CN115814041B - A Chinese medicinal composition for treating headache, and its preparation method - Google Patents
A Chinese medicinal composition for treating headache, and its preparation method Download PDFInfo
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- CN115814041B CN115814041B CN202211676272.4A CN202211676272A CN115814041B CN 115814041 B CN115814041 B CN 115814041B CN 202211676272 A CN202211676272 A CN 202211676272A CN 115814041 B CN115814041 B CN 115814041B
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses a traditional Chinese medicine composition for treating headache and a preparation method thereof, belonging to the field of traditional Chinese medicines. The traditional Chinese medicine composition consists of 3 parts: 1) Rhizoma Ligustici Chuanxiong, rhizoma Cyperi preparata, and Curcuma rhizome volatile oil clathrate; 2) Water extract composition of Olibanum, myrrha, radix Clematidis, radix Cyathulae, radix Angelicae sinensis, and radix astragali Preparata; 3) Safflower and red sage alcohol extract composition. Wherein, the mass ratio of the szechuan lovage rhizome, the prepared nutgrass galingale rhizome, the turmeric, the prepared frankincense, the prepared myrrh, the clematis root, the medicinal cyathula root, the Chinese angelica, the roasted astragalus, the safflower and the red sage root is as follows: 12-17:10-14:8-12:5-8:5-8:12-17:12-17:8-12:12-17:8-12:18-22. The traditional Chinese medicine composition provided by the invention can obviously improve the headache severity and the attack frequency of headache patients, improves the life quality of the patients, and is safe and free from toxic and side effects.
Description
Technical Field
The invention belongs to the field of traditional Chinese medicines, and in particular relates to a traditional Chinese medicine composition for treating headache and a preparation method thereof.
Background
The disclosure of this background section is only intended to increase the understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art already known to those of ordinary skill in the art.
Headache is a clinically common symptom and will generally be localized to the upper part of the skull, including the arch of the eyebrow, the upper edge of the auricle and the pain above the occipital protuberance line, collectively referred to as headache. The causes of headache are numerous and can be broadly divided into primary and secondary. The former cannot be attributed to a certain exact etiology, and can also be called idiopathic headache, such as migraine and tension headache; the latter etiology may involve various intracranial lesions such as cerebrovascular diseases, intracranial infections, craniocerebral trauma, systemic diseases such as fever, internal environmental disorders, abuse of psychoactive drugs, etc. The headache referred to by the invention mainly refers to primary headache such as migraine, tension type headache, angioneurotic headache and the like.
At present, the cause of migraine is not clear, western medicines are mainly used for treatment, the symptoms and root causes are not cured although the medicines relieve the symptoms to a certain extent, long-term medicine taking is usually required, once the medicine is stopped, headache symptoms can be generated, if the medicine is improperly taken, the illness state can be invalid or even aggravated, the toxic and side effects of the western medicines are large, especially, the influence on liver and kidney functions is the greatest, the illness state is repeated, and the side effects of the medicines bring great pain to patients.
Disclosure of Invention
In order to solve the defects of the prior art, the invention aims to provide the traditional Chinese medicine composition for treating headache and the preparation method thereof.
In order to achieve the above purpose, the technical scheme of the invention is as follows:
in a first aspect of the present invention, there is provided a Chinese medicinal composition for treating headache, the Chinese medicinal composition comprising the following components: 1) Rhizoma Ligustici Chuanxiong, rhizoma Cyperi preparata, and Curcuma rhizome volatile oil clathrate; 2) Water extract composition of Olibanum, myrrha, radix Clematidis, radix Cyathulae, radix Angelicae sinensis, and radix astragali Preparata; 3) Safflower and red sage alcohol extract composition.
Ligusticum wallichii has the effects of promoting blood circulation, activating qi-flowing, dispelling wind and relieving pain. Is a blood qi-flowing medicine, is spicy and fragrant to move qi, activate blood and remove stasis to stop bleeding, and ascend head and eyes to dispel wind, and is suitable for various pains caused by qi stagnation and blood stasis; it is used for treating headache, rheumatalgia, wind-cold headache, migraine, vascular neuralgia, etc. Is also one of the single traditional Chinese medicines with highest frequency of clinical treatment of headache, vascular headache and migraine. The main component contains volatile oil (mainly phthalides), phenolic acids and alkaloids, and other chemical components such as ligustrazine, sodium ferulate, ligustilide, phenylamide I, butylphthalide, etc. Wherein sodium ferulate and ligustrazine are main components of rhizoma Ligustici Chuanxiong. Modern pharmacological researches have proved that the volatile oil of Ligusticum wallichii has the effects of improving microcirculation, lowering blood pressure, increasing cerebral blood flow, relieving pain, regulating cardiovascular function, resisting blood coagulation, etc. Ligustrazine is used for treating migraine and cerebrovascular diseases clinically, can well reduce peripheral resistance of blood vessels, resists vasomotor, and has the effect of inhibiting platelet accumulation. Ligustrazine also has antiinflammatory, antioxidant and neuroprotective effects. Ligustilide has remarkable anti-migraine activity.
Rhizoma Cyperi has effects of dispersing stagnated liver qi for relieving qi stagnation, regulating qi-flowing for relieving rigidity of middle warmer, regulating menstruation and relieving pain; the main components are volatile oil, flavonoids, terpenes, phenols, alkaloids and other compounds; proved by researches, the nutgrass galingale rhizome has the pharmacological effects of resisting bacteria, easing pain, resisting cancer, reducing blood sugar, resisting blood acid and the like. The main component alpha-cyperus ketone of the cyperus volatile oil has stronger antipyretic and analgesic effects. Turmeric has effects of breaking blood, promoting qi circulation, dredging channels and relieving pain; the main components comprise curcumin, turmeric volatile oil and the like, wherein the volatile oil accounts for 4.0% -6.0%; the volatile oil has the effects of inhibiting tumor, enhancing immunity, and relieving pain. The volatile oil components of the ligusticum wallichii, the rhizoma cyperi and the turmeric have good pain relieving effect.
The red sage root has the effects of promoting blood circulation, removing blood stasis, dredging channels, relieving pain, clearing heart fire, relieving restlessness, cooling blood and eliminating carbuncles; the effective components of Saviae Miltiorrhizae radix include liposoluble components such as tanshinone IIA, cryptotanshinone, tanshinone I, etc., and water soluble components such as salvianolic acid B, rosmarinic acid, tanshinol, protocatechuic aldehyde, etc.; safflower has the effects of activating blood, dredging channels, removing stasis and relieving pain; the main components comprise flavone, alkaloid, polyacetylene, lignan, sesquiterpene, polysaccharide, etc. The red sage root and the safflower are extracted by alcohol, which is beneficial to the extraction of the active ingredients.
The active ingredients of the prepared frankincense and the prepared myrrh are resin ingredients; the effective components of the clematis root are saponin components and the like, and the medicinal materials of the medicinal cyathula root, the Chinese angelica and the roasted astragalus root are tonic, so that the medicinal materials adopt a water decoction mode, and the medicinal effect is better exerted.
Preferably, the Chinese medicinal composition comprises the following components in percentage by mass: 12-17:10-14:8-12:5-8:5-8:12-17:12-17:8-12:12-17:8-12:18-22.
Preferably, the traditional Chinese medicine composition is a tablet, a capsule, a granule or an enteric preparation.
In a second aspect, the present invention provides a preparation method of the above-mentioned Chinese medicinal composition for treating headache, which comprises the following steps:
s1, extracting volatile oil from rhizoma ligustici wallichii, prepared nutgrass galingale rhizome and turmeric, and preparing a volatile oil inclusion compound;
s2, extracting prepared frankincense, prepared myrrh, radix clematidis, medicinal cyathula root, chinese angelica and roasted astragalus root with water to obtain a water extraction composition;
s3, extracting the prepared safflower and the red sage root with ethanol to obtain an ethanol extract composition;
s4, mixing the volatile oil inclusion compound, the water extraction composition and the alcohol extraction composition.
Preferably, the extracting steps of the volatile oil are as follows: extracting rhizoma Ligustici Chuanxiong, rhizoma Cyperi preparata, and Curcuma rhizome with steam for 2-6 hr, and collecting volatile oil and steam water decoction.
Preferably, the specific steps for preparing the volatile oil inclusion compound are as follows: mixing the steam water decoction and beta-cyclodextrin in a container, stirring, adding into colloid mill, adding volatile oil under grinding, clathrating for 30-45 min, collecting condensate water, and controlling clathrating temperature between 30-40deg.C; preferably, the mass ratio of the beta cyclodextrin to the volatile oil is 6:1.
Preferably, after volatile oil extraction is carried out on the ligusticum wallichii, the prepared rhizoma cyperi and the turmeric, water extraction is carried out on dregs after the volatile oil extraction is carried out.
Further preferably, the specific steps of water extraction are as follows: extracting the residue with water for 2 times, 8-10 times of the water for 1-2 hr, and 6-8 times of the water for 0.5-1 hr, mixing filtrates, concentrating to 0.5 times of the amount of the medicinal materials, and mixing with the water extract composition obtained in step S2.
Preferably, in step S2, the specific water extraction step is as follows: extracting Olibanum, myrrha, radix Clematidis, radix Cyathulae, radix Angelicae sinensis, and radix astragali Preparata with water for 2 times, reflux extracting with 6-8 times of water under heating for 1-2 hr, extracting with 4-6 times of water for 1-1.5 hr, mixing filtrates, concentrating, removing impurities, and concentrating under reduced pressure to 0.5 times of medicinal materials to obtain water extract composition.
Preferably, in step S3, the specific ethanol extraction step is as follows: extracting Carthami flos and Saviae Miltiorrhizae radix with 70% ethanol for 2 times, extracting with 6-8 times of 70% ethanol for 1-2 hr, extracting with 4-6 times of 70% ethanol for 0.5-1 hr, mixing filtrates, recovering ethanol, and concentrating to 0.5 times of the amount of the medicinal materials to obtain ethanol extract composition.
The beneficial effects of the invention are as follows:
the traditional Chinese medicine composition can correct the level abnormality of monoamine neurotransmitters in migraine attacks by down-regulating the levels of NO, CGRP, 5-HT and DA in serum, so that the release and metabolic mechanism of the monoamine neurotransmitters of organisms are restored, the neurotransmitters are combined with corresponding receptors or target sites, the brain dysfunction is improved, the vasomotor dysfunction is improved, and the migraine induced by nitroglycerin is effectively prevented.
The traditional Chinese medicine composition provided by the invention can obviously improve the headache severity and the attack frequency of headache patients, improves the life quality of the patients, and is safe and free from toxic and side effects.
Detailed Description
In order to enable those skilled in the art to more clearly understand the technical scheme of the present invention, the technical scheme of the present invention will be described in detail with reference to specific embodiments.
Example 1
The traditional Chinese medicine composition for treating headache consists of the following raw materials in parts by weight: 12-17 parts of ligusticum wallichii, 10-14 parts of prepared rhizoma cyperi, 8-12 parts of turmeric, 5-8 parts of prepared frankincense, 5-8 parts of prepared myrrh, 12-17 parts of radix clematidis, 12-17 parts of medicinal cyathula root, 8-12 parts of Chinese angelica, 12-17 parts of radix astragali preparata, 8-12 parts of safflower and 18-22 parts of radix salviae miltiorrhizae.
The preparation method comprises the following steps:
the traditional Chinese medicine composition consists of 3 parts: 1) Rhizoma Ligustici Chuanxiong, rhizoma Cyperi preparata, and Curcuma rhizome volatile oil clathrate; 2) Water extract composition of Olibanum, myrrha, radix Clematidis, radix Cyathulae, radix Angelicae sinensis, and radix astragali Preparata; 3) Mixing Carthami flos and Saviae Miltiorrhizae radix ethanol extract composition, and mixing the 3 parts. The optimal technological parameters are screened through the detection of the content of the main component.
1. Preparation method of volatile oil clathrate of rhizoma Ligustici Chuanxiong, rhizoma Cyperi preparata and Curcuma rhizome
1.1 extraction Process screening
1) Extracting volatile oil: extracting rhizoma Ligustici Chuanxiong, rhizoma Cyperi preparata, and Curcuma rhizome with steam for 2-6 hr, and collecting volatile oil and small amount of steam water decoction.
2) Water extraction: extracting the residue with water for 2 times, 8-10 times of water for 1-2 hr, and 6-8 times of water for 0.5-1 hr, mixing filtrates, concentrating under reduced pressure to 0.5 times of medicinal materials, and mixing with the water extract composition.
3) Preparing volatile oil inclusion compound: mixing the steam water decoction and beta-cyclodextrin (beta cyclodextrin: volatile oil mass ratio of 6:1), stirring, adding into colloid mill, adding volatile oil under grinding, clathrating for 30-45 min, adding condensed water, and clathrating at 30-40deg.C to obtain clathrate.
2. Route for the preparation of water-extractable compositions
2.1 extraction Process screening
2.1.1 optimal process parameters:
reflux extracting Olibanum, myrrha, radix Clematidis, radix Cyathulae, radix Angelicae sinensis, and radix astragali Preparata with 8 times of water under heating for 2 hr, extracting with 6 times of water for 1 hr, filtering, mixing filtrates, concentrating, centrifuging with high-speed centrifuge to remove macromolecular impurities, concentrating under reduced pressure to 0.5 times of medicinal materials, and mixing with the above rhizoma Ligustici Chuanxiong, rhizoma Cyperi preparata, and Curcuma rhizome water extract to obtain water extract composition.
2.1.2 Process optimization experiments
Paste yield = dry paste amount/crude drug amount 100 ×
Transfer rate = dry extract amount x dry extract content/index ingredient-containing medicinal material amount x 100 of medicinal material content
2.1.2.1 Water addition multiple investigation
Respectively taking 60g of prepared frankincense, 60g of prepared myrrh, 150g of radix clematidis, 150g of medicinal cyathula root, 150g of Chinese angelica and 150g of roasted astragalus, taking 3 parts by weight, and respectively performing a process experiment according to the following steps.
(1) Decocting with 6 times of water for 2 hours, and decocting with 6 times of water for 1 hour;
(2) 8 times of water is decocted for 2 hours, and 6 times of water is decocted for 1 hour;
(3) 10 times of water is decocted for 2 hours, and 8 times of water is decocted for 1 hour.
Extracting according to the above process, filtering, mixing filtrates, concentrating under reduced pressure until the medicinal materials are heavy, and drying to obtain a sample, detecting according to index components, with the following results:
TABLE 1
Conclusion: the paste yield and the transfer rate of each component of the process (1) are low, the paste yield and the transfer rate of the processes (2) and (3) are high, and the process (2) is selected in combination with the production cost, namely 8 times of mass water is decocted first and 6 times of mass water is decocted second.
2.1.2.2 extraction time investigation
Respectively taking 60g of prepared frankincense, 60g of prepared myrrh, 150g of radix clematidis, 150g of medicinal cyathula root, 150g of Chinese angelica and 150g of roasted astragalus, taking 3 parts by weight, and respectively performing a process experiment according to the following steps.
(4) 8 times of water is decocted for 1.5 hours, and 6 times of water is decocted for 0.5 hours;
(5) 8 times of water is decocted for 2 hours, and 6 times of water is decocted for 1 hour;
(6) 8 times of water is decocted for 2.5 hours, and 6 times of water is decocted for 1.5 hours.
Extracting according to the above process, filtering, mixing filtrates, concentrating under reduced pressure until the medicinal materials are heavy, drying to obtain sample, detecting according to index components, and calculating the ointment rate and transfer rate of each index component, with the following results:
TABLE 2
Conclusion: the paste yield and the transfer rate of each component of the process (4) are low, the paste yield and the transfer rate of the processes (5) and (6) are high, and the process (5) is selected in combination with the production cost, namely one decoction for 2 hours and two decoction for 1 hour.
In summary, the water extract is best prepared by the following steps: 8 times of water is decocted for 2 hours, and 6 times of water is decocted for 1 hour.
2.2 index component detection
The optimal extraction process parameters are screened by measuring the content of oleanolic acid, cuparasterone, ferulic acid and astragaloside in the main active ingredients of radix clematidis, radix cyathulae, angelica and radix astragali preparata.
2.2.1 determination of oleanolic acid, calaranthrone and Ferulic acid content
Chromatographic conditions: the column was Phenomenext Luna C column (250 mm. Times.4.6 mm,5 μm); the mobile phase is acetonitrile (A) -0.085% phosphoric acid solution (B), and gradient elution is carried out: 0 to 20min,10 to 30 percent of acetonitrile; 20-45 min, 30-57% acetonitrile; 45-60 min, 57-85% acetonitrile, and detection wavelength of 240nm; the flow rate is 1.0mL min < -1 >; the column temperature is 30 ℃; the sample loading was 10. Mu.L.
Preparation of a control solution: 3 reference substances of oleanolic acid, cuparanthrone and ferulic acid are precisely weighed, and mixed reference substance solutions containing 30, 15 and 20 mug of oleanolic acid, cuparanthrone and ferulic acid are prepared by using 80% methanol in each 1 ml.
Preparation of test solution: taking about 1.0g of extract, precisely weighing, precisely adding 30ml of 80% methanol solution, weighing, refluxing for 1h, taking out, cooling, weighing again, supplementing the lost weight with 80% methanol, shaking, and filtering.
And (3) measuring: respectively precisely sucking 10 μl of the reference solution and the sample solution, and injecting into a liquid chromatograph for measurement.
2.2.2 determination of Astragaloside IV content
Chromatographic conditions and system suitability test: octadecylsilane chemically bonded silica is used as a filler; acetonitrile-water (32:68) as mobile phase; the evaporative light scattering detector detects. The theoretical plate number is not less than 4000 calculated according to astragaloside IV peak.
Preparation of a control solution: taking appropriate amount of astragaloside IV reference substance, precisely weighing, and adding 80% methanol to obtain solution containing 50 μg per 1 ml.
Preparation of test solution: taking about 1g of extract, precisely weighing, placing into a conical flask with a plug, precisely adding 80% methanol solution containing 4% of concentrated ammonia test solution (taking 4ml of concentrated ammonia test solution, adding 80% methanol to 100ml, shaking uniformly), sealing, weighing, heating and refluxing for 1 hour, cooling, weighing again, supplementing the lost weight with 80% methanol solution containing 4% of concentrated ammonia test solution, shaking uniformly, filtering, precisely measuring 15ml of subsequent filtrate, evaporating, dissolving residues with 80% methanol, transferring to a 5ml measuring flask, adding 80% methanol to scale, shaking uniformly, filtering, and taking the subsequent filtrate.
Assay: 2 μl (or 5 μl) of the reference solution and 10 μl of the sample solution are respectively precisely sucked, 10-20 μl of the sample solution is injected into a liquid chromatograph for measurement, and the external standard two-point method logarithmic equation is used for calculation, thus obtaining the final product.
3. Route for the preparation of alcohol extraction compositions
3.1 extraction Process screening
3.1.1 optimal process parameters:
extracting Carthami flos and Saviae Miltiorrhizae radix with 70% ethanol for 2 times, 7 times of 70% ethanol for 1.5 hr, and 5 times of 70% ethanol for 1 hr, mixing filtrates, recovering ethanol, and concentrating to 0.5 times of the amount of the medicinal materials to obtain ethanol extract.
3.1.2 ethanol concentration investigation
100g of safflower and 200g of red sage root are respectively taken, 3 parts of safflower and 200g of red sage root are respectively taken by the same weight, and the process experiments are respectively carried out according to the following procedures.
(7) Reflux-extracting with 80% ethanol 7 times for 1.5 hr, reflux-extracting with 80% ethanol 6 times for 1 hr;
(8) Reflux-extracting with 70% ethanol 7 times for 1.5h, reflux-extracting with 70% ethanol 6 times for 1h;
(9) Reflux-extracting with 7 times of 60% ethanol for 1.5h, and reflux-extracting with 6 times of 60% ethanol for 1h.
Extracting according to the above process, filtering, mixing filtrates, concentrating under reduced pressure, recovering ethanol until no ethanol smell is present, drying to obtain sample, and detecting according to index components to obtain the following results:
TABLE 3 Table 3
Conclusion: the paste yield of the 3 processes is basically no difference, the index component transfer rates of the processes (7) and (8) are relatively high, and the process (8) is selected in combination with the production cost, namely 70% ethanol reflux extraction is selected.
2.2.1.2 Investigation of the amount of solvent extracted
100g of safflower and 200g of red sage root are respectively taken, 3 parts of safflower and 200g of red sage root are respectively taken by the same weight, and the process experiments are respectively carried out according to the following procedures.
(10) Reflux-extracting with 70% ethanol 9 times for 1.5 hr, and reflux-extracting with 80% ethanol 7 times for 1 hr;
(11) Reflux-extracting with 70% ethanol 7 times for 1.5h, reflux-extracting with 70% ethanol 6 times for 1h;
(12) Reflux-extracting with 5 times of 70% ethanol for 1.5h, and reflux-extracting with 4 times of 70% ethanol for 1h.
Extracting according to the above process, filtering, mixing filtrates, concentrating under reduced pressure, recovering ethanol until no ethanol smell is present, drying to obtain sample, and detecting according to index components to obtain the following results:
TABLE 4 Table 4
Conclusion: the process (10) has lower paste yield and transfer rate of each component, the processes (11) and (12) have higher paste yield and transfer rate, and the process (11) is selected in combination with the production cost, namely 7 times of 70% ethanol is selected for the first time of reflux, and 6 times of 70% ethanol is selected for the second time of reflux.
2.2.1.3 Investigation of extraction time
100g of safflower and 200g of red sage root are respectively taken, 3 parts of safflower and 200g of red sage root are respectively taken by the same weight, and the process experiments are respectively carried out according to the following procedures.
(13) Reflux-extracting with 70% ethanol 7 times for 1h, reflux-extracting with 70% ethanol 6 times for 0.5h;
(14) Reflux-extracting with 70% ethanol 7 times for 1.5h, reflux-extracting with 70% ethanol 6 times for 1h;
(15) Reflux-extracting with 7 times of 70% ethanol for 2h, and reflux-extracting with 7 times of 70% ethanol for 1.5h.
Extracting according to the above process, filtering, mixing filtrates, concentrating under reduced pressure, recovering ethanol until no ethanol smell is present, drying to obtain sample, and detecting according to index components to obtain the following results:
TABLE 5
Conclusion: the paste yield and the transfer rate of each component of the process (13) are low, the paste yield and the transfer rate of the processes (14) and (15) are high, and the process (14) is selected in combination with the production cost, namely 7 times of 70% ethanol is subjected to reflux extraction for 1.5h, and 6 times of 70% ethanol is subjected to reflux extraction for 1h.
In summary, the alcohol extract is best prepared by the following steps: reflux-extracting with 70% ethanol 7 times for 1.5 hr, and reflux-extracting with 70% ethanol 6 times for 1 hr.
3.2 index component detection
The content of hydroxysafflor yellow A, salvianolic acid B, cryptotanshinone and tanshinone IIA in the main active components of safflower and red sage root is measured, and the optimal extraction technological parameters are screened.
Chromatographic conditions: the column was Phenomenext Luna C column (250 mm. Times.4.6 mm,5 μm); the mobile phase is acetonitrile (A) -0.1% phosphoric acid solution (B), and gradient elution is carried out: 0-10 min, 10-10% acetonitrile; 10-40 min, 10-22% acetonitrile; 40-50 min, 22-22% acetonitrile; 50-65 min, 22-50% acetonitrile; 65-100 min, 50-85% acetonitrile; the detection wavelength is 205nm; the flow rate is 1.0mL min < -1 >; the column temperature is 30 ℃; the sample loading was 10. Mu.L.
Preparation of a control solution: the method comprises precisely weighing 4 reference substances of hydroxysafflor yellow A, salvianolic acid B, cryptotanshinone and tanshinone IIA, and preparing mixed reference substance solutions containing hydroxysafflor yellow A, salvianolic acid B, cryptotanshinone and tanshinone IIA 50, 15, 20 and 10 μg per 1ml with 80% methanol.
Preparation of test solution: taking about 1.0g of extract, precisely weighing, precisely adding 30ml of 80% methanol solution, weighing, refluxing for 1h, taking out, cooling, weighing again, supplementing the lost weight with 80% methanol, shaking, and filtering.
And (3) measuring: respectively precisely sucking 10 μl of the reference solution and the sample solution, and injecting into a liquid chromatograph for measurement.
Application example 1
Study of the efficacy of the Chinese medicinal composition for treating headache obtained in example 1
The method comprises the steps of establishing a chronic migraine animal model by injecting nitroglycerin into the abdominal cavity of a mouse at a low dose for many times, recording the frequency of the front limb of the mouse, detecting biochemical indexes of Nitric Oxide (NO), dopamine (DA), calcitonin Gene Related Peptide (CGRP) and 5-hydroxytryptamine (5-HT) of the mouse, and carrying out pharmacodynamics experimental study on the composition for treating migraine.
The nitroglycerin can enhance the excitability of ash neurons around the tail of the trigeminal nerve and the midbrain aqueduct, improve the afferent reaction of the spinal cord dorsal horn to the nociceptive information, further lead the afferent branch of the trigeminal nerve which innervates the meninges to generate delayed and long-acting pain sensitization, and reduce the threshold value of pain sensation. The nitroglycerin is one of the donor substances of NO, can stimulate the organism to promote vascular smooth muscle to release NO to cause intracranial and extracranial vascular dilation so as to produce headache, and simultaneously, the NO activates guanylate cyclase through dispersion action, so that vascular smooth muscle is promoted to relax, and the intracranial and extracranial vascular dilation is caused to cause headache. In addition, NO can mediate the release of various neuropeptides such as calcitonin gene-related peptide, trigger neurogenic inflammation, and participate in migraine attacks. Administration can be by subcutaneous injection, intravenous injection, and intraperitoneal injection, with intraperitoneal injection being the most common. The acute and chronic migraine of the people can be simulated by adjusting the quantity of single injection. NO, DA, CGRP and 5-HT are important biochemical indicators for the detection of migraine. NO has neurotoxic effects that trigger trigeminal-vascular reflex, inducing experimental migraine. 5-HT is an important neurotransmitter in migraine attacks, one of the total essential neuroactive substances in the central endogenous analgesic system, and increased release during migraine attacks can cause endorphin release in the pituitary to participate in pain regulation, associated with central nervous system pain transmission and endogenous analgesic mechanisms; DA is a neurotransmitter next to 5-HT in migraine attacks, and is also released to increase during migraine attacks, closely related to migraine attacks; CGRP is the predominant neuropeptide associated with the trigeminal system and has a strong vasodilating effect, causing headaches by vasodilating, and plays an important role in migraine attacks. The effect of the composition on treating headache is studied by detecting biochemical indexes such as NO, DA, CGRP, 5-HT and the like in mouse serum.
1. Animal and main reagent
SPF grade ICR mice were purchased from Liaoning long biotechnology Co., ltd in 60, male and female halves, body weight (18.+ -.2) g. Mice were fed habitually for 1 week, with free water and food prior to the experiment.
2. Experimental grouping
The experimental mice were randomly divided into 5 groups, control, model, high, medium, and low dose groups of 12 animals each. The high, medium and low dosage groups (high dosage 1.44g/kg, medium dosage 0.72g/kg, low dosage 0.36 g/kg) are respectively administered with the Chinese medicinal composition with different concentrations, and the physiological saline group (i.e. control group) and model group are administered with the same volume of physiological saline, and the stomach is continuously irrigated for 7d.
3. Model building
After 60min from the last administration, 5mg/kg of nitroglycerin injection was injected intraperitoneally into each experimental group 1 time every other day for 9 days in succession for 5 total injections. The control group was injected with the same volume of distilled water, resulting in an experimental migraine model. The administration was continued during the molding.
4. Observation of the number of times of mouse's scratching
After the last intraperitoneal injection of nitroglycerin for 30min, the number of times of scratching the forelimbs of each of the experimental group and the control group mice in 30min was recorded by a counter, and the results were recorded.
5. Biochemical index determination
After the end of the scratching test, blood was collected from the orbital veins of the mice in each of the administration group and the control group, and the blood was centrifuged at 3000 rpm for 10 minutes, the serum was separated, the NO content in the serum was measured by the nitrate reductase method, and the DA, CGRP, 5-HT content in the serum was measured by the ELISA method.
6. Experimental results
6.1 times of mouse scratching
Note that: * p < 0.05
6.2 Biochemical index determination
The level of each biochemical index in the dosing group was significantly reduced compared to the model group. There were significant differences in 5-HT levels in the lower dosing group compared to the model group; the levels of NO, 5-HT in the medium dose group were significantly different; there were significant differences in both high dose groups NO, DA, CGRP, 5-HT.
Note that: * p < 0.05
7. Conclusion(s)
According to the observation of the frequency of the mouse's head and the measurement of biochemical indexes (NO, DA, CGRP, 5-HT), the traditional Chinese medicine composition can correct the level abnormality of monoamine neurotransmitters in migraine attack by down-regulating the levels of NO, CGRP, 5-HT and DA in serum, so that the release and metabolic mechanism of the monoamine neurotransmitters in the organism can be restored, and the neurotransmitters can be combined with corresponding receptors or target sites to improve the brain dysfunction, improve vasomotor dysfunction and effectively prevent migraine induced by nitroglycerin.
Application example 2
Observation of case for treating headache
Clinical cases were collected and a comparison study was performed by VAS pain score on the treatment of headache with the composition formulation.
VAS pain scoring is an internationally recognized, general visual analog scoring method and patients self-evaluate clinical symptoms on a 10cm visual scale. 0 represents normal, without any pain, 10 represents extreme discomfort, very pain.
1. Case sources
During the period from month 1 2020 to month 12 2021, 82 patients diagnosed with wind-cold headache, wind-heat headache, migraine, angioneurotic headache were enrolled in total, ranging from 18-55 years of age, 27 men and 55 women. The granules prepared with the traditional Chinese medicine composition obtained in example 1 were used for 4 weeks of intervention.
2. Inclusion criteria
(1) Meets the diagnosis standard of wind-cold headache, wind-heat headache, migraine and angioneurotic headache in traditional Chinese medicine;
(2) Age 18-55 years;
(3) At least one headache per month for about 3 months;
(4) Voluntarily attending the study and signing informed consent.
3. Exclusion criteria
(1) Pregnant or lactating women;
(2) Combining serious brain, heart, lung, kidney, blood system diseases and malignant tumor except hypertension, diabetes and hyperlipidemia, and requiring drug therapy;
(3) Patients with infectious diseases, such as viral hepatitis, pulmonary tuberculosis, AIDS, etc.;
(4) Headache is secondary to craniocerebral and other organic lesions;
(5) Special headache, such as hemiplegia type headache, drug overdependence headache, etc.
4. VAS pain scoring
The patients were scored according to the VAS pain score scale and were classified into four categories.
Painless: the scoring value of the patient is less than or equal to 0 and less than or equal to 1, and the patient has no pain feeling.
Pain relieving: patient score of 1 is less than or equal to 4, patient is slightly painful, and general activities are not affected.
Pain in middle: the scoring value of the patient is less than or equal to 7 and is more than 4, and the patient has obvious pain and can exercise.
Heavy pain: patient score of 7 < 10, patient has severe pain with limited activity.
Pain improvement rate: (pre-treatment pain score-post-treatment pain score)/pre-treatment pain score x 100% analgesic efficacy criteria: the analgesic score is more than or equal to 60 percent; analgesic scores of 30% -60% are effective; the pain relieving score is less than or equal to 30 percent and is ineffective.
5. Comparison of effects before and after treatment
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Compared with the VAS score before treatment, 76 patients in 82 patients have reduced VAS score, and the analgesic improvement rate is more than 30 percent, namely the analgesic effect is effective and 66 patients with obvious effect, namely the patients account for 75 percent of the total patient number.
In conclusion, the pharmaceutical composition provided by the invention can effectively treat headache, remarkably improve the headache severity degree and the attack frequency of headache patients, improve the life quality of the patients and has a better treatment effect.
The above description is only of the preferred embodiments of the present invention and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (7)
1. A traditional Chinese medicine composition for treating headache, which is characterized by comprising the following components: 1) Rhizoma Ligustici Chuanxiong, rhizoma Cyperi preparata, and Curcuma rhizome volatile oil clathrate; 2) Water extract composition of Olibanum, myrrha, radix Clematidis, radix Cyathulae, radix Angelicae sinensis, and radix astragali Preparata; 3) Safflower and red sage alcohol extract composition;
the mass ratio of the szechuan lovage rhizome, the prepared nutgrass galingale rhizome, the turmeric, the prepared frankincense, the prepared myrrh, the clematis root, the medicinal cyathula root, the Chinese angelica, the roasted astragalus, the safflower and the red sage root is as follows: 12-17:10-14:8-12:5-8:5-8:12-17:12-17:8-12:12-17:8-12:18-22;
the preparation method of the traditional Chinese medicine composition for treating headache comprises the following steps:
s1, extracting volatile oil from rhizoma ligustici wallichii, prepared nutgrass galingale rhizome and turmeric, and preparing a volatile oil inclusion compound;
s2, extracting prepared frankincense, prepared myrrh, radix clematidis, medicinal cyathula root, chinese angelica and roasted astragalus root with water to obtain a water extraction composition;
s3, extracting the prepared safflower and the red sage root with ethanol to obtain an ethanol extract composition;
s4, mixing the volatile oil inclusion compound, the water extraction composition and the alcohol extraction composition;
the volatile oil extraction comprises the following specific steps: extracting rhizoma Ligustici Chuanxiong, rhizoma Cyperi preparata, and Curcuma rhizome with steam for 2-6 hr, and collecting volatile oil and steam water decoction;
the preparation method of the volatile oil inclusion compound comprises the following specific steps: mixing the steam water decoction and beta-cyclodextrin in a container, stirring, adding into colloid mill, adding volatile oil under grinding, clathrating for 30-45 min, collecting condensate water, and clathrating at 30-40deg.C.
2. The traditional Chinese medicine composition according to claim 1, wherein the traditional Chinese medicine composition is a tablet, a capsule, a granule or an enteric preparation.
3. The method of claim 1, wherein the mass ratio of beta cyclodextrin to volatile oil is 6:1.
4. The method according to claim 1, wherein the volatile oil is extracted from rhizoma Chuanxiong, rhizoma Cyperi preparata, and rhizoma Curcumae Longae, and the residue after the extraction is extracted with water.
5. The preparation method according to claim 4, wherein the specific steps of water extraction are as follows: extracting the residue with water for 2 times, 8-10 times of the water for 1-2 hr, and 6-8 times of the water for 0.5-1 hr, mixing filtrates, concentrating to 0.5 times of the amount of the medicinal materials, and mixing with the water extract composition obtained in step S2.
6. The preparation method according to claim 1, wherein in step S2, the specific steps of water extraction are as follows: extracting Olibanum, myrrha, radix Clematidis, radix Cyathulae, radix Angelicae sinensis, and radix astragali Preparata with water for 2 times, reflux extracting with 6-8 times of water under heating for 1-2 hr, extracting with 4-6 times of water for 1-1.5 hr, mixing filtrates, concentrating, removing impurities, and concentrating under reduced pressure to 0.5 times of medicinal materials to obtain water extract composition.
7. The method according to claim 1, wherein in step S3, the ethanol extraction comprises the following steps: extracting Carthami flos and Saviae Miltiorrhizae radix with 70% ethanol for 2 times, extracting with 6-8 times of 70% ethanol for 1-2 hr, extracting with 4-6 times of 70% ethanol for 0.5-1 hr, mixing filtrates, recovering ethanol, and concentrating to 0.5 times of the amount of the medicinal materials to obtain ethanol extract composition.
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