组织L-EK和β-EP的含量。7.脑痛立停胶囊可以增加小鼠抗焦虑实验跨箱次数,缩短黑箱停留时间;同时可增加爬梯实验的爬梯级数,减少小鼠直立次数。
结论:
1.脑痛立停胶囊可能通过以下机制发挥短期缓解疼痛作用:通过某些作用机制抑制脑干、下丘脑c-fos与c-jun基因的表达,减弱或阻断了疼痛刺激在中枢神经的传导;降低血浆CGRP的含量,从而发挥调节血管舒缩障碍及抗炎作用;提高血浆及脑组织中5-HT、DA和脑组织NE的含量而起到调整脑部血管舒缩功能作用;调节血小板功能,减轻血液高凝状态,改善脑部血液循环。
2.脑痛立停胶囊可能通过以下机制发挥预防偏头痛发作的长期治疗作用:提高脑组织5-HT、NE、DA、L-EK和β-EP的含量,加强偏头痛患者中枢痛调制系统的功能;改善偏头痛患者的焦虑症状。
Objective
To discuss the action mechanism of Naotongliting capsule as a new medicine used to treat patients with migraine both on long term protective function and immediate effect of alleviating pain.
Methods
Both ancient and modern traditional Chinese medicine (TCM) prescriptions used to cure migraine are collected for finding the general rule of herb selection among those prescriptions by statistical analyzing;After making rat model with migraine by electrostimulating superior sagittal sinus(SSS),the model rats are treated whit Naotongliting capsule meanwhile taking Naproxen Sodium as positive control .The protein products of c-fos and c-jun gene in brine stem and hypothalamus of rats are tested with immunohistochemical method SABC.The content of calaitonin gene related peptide(CGRP) and endothelin(ET) in blood plasma of rats is measured by means of radioimmunity test; Mice model with migraine caused by exhausting 5-hydroxytryptamine(5-HT) with Reserpine is built up ,then the model mice are treated whit Naotongliting capsule.Fufangyangjiao capsule is used as control medicine.The content of 5-HT, 5-hydroxyindole acetic acid(5-HIAA),dopamine(DA),homovanillic acid(HVA),dihydroxyphenylacetic acid(DOPAC)and norepinephrine(NE)in brain and blood plasma of mice are tested by electrochemical meathod.The level of leucine enkephalin(L-EK) andβ- endorphin(β-EP) in brain of mice is tested with radioimmunity method;Staircase test and anti-anxiety test are used to estimate the dissipate anxiety effect of Naotongliting capsule
Results
1. The herbal medicine which have functions of dispelling wind-evil, dispersing cold and alleviating pain are most widely used not only in ancient TCM prescriptions, but also modern TCM prescriptions. Among the TCM prescriptions treating migraine.Chuanxiong, Baizhi and Xixin are used most frequently, and most of those prescriptions have efficacy virtue of stimulating circulation to end stasis, expelling the obstruction from collaterals and calming the liver to stop the wind.
2. Model rats with migraine caused by electrostimulating SSS have more cells which express protein products of c-fos and c-jun gene in brine stem and hypothalamus ,and the area of c-fos and c-jun positive cells is higher than normal control group and false operation group. Naotongliting capsule can decrease the mumber and the area of c-fos and c-jun positive cells of model rats.Big dose therapy group has better effect that is similar to Naproxen Sodium.
3. The level of CGRP and ET in blood plasma of model rats with migraine caused by electrostimulating SSS is upregulated. Naotongliting capsule can downregulate the level of CGRP in blood plasma of model rats,but it has no obvious effect on upregulated ET level .
4. According to the results of this research, ICR mouse is sensitive to Reserpine.The dose of Reserpine ICR mouse can endureis quite different to this of KM mouse that is reported in articles. In this experiment ,the dose of Reserpine giving to ICR mouse is 0.25mg/kg when causing mice model with migraine successfuly.The content of 5-HT,DA and NE in blood plasma and brain stem and hypothalamus is decreased obviously.
5. Model mice with migraine caused by exhausting 5-HT with Reserpine have low temperature and low body weight and short blood clotting time.The content of 5-HT,DA in blood plasma ,brain stem and hypothalamus of model Mice is decreased ,and NE,HVA in brine stem and hypothalamus is decreased too. Naotongliting capsule can better the general condition of model mice, increase temperature and body weight, prolong blood clotting time of model mice;Meanwhile the content of 5-HT,DA in blood plasma ,brain stem and hypothalamus of model mice are increased,and the content of DOPAC in plasma is downregulated. The effect is similar to Fufangyangjiao capsule.But Naotongliting capsule has no obvious effect on 5-HIAA and DA in plasma.
6. Model mice with migraine caused by exhausting 5-HT with Reserpine not only have lower content of 5-HT and DA in plasma, brain stem and hypothalamus, lower content of NE in brine stem and hypothalamus,but also have lower level of L-EK andβ-EP in brain than normal control and false injection groups. Naotongliting capsule can also upregulate the level of L-EK andβ-EP in brine, when upregulate the content of 5-HT and DA in plasma and brain stem and hypothalamus, upregulate the content of NE in brain of model mice.There is an obvious relation between effect and dose.
7. The Naotongliting capsule can increases frequency with which the mice stride over the light-dark chambers,and abbreviate the time of mice staying in the dark chamber;Meanwhile It can increases the number of staircases that the mice clamber,and decreases the frequency with which the mice arises.
Conclusions
1. Naotongliting capsule could give its immediate effect of alleviating pain by following mechanism:restraining the expression of c-fos and c-jun gene by some kind of way, weakening or stopping the conduction of pain stimulation in the center nerves system;Downregulating the level of CGRP in plasma to regulate the function of blood vessel and improv anti-inflammatory effect;Upregulating the level of 5-HT and DA in plasma , brain stem and hypothalamus, increasing the content of NE in brine stem and hypothalamus,to regulate function disorder of blood vessel in brain;Regulating function of thrombocyte, abating the blood clotting condition to improving the blood circulation in brain.
2. Naotongliting capsule could give its long term protective function by following mechanism: Upregulating the level of 5-HT, NE, DA, L-EK andβ-EP in brain,to improve the function of pain modulation of the center nerves system; Calming the anxiety symptoms of patients with migraine.
引文
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