CN114390927A - New application of traditional Chinese medicine composition - Google Patents
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Abstract
A new application of a traditional Chinese medicine composition, in particular to an application of a traditional Chinese medicine composition in preparing a medicine for treating stroke. The Chinese medicinal composition is mainly prepared from leech, scorpion, centipede, angelica, figwort root, coix seed, phellodendron, honeysuckle, astragalus, liquorice and the like, and can also contain rhizoma atractylodis and white paeony root. Pharmacological test results show that the traditional Chinese medicine composition can effectively relieve secondary brain tissue injury of cerebral hemorrhage, and improve the behaviourology of rats with cerebral hemorrhage and stroke and the nerve function after cerebral hemorrhage; has effects of protecting cerebral tissue of ischemic apoplexy rat, and improving blood stasis symptom.
Description
The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to application of a traditional Chinese medicine composition in preparation of a medicine for treating stroke.
Apoplexy is a cerebral blood circulation disorder disease with sudden onset, also called cerebrovascular accident and cerebral apoplexy, and refers to acute cerebral blood circulation disorder caused by the stenosis, occlusion or rupture of internal cerebral arteries caused by various inducing factors. The disease has extremely high fatality rate and disability rate, is one of diseases with higher fatality rate in the world, and the mortality rate of the stroke also tends to rise with age, so that effective treatment measures are always lacked.
At present, the drugs for treating stroke in clinic are complicated, and examples include traditional Chinese medicines, vasodilators (such as dipyridamole, etc.), drugs for improving microcirculation and expanding blood volume (such as low molecular dextran, etc.), drugs for dissolving thrombus (such as urokinase, etc.), anticoagulant therapeutic drugs (such as heparin, etc.), drugs for preventing platelet aggregation (such as aspirin, etc.), and the like. The western medicine for treating the apoplexy sequelae mainly takes a nerve adjuvant drug as a main drug and also has the functions of reducing blood pressure, reducing blood fat and the like; while traditional Chinese medicine mainly takes medicines for promoting blood circulation and removing blood stasis, dredging collaterals and relieving pain and tonifying qi and blood, and is assisted by therapies such as acupuncture and moxibustion, massage and the like.
The Chinese invention patent CN1201787C discloses a medicine for treating thrombophlebitis, the product obtained by the patent technology has the functions of clearing away heat and toxic material, removing blood stasis and dredging collaterals, inducing diuresis and reducing edema, and has certain prevention and treatment effects on the treatment of thrombophlebitis and thromboangiitis obliterans. The preparation product prepared by the patent of the invention comprises the Mailuoshutong granules and the Mailuoshutong pills, and is widely used for clinically treating superficial thrombophlebitis caused by damp-heat stasis in veins and lower limb swelling, pain and dull red skin color caused by non-acute deep vein thrombosis or accompanied by cord-shaped substances, the clinical treatment efficiency is high, the pain of a patient can be effectively relieved, and the consistent favorable comment of the patient and doctors is obtained.
The invention relates to a new application of a traditional Chinese medicine composition developed on the basis of the Chinese patent of invention CN 1201787C.
The traditional Chinese medicine composition related in the patent can be prepared into medicines, in particular to Mailuoshutong granules/pills. The Mailushutong granule/pill series products are unique varieties of Lunan Kangpu pharmaceutical Co., Ltd, and zoology and clinical trials for new indications are also in progress.
The invention aims to provide a novel traditional Chinese medicine composition for treating stroke.
The traditional Chinese medicine composition is mainly prepared from leech, scorpion, centipede, Chinese angelica, figwort root, coix seed, amur corktree bark, honeysuckle flower, astragalus root, liquorice and the like; in addition, the raw materials can also comprise rhizoma atractylodis and white paeony root.
The raw materials are prepared from the following components:
leech 180 and 280 portions of scorpion 50-100 portions and centipede 10-40 portions
100 parts of angelica sinensis, 400 parts of radix scrophulariae, 300 parts of radix scrophulariae, 700 parts of semen coicis, 300 parts of semen coicis, 700 parts of semen coicis
100 parts of phellodendron amurense, 400 parts of honeysuckle, 300 parts of honeysuckle, 700 parts of astragalus, 300 parts of astragalus, 700 parts of astragalus
50-100 parts of liquorice, 100 parts of rhizoma atractylodis, 400 parts of white paeony root, 100 parts of white paeony root and 400 parts of white paeony root.
Preferably, the raw materials are prepared from the following components:
leech 180 and 280 portions of scorpion 50-100 portions and centipede 10-40 portions
300 parts of 200 parts of radix angelicae sinensis, 550 parts of 450 parts of radix scrophulariae, 550 parts of 450 parts of semen coicis
200 parts of phellodendron amurense, 450 parts of honeysuckle, 550 parts of astragalus mongholicus, 450 parts of astragalus membranaceus
50-100 parts of liquorice, 200 parts of rhizoma atractylodis, 200 parts of white paeony root and 280 parts of white paeony root.
In a further preferred scheme, the raw materials are prepared from the following components:
leech 250 parts, scorpion 83 parts, centipede 20 parts
280 parts of Chinese angelica, 530 parts of figwort root, 460 parts of coix seed
280 parts of golden cypress, 460 parts of honeysuckle flower, 530 parts of astragalus root
Licorice root 83 weight portions, atractylodes rhizome 220 weight portions, white peony root 220 weight portions.
In a further preferred embodiment, the raw materials are prepared from the following components:
leech 250 parts, scorpion 83 parts, centipede 20 parts
250 parts of Chinese angelica, 500 parts of figwort and 500 parts of coix seed
250 parts of phellodendron bark, 500 parts of honeysuckle flower and 500 parts of astragalus root
Licorice root 83 weight portions, atractylodes rhizome 250 weight portions, white peony root 250 weight portions.
Particularly, the Chinese angelica in the raw material composition is wine Chinese angelica, and the amur corktree bark is salt amur corktree bark.
According to dialectical thinking of traditional Chinese medicine, the main factors of the occurrence of the stroke are plain qi and blood deficiency, imbalance of yin and yang of the heart, the liver and the kidney, anxiety and anger, or alcohol drinking and eating, or invasion of exogenous pathogenic factors and the like of a patient, so that qi and blood circulation is blocked, and channels are blocked and are not nourished; or sudden collapse due to yin deficiency leading to sudden swelling of liver yang, transformation of yang into wind, adverse flow of qi due to blood, phlegm-fire, meridian cross, obstruction of orifices, hemiplegia, and apoplexy.
In the traditional Chinese medicine formula, leech, scorpion and centipede are taken as monarch drugs, and the traditional Chinese medicine formula is used for breaking blood and stimulating the menstrual flow, and removing blood stasis
The traditional Chinese medicine composition has the effects of dredging the channels and collaterals, removing blood stasis, smoothing qi and blood circulation, and treating the main symptoms of stroke, hemiplegia and the like caused by qi and blood circulation obstruction and channel blockage; angelica, astragalus, phellodendron and figwort are used as ministerial drugs, and the Chinese medicinal composition has the effects of activating blood and stimulating menstrual flow, cooling blood and lowering fire, and activating stagnancy and relieving arthralgia; coix seed, semen Coicis and honeysuckle flower, both of which have the actions of clearing heat and removing toxicity, inducing diuresis and excreting dampness, are used as adjuvant drugs; the liquorice is used as a guiding drug to harmonize the drug property and relieve the toxicity and the intensity of the drug. In addition to this, the present invention is,the rhizoma atractylodis and the white paeony root in the invention can dry dampness and invigorate spleen, and calm liver yang, and also have the function of adjuvant drugs.
The stroke of the invention is hemorrhagic stroke and/or ischemic stroke; preferably, the hemorrhagic stroke is cerebral hemorrhage and/or subarachnoid hemorrhage, and the ischemic stroke is cerebral infarction and/or cerebral thrombosis.
The apoplexy is mainly the apoplexy caused by diseases such as hypertension, atherosclerosis, cerebrovascular diseases, diabetes, hyperlipidemia and/or rheumatism, hypertension, coronary arteriosclerotic cardio-cerebral diseases and the like.
In addition, the traditional Chinese medicine composition can be prepared into clinically acceptable dosage forms;
preferably, the clinically acceptable dosage form is a tablet, a granule, a capsule or a pill;
further preferably, the dosage form is granules or pills;
more preferably, the granules are Mailuoshutong granules, and the pills are Mailuoshutong pills.
The preparation method of the Mailuoshutong granules comprises the following steps:
A. pulverizing part of Hirudo, Scolopendra, and Scorpio into fine powder;
B. distilling flos Lonicerae, rhizoma Atractylodis, radix scrophulariae, radix Angelicae sinensis, and radix Paeoniae alba with water, and extracting volatile oil; distilling the medicinal liquid and residue;
C. mixing the distillation residue obtained in the step B with radix astragali, cortex Phellodendri, Coicis semen, Glycyrrhrizae radix, the rest Hirudo, Scolopendra and Scorpio, and decocting with water to obtain decoction for use;
D. mixing the distilled liquid medicine obtained in the step B and the decoction obtained in the step C, concentrating, adding alcohol, standing, filtering, recovering ethanol, concentrating, drying, and crushing to obtain fine powder for later use;
E. and D, mixing the fine powder obtained in the step D and the fine powder obtained in the step A, adding conventional auxiliary materials, granulating, drying, and spraying the volatile oil obtained in the step B to obtain granules.
Specifically, the preparation method of the mailuoshutong granules comprises the following steps:
A. 1/2 pulverizing Hirudo, Scolopendra, and Scorpio into fine powder;
B. soaking flos Lonicerae, rhizoma Atractylodis, radix scrophulariae, radix Angelicae sinensis, and radix Paeoniae alba in water, distilling, and collecting volatile oil; distilling to obtain medicinal liquid and residue;
C. b, uniformly mixing the astragalus, the golden cypress, the coix seed, the liquorice, the residual 1/2 formula leech, the centipede and the scorpion with the distilled medicine residues obtained in the step B, adding water for decocting for 2 times, and mixing decoction for later use, wherein the decoction is 1.5 hours each time;
D. mixing the distilled liquid medicine obtained in the step B and the decoction obtained in the step C, filtering, concentrating the filtrate to obtain a clear paste with the relative density of 1.10-1.20 (80 ℃), adding ethanol to ensure that the ethanol content reaches 60%, standing for 24 hours, filtering the supernatant, recovering the ethanol, concentrating to obtain a thick paste with the relative density of 1.30-1.35 (80 ℃), drying in vacuum, and crushing to obtain dry paste powder for later use;
E. d, taking the dry paste powder obtained in the step D and the fine powder of the leech, the centipede and the scorpion obtained in the step A, adding sugar powder and dextrin, uniformly mixing, adding ethanol, preparing a soft material, sieving, granulating, drying, spraying the volatile oil obtained in the step B, uniformly shaking, drying and sieving to obtain granules; or the following scheme is selected:
and (3) adding cyclodextrin into the volatile oil obtained in the step (B) to prepare inclusion, uniformly mixing the inclusion with the dry paste powder obtained in the step (D), the fine powder of the leech, the centipede and the scorpion obtained in the step (A) and starch, and preparing pills and drying to obtain the pills.
The preparation method of the Mailuoshutong pill comprises the following steps:
A. pulverizing part of Hirudo, Scolopendra, and Scorpio into fine powder;
B. distilling flos Lonicerae, rhizoma Atractylodis, radix scrophulariae, radix Angelicae sinensis, and radix Paeoniae alba with water, and extracting volatile oil; distilling the medicinal liquid and residue;
C. mixing the distillation residue obtained in the step B with radix astragali, cortex Phellodendri, Coicis semen, Glycyrrhrizae radix, the rest Hirudo, Scolopendra and Scorpio, and decocting with water to obtain decoction for use;
D. mixing the distilled liquid medicine obtained in the step B and the decoction obtained in the step C, concentrating, adding alcohol, standing, filtering, recovering ethanol, concentrating, drying, and crushing to obtain fine powder for later use;
E. and (4) preparing the volatile oil in the step (B) into a contained substance, mixing the contained substance with the dry paste powder obtained in the step (D), the fine powder of the leech, the centipede and the scorpion obtained in the step (A) and conventional auxiliary materials, and preparing pills.
Specifically, the preparation method of the Mailuoshutong pill comprises the following steps:
A. 1/2 pulverizing Hirudo, Scolopendra, and Scorpio into fine powder;
B. soaking flos Lonicerae, rhizoma Atractylodis, radix scrophulariae, radix Angelicae sinensis, and radix Paeoniae alba in water, distilling, and collecting volatile oil; distilling to obtain medicinal liquid and residue;
C. b, uniformly mixing the astragalus, the golden cypress, the coix seed, the liquorice, the residual 1/2 formula leech, the centipede and the scorpion with the distilled medicine residues obtained in the step B, adding water for decocting for 2 times, and mixing decoction for later use, wherein the decoction is 1.5 hours each time;
D. mixing the distilled liquid obtained in the step B and the decoction obtained in the step C, filtering, concentrating the filtrate to obtain a clear paste with a relative density of 1.10-1.20 (80 ℃), adding ethanol to ensure that the ethanol content reaches 60%, standing for 24 hours, filtering the supernatant, recovering the ethanol, concentrating to obtain a clear paste with a relative density of 1.10-1.18 (80 ℃), spray drying, and crushing to obtain dry paste powder for later use;
E. and (3) adding cyclodextrin into the volatile oil obtained in the step (B) to prepare inclusion, uniformly mixing the inclusion with the dry paste powder obtained in the step (D), the fine powder of the leech, the centipede and the scorpion obtained in the step (A) and starch, and preparing pills and drying to obtain the pills.
Animal pharmacological experiments prove that the traditional Chinese medicine composition can effectively inhibit the expression of TUNEL positive cells around hematoma of a cerebral hemorrhage rat and inhibit the expression of TNF-alpha protein of neuron cells around the hematoma, thereby inhibiting apoptosis and relieving secondary cerebral tissue injury of the cerebral hemorrhage; the rat ethology of cerebral hemorrhage and apoplexy can be effectively improved, and the nerve function after cerebral hemorrhage is improved; can significantly increase blood flow of cerebral cortex of rat, reduce edema degree of cerebral tissue, and reduce cerebral infarction range, and has effects of protecting cerebral tissue of ischemic apoplexy rat, and improving blood stasis symptom.
The invention is further illustrated by the following examples, which should be properly understood: the examples of the present invention are merely illustrative and not restrictive, and therefore, the present invention may be modified in a simple manner without departing from the scope of the invention as claimed.
Example 1
Prescription:
leech 180g, scorpion 100g and centipede 40 g
Chinese angelica root 100g scrophularia root 700 g coix seed 300g
400 g of phellodendron bark, 300g of honeysuckle flower and 700 g of astragalus root
Licorice root 50g atractylodes rhizome 400 g white peony root 100g
The preparation method comprises the following steps:
A. 1/2 pulverizing Hirudo, Scolopendra, and Scorpio into fine powder;
B. soaking flos Lonicerae, rhizoma Atractylodis, radix scrophulariae, radix Angelicae sinensis, and radix Paeoniae alba in water, distilling, and collecting volatile oil; distilling to obtain medicinal liquid and residue;
C. b, uniformly mixing the astragalus, the golden cypress, the coix seed, the liquorice, the residual 1/2 formula leech, the centipede and the scorpion with the distilled medicine residues obtained in the step B, adding water for decocting for 2 times, and mixing decoction for later use, wherein the decoction is 1.5 hours each time;
D. mixing the distilled liquid medicine obtained in the step B and the decoction obtained in the step C, filtering, concentrating the filtrate to obtain clear paste with the relative density of 1.10-1.20 (80 ℃), adding ethanol to ensure that the ethanol content reaches 60%, standing for 24 hours, filtering the supernatant, recovering the ethanol, concentrating to obtain clear paste with the relative density of 1.10-1.18 (80 ℃) or thick paste with the relative density of 1.30-1.35 (80 ℃), drying, and crushing to obtain dry paste powder for later use;
E. and D, taking the dry paste powder obtained in the step D and the fine powder of the leech, the centipede and the scorpion obtained in the step A, adding sugar powder and dextrin, uniformly mixing, adding ethanol, preparing a soft material, sieving, granulating, drying, spraying the volatile oil obtained in the step B, uniformly shaking, drying and sieving to obtain 1000g of granules.
Example 2
Prescription:
leech 280 g scorpion 50g centipede 10g
400 g of angelica sinensis, 300g of figwort root and 700 g of coix seed
Cortex Phellodendri 100g flos Lonicerae 700 g radix astragali g
100g of liquorice, 100g of rhizoma atractylodis, and 400 g of white paeony root.
The preparation method comprises the following steps: the same as in example 1.
Example 3
Leech 280 g scorpion 50g centipede 40 g
200g of angelica sinensis, 550g of figwort root, 550g of coix seed and 450g of coix seed
300g of golden cypress, 450g of honeysuckle flower and 550g of astragalus root
Licorice root 100g atractylodes rhizome 200g white peony root 280 g
The preparation method comprises the following steps: the same as in example 1.
Example 4
Leech 180g, scorpion 100g and centipede 10g
300g of Chinese angelica, 450g of figwort root and 550g of coix seed
200g of phellodendron bark, 550g of honeysuckle flower, 450g of astragalus root
Licorice root 50g atractylodes rhizome 300g white peony root 200g
The preparation method comprises the following steps: the same as in example 1.
Example 5
Leech 250g, scorpion 83g, centipede 20g
250g of Chinese angelica, 500g of figwort root and 500g of coix seed
Cortex Phellodendri 250g flos Lonicerae 500g radix astragali 500g
Licorice root, radix Glycyrrhizae 83g
The preparation method comprises the following steps: the same as in example 1.
Example 6
Leech 250g, scorpion 83g, centipede 20g
250g of Chinese angelica, 500g of figwort root and 500g of coix seed
Cortex Phellodendri 250g flos Lonicerae 500g radix astragali 500g
Licorice root 83g atractylodes rhizome 250g white peony root 250g
The preparation method comprises the following steps: the same as in example 1.
Example 7
Leech 417g scorpion 138g centipede 33g
Angelica sinensis 417g radix scrophulariae 833g Coix seed 833g
Phellodendron bark 417g honeysuckle 833g astragalus root 833g
138g of liquorice, 417g of rhizoma atractylodis and 417g of white paeony root
The preparation method comprises the following steps:
A. 1/2 pulverizing Hirudo, Scolopendra, and Scorpio into fine powder;
B. soaking flos Lonicerae, rhizoma Atractylodis, radix scrophulariae, radix Angelicae sinensis, and radix Paeoniae alba in water, distilling, and collecting volatile oil; distilling to obtain medicinal liquid and residue;
C. b, uniformly mixing the astragalus, the golden cypress, the coix seed, the liquorice, the residual 1/2 formula leech, the centipede and the scorpion with the distilled medicine residues obtained in the step B, adding water for decocting for 2 times, and mixing decoction for later use, wherein the decoction is 1.5 hours each time;
D. mixing the distilled liquid medicine obtained in the step B and the decoction obtained in the step C, filtering, concentrating the filtrate to obtain clear paste with the relative density of 1.10-1.20 (80 ℃), adding ethanol to ensure that the ethanol content reaches 60%, standing for 24 hours, filtering the supernatant, recovering the ethanol, concentrating to obtain clear paste with the relative density of 1.10-1.18 (80 ℃) or thick paste with the relative density of 1.30-1.35 (80 ℃), drying, and crushing to obtain dry paste powder for later use;
E. and (3) adding cyclodextrin into the volatile oil obtained in the step (B) to prepare inclusion, uniformly mixing the inclusion with the dry paste powder obtained in the step (D), the fine powder of the leech, the centipede and the scorpion obtained in the step (A) and starch, and preparing pills and drying to obtain 1000g of pills.
Comparative example 1
Leech 250g, scorpion 30 g, centipede 50g
400 g of Chinese angelica, 400 g of figwort root, 400 g of coix seed and 350 g of coix seed
Cortex Phellodendri 250g flos Lonicerae 300g radix astragali 500g
Licorice root 85 g atractylodes rhizome 400 g white peony root 300g
The preparation method comprises the following steps: the same as in example 1.
Comparative example 2
Leech 250g, scorpion 90 g, centipede 30 g
250g of Chinese angelica, 520 g of rehmannia root and 500g of coix seed
Cortex Phellodendri 250g flos Lonicerae 500g radix astragali 500g
Licorice root 90 g atractylodes rhizome 220 g gastrodia tuber 220 g
The preparation method comprises the following steps:
A. 1/2 pulverizing Hirudo, Scolopendra, and Scorpio into fine powder;
B. taking honeysuckle, rhizoma atractylodis, rehmannia, angelica and rhizoma gastrodiae according to the prescription amount, soaking in water, distilling and extracting, and collecting volatile oil for later use; distilling to obtain medicinal liquid and residue;
C. b, uniformly mixing the astragalus, the golden cypress, the coix seed, the liquorice, the residual 1/2 formula leech, the centipede and the scorpion with the distilled medicine residues obtained in the step B, adding water for decocting for 2 times, and mixing decoction for later use, wherein the decoction is 1.5 hours each time;
D. mixing the distilled liquid medicine obtained in the step B and the decoction obtained in the step C, filtering, concentrating the filtrate to obtain clear paste with the relative density of 1.10-1.20 (80 ℃), adding ethanol to ensure that the ethanol content reaches 60%, standing for 24 hours, filtering the supernatant, recovering the ethanol, concentrating to obtain clear paste with the relative density of 1.10-1.18 (80 ℃) or thick paste with the relative density of 1.30-1.35 (80 ℃), drying, and crushing to obtain dry paste powder for later use;
E. and D, taking the dry paste powder obtained in the step D and the fine powder of the leech, the centipede and the scorpion obtained in the step A, adding sugar powder and dextrin, uniformly mixing, adding ethanol, preparing a soft material, sieving, granulating, drying, spraying the volatile oil obtained in the step B, uniformly shaking, drying and sieving to obtain 1000g of granules.
Comparative example 3
Leech 250g, scorpion 90 g, centipede 30 g
80g of Chinese angelica, 280 g of figwort and 280 g of poria cocos
500g of phellodendron bark, 750 g of forsythia fruit, 280 g of astragalus root
Licorice root 90 g white atractylodes rhizome 80g white peony root 220 g
The preparation method comprises the following steps:
A. 1/2 pulverizing Hirudo, Scolopendra, and Scorpio into fine powder;
B. taking fructus forsythiae, bighead atractylodes rhizome, figwort root, Chinese angelica and white paeony root according to the prescription amount, soaking in water, distilling and extracting, and collecting volatile oil for later use; distilling to obtain medicinal liquid and residue;
C. uniformly mixing the astragalus, the golden cypress, the poria cocos, the liquorice, the residual 1/2 formula leech, the centipede and the scorpion with the distilled medicine residues obtained in the step B, adding water, decocting for 2 times, and each time lasts for 1.5 hours, and combining decoction for later use;
D. mixing the distilled liquid medicine obtained in the step B and the decoction obtained in the step C, filtering, concentrating the filtrate to obtain clear paste with the relative density of 1.10-1.20 (80 ℃), adding ethanol to ensure that the ethanol content reaches 60%, standing for 24 hours, filtering the supernatant, recovering the ethanol, concentrating to obtain clear paste with the relative density of 1.10-1.18 (80 ℃) or thick paste with the relative density of 1.30-1.35 (80 ℃), drying, and crushing to obtain dry paste powder for later use;
E. and D, taking the dry paste powder obtained in the step D and the fine powder of the leech, the centipede and the scorpion obtained in the step A, adding sugar powder and dextrin, uniformly mixing, adding ethanol, preparing a soft material, sieving, granulating, drying, spraying the volatile oil obtained in the step B, uniformly shaking, drying and sieving to obtain 1000g of granules.
Pharmacological test of animals
The traditional Chinese medicine composition has the effects on the nerve function and the brain tissue apoptosis of a rat suffering from hemorrhagic stroke
1. Animal(s) production
SD rat, age of mouse 12-15 months, license number: SCXK (Shandong) 20140007, supplied by Jinanpunyue laboratory animal Breeding Co., Ltd. Rats were returned to the house and acclimatized for one week.
2. Medicine
The invention comprises the following steps: granules obtained in example 5 and example 6 and pellets obtained in example 7;
comparative example: granules obtained in comparative example 1;
positive control drug: naoxuekang capsule for treating cerebral hemorrhage.
3. Dosage to be administered
Example 5: 5.4 g/kg.d-1；
Example 6: 5.4 g/kg.d-1；
Example 7: 3.24 g/kg.d-1；
Comparative example 1: 5.4 g/kg.d-1；
Positive control drug: 0.04 g/kg.d-1。
4. Cerebral hemorrhagic apoplexy rat model
Molding: rats are anesthetized by intraperitoneal injection of 10% chloral hydrate (400 mg/kg), the rats are fixed in a brain stereotaxic apparatus in a prone position, a front chimney is taken as the center, a small hole with the diameter of 1mm is drilled forwards at the position of 0.2mm and rightwards at the position of 3.0mm by a dental drill, 50 mu L of blood is extracted from the right femoral artery by a microsyringe (heparin by a needle tube), a needle is immediately inserted along the drilled hole on the stereotaxic apparatus at the depth of 6.0mm (namely the tail nucleus position), the blood is slowly injected, the needle is left for 10min after the injection, then the needle head is slowly withdrawn, the drilled hole is sealed by bone wax, the incision is sutured, and 40 ten thousand units of penicillin is injected into the abdominal cavity for conventional anti-infection.
False operation: the procedure was followed, but only the needle was inserted and no blood was injected.
5. Grouping and administration of drugs
Grouping: rats successfully molded were divided into a model group, a group I (i.e., example 5) to be administered, a group II (i.e., example 6) to be administered, a group III (i.e., example 7) to be administered, a group comparative (i.e., comparative example 1), and a positive control group (i.e., positive control group). Taking another normal rat as a blank group; sham operated rats, sham operated group.
Administration: 10ml of corresponding medicines are respectively given to each administration group, the control group and the positive control group; blank, model, and sham groups were given equal volumes of distilled water. The administration was continued for 7 days by gavage once a day.
6. Detection and results
6.1 behavioural assay: the method is characterized in that an improved neuroethology scoring method is adopted to score three behaviors of rat rotation reaction, forepaw holding power and cross bar walking, wherein each behavior is divided into 0, 1, 2, 3 and 4, 0 is divided into normal, and 4 is divided into serious injury. Rats were scored for post-operative and post-administration treatment neuro-behavior, respectively.
6.2 Biochemical assays
Sample preparation: after 24h administration on day 1, day 3 and day 7, 8 rats were sacrificed, brain tissue was removed under cardiac perfusion, dehydrated by conventional alcohol gradient and embedded in paraffin.
And (3) detecting cell apoptosis: detecting by using a nick end labeling method mediated by terminal deoxyribose nucleic acid transferase, observing a sample under a microscope after processing, counting the proportion of positive cells, and taking the average value.
TNF-alpha protein detection method: and (3) detecting by using an immunohistochemical streptomyces avidin-peroxidase method.
6.3 results
6.3.1 neurological Scoring
As can be seen from Table 1, the neurological scores of 1 st, 3 rd and 7d of each group are lower than those of the model group, the score of the administration group of the invention is basically not different from that of the positive control group or is better than that of the positive control group, but the score of the invention is lower than that of the control group; the neurobehavioral score of the rat on the 7 th day is obviously lower than that of the rat on the 1 st day, and the results show that the traditional Chinese medicine composition can effectively improve the neurobehavioral of the rat with the cerebral hemorrhagic stroke.
6.3.2 proportion of apoptosis
TUNEL positive cells were occasionally observed in each field of brain tissue in the blank and sham groups, and the remaining TUNEL positive cells were predominantly present in the peri-hematoma region.
As can be seen from table 2, the percentage of TUNEL-positive cells was lower in each group of rats on days 1, 3 and 7 compared to the model group; the proportion of TUNEL positive cells of the administration group is obviously lower than that of a control group, and compared with the positive control group, the TUNEL positive cells of the administration group have no difference basically or are lower than that of the positive control group; the proportion of TUNEL positive cells on the 7 th day is obviously lower than that on the 1 st day, and the results indicate that the traditional Chinese medicine composition can inhibit the expression of the TUNEL positive cells around the hematoma of the cerebral hemorrhagic stroke rat.
6.3.2 TNF-alpha protein expression
TNF-alpha protein positive is a brownish yellow particle, is mainly located in cytoplasm, and is mainly used for staining the cytoplasm of nerve cells and glial cells in brain tissues around hematoma.
As can be seen from Table 3, the TNF-. alpha.positive cells were found to be lower in the rats of each group on days 1, 3 and 7, respectively, as compared with the model group; the TNF-alpha positive cell proportion of the administration group is obviously lower than that of the control group, and compared with the positive control group, the TNF-alpha positive cell proportion is basically not different or lower than that of the positive control group; and the proportion of TNF-. alpha.positive cells on day 7 was significantly lower than on day 1. The results indicate that the traditional Chinese medicine composition can inhibit the expression of peripheral neuron cell TNF-alpha protein of cerebral hemorrhage rats.
The apoptosis mechanism participates in brain tissue injury secondary to cerebral hemorrhage, the apoptosis-causing medium is TNF-alpha and the like, the expression of TNF-alpha receptor is up-regulated during cerebral hemorrhage, the over-expression of TNF-alpha has neurotoxicity, and TNF-alpha plays an important role in brain immunity and inflammation activities, can influence the activity of vascular endothelial cells and has a regulating effect on the vascular inflammatory process, and can promote the expression of adhesion molecules in the endothelial cells to cause leukocyte adhesion. Aggregate and migrate from the capillaries to the brain, thereby playing a key trigger role in secondary brain injury.
The traditional Chinese medicine composition can effectively inhibit the expression of TUNEL positive cells around hematoma of a cerebral hemorrhage rat and inhibit the expression of TNF-alpha protein of neuron cells around the hematoma, thereby inhibiting apoptosis and relieving secondary cerebral tissue injury of the cerebral hemorrhage; in addition, the traditional Chinese medicine composition can also effectively improve the behaviourology of the cerebral hemorrhage apoplexy rats and improve the nerve function after cerebral hemorrhage.
Secondly, the traditional Chinese medicine composition has the influence on cerebral cortex blood flow and cerebral infarction volume of rats with ischemic cerebral apoplexy
1. Animal(s) production
SD rat, age of mouse 12-15 months, license number: SCXK (Shandong) 20140007, supplied by Jinanpunyue laboratory animal Breeding Co., Ltd. Rats were returned to the house and acclimatized for one week.
2. Medicine
The invention comprises the following steps: granules obtained in example 5 and example 6 and pellets obtained in example 7;
comparative example: granules obtained in comparative example 2;
positive control drug: naoshangtong capsule.
3. Dosage to be administered
Example 5: 5.4 g/kg.d-1；
Example 6: 5.4 g/kg.d-1；
Example 7: 3.24 g/kg.d-1；
Comparative example 2 group: 5.4 g/kg.d-1；
Positive control drug: 0.33 g/kg.d-1。
4. Grouping, administering and moulding
Grouping: rats were randomly divided into a sham-operated group, a model group, a comparative example group (i.e., comparative example 2), a positive control group (i.e., positive control group), a drug administration group i (i.e., example 5), a drug administration group ii (i.e., example 6), and a drug administration group iii (i.e., example 7).
Administration: the administration group, the comparative example group and the positive control group are respectively administered with corresponding liquid medicine by intragastric administration, the sham operation group and the model group are administered with distilled water with the same amount by intragastric administration, the continuous administration is carried out for 7 days, and the model is made after the last administration is carried out for 1 h.
Molding: a photochemistry induced focal cerebral infarction model is used as a model of ischemic stroke, and modeling is started after 1 hour of last administration.
The model group, the administration groups, the control group and the positive control group are anesthetized by 10 percent chloral hydrate (0.35 g/kg of body mass), the stereotaxic instrument is fixed on the head of the rat, after the routine disinfection, the rat is separated along the median incision of the head until the complete skull is exposed, the bone window with the diameter of about 6mm is drilled by a dental flat drill by taking the right side of the sagittal suture and the 3mm behind the coronal suture as the center, the superficial bone plate and the marrow layer of the skull are removed, and the lower bone plate and the dura mater are reserved. Slowly injecting 5% rose bengal 80mg/kg body mass via femoral vein for 1min, irradiating bone window with cold light source for 20min after injection is finished for 5min, and detecting heart rate and respiratory rate of rat during illumination period; the operation of injecting normal saline into femoral vein of the sham operation group is the same as the above molding mode.
5. Detection of
5.1 measurement of Water content in brain tissue
After 24h of molding, 8 rats in each group were anesthetized and decapitated, brains were taken out in ice bath, the olfactory bulbs and the lower brainstem were removed, the pia mater was removed, the brain surface was washed with pre-cooled physiological saline, water was absorbed by filter paper, stored at-20 ℃ for 5min, taken out, vertically divided into left and right halves from the sagittal suture of the brain with a sharp blade, weighed, respectively, and the wet weight was recorded. And quickly putting the weighed semi-brain into a constant-temperature drying oven at 105 ℃, baking for 48 hours, weighing the dry weight, and calculating the water content of the brain tissue and the edema degree of the brain tissue.
5.2 cerebral infarction focus volume determination
After 24h of modeling, 8 rats are anesthetized and decapitated for killing, brains are taken out rapidly, precooled physiological saline is used for flushing residual blood, olfactory bulbs, cerebellum and lower brainstem are removed, brain tissue is frozen at minus 20 ℃ for 20min, coronal section is cut at visual cross, 6 coronal sections are continuously made at intervals of 2mm, the coronal sections are rapidly put into 2% red tetrazole solution and incubated at constant temperature of 37 ℃ for 20min (turning over at 10 min), the coronal section is fixed in 4% paraformaldehyde after dyeing, a scanner scans after 24h to measure the area of an infarct focus, and the area of each brain slice infarct is multiplied by the thickness to be the total infarct volume.
5.3 detection of cerebral blood flow in the center of cerebral cortical infarction
After the model is made, 8 rats are taken from each group for anesthesia, the head is fixed by using a stereotaxic instrument, a laser Doppler probe is fixed on a cantilever crane of a brain stereotaxic instrument, and the probe is contacted with the cerebral cortex by utilizing a bone window opened during the model making so as to detect the cortical blood flow. The measurement point is 3mm on the right side of the sagittal suture and 3mm behind the coronal suture, which is the area of infarction as the illumination center during molding, and the cerebral cortex blood flow is measured, and the detection time is 1, 3, 6, 12 and 24 hours after the infarction.
5.4 results
5.4.1 rat brain tissue Water content and edema degree
As can be seen from Table 4, the water content and edema degree of the brain tissue of each group of rats are obviously lower than those of the model group; the water content and edema degree of the brain tissue of rats in each administration group are obviously lower than those of the control group, and compared with the positive control group, the rat brain tissue is basically not different or lower than that of the positive control group.
5.4.2 cerebral infarct size comparison in rats
As can be seen from Table 5, the cerebral infarction area of each group of rats is significantly lower than that of the model group; the cerebral infarction area of rats in each administration group is obviously smaller than that of a control group, and compared with a positive control group, the cerebral infarction area of rats in each administration group is basically not different or is lower than that of the positive control group.
5.4.3 comparison of cerebral blood flow values (rCBF) in rat infarct centers
As can be seen from Table 6, the cerebral blood flow values of the rats in each group were significantly higher than those of the model group though they had some holes; the cerebral infarction center cerebral blood flow value of rats in each administration group is obviously higher than that of a control group, and compared with a positive control group, the cerebral infarction center cerebral blood flow value of rats in each administration group is basically not different or higher than that of the positive control group.
The results indicate that the traditional Chinese medicine composition can obviously increase the cerebral cortex blood flow of rats, reduce the edema degree of cerebral tissues and reduce the cerebral infarction range, and has the effects of protecting the cerebral tissues of rats with ischemic stroke and improving the blood stasis symptom of the rats.
Claims (10)
- The application of a traditional Chinese medicine composition in preparing a medicine for treating stroke is mainly prepared from leech, scorpion, centipede, Chinese angelica, figwort root, coix seed, golden cypress, honeysuckle, astragalus and liquorice.
- The use of claim 1, wherein the Chinese medicinal composition further comprises atractylodes rhizome and white peony root.
- The use of claim 2, wherein the Chinese medicinal composition is prepared from the following components:leech 180 and 280 portions of scorpion 50-100 portions and centipede 10-40 portions100 parts of angelica sinensis, 400 parts of radix scrophulariae, 300 parts of radix scrophulariae, 700 parts of semen coicis, 300 parts of semen coicis, 700 parts of semen coicis100 parts of phellodendron amurense, 400 parts of honeysuckle, 300 parts of honeysuckle, 700 parts of astragalus, 300 parts of astragalus, 700 parts of astragalus50-100 parts of liquorice, 100 parts of rhizoma atractylodis, 400 parts of white paeony root, 100 parts of white paeony root and 400 parts of white paeony root.
- The use of claim 3, wherein the Chinese medicinal composition is prepared from the following components:leech 180 and 280 portions of scorpion 50-100 portions and centipede 10-40 portions300 parts of 200 parts of radix angelicae sinensis, 550 parts of 450 parts of radix scrophulariae, 550 parts of 450 parts of semen coicis200 parts of phellodendron amurense, 450 parts of honeysuckle, 550 parts of astragalus mongholicus, 450 parts of astragalus membranaceus50-100 parts of liquorice, 200 parts of rhizoma atractylodis, 200 parts of white paeony root and 280 parts of white paeony root;one of the preferable methods is:leech 250 parts, scorpion 83 parts, centipede 20 parts280 parts of Chinese angelica, 530 parts of figwort root, 460 parts of coix seed280 parts of golden cypress, 460 parts of honeysuckle flower, 530 parts of astragalus root83 parts of liquorice, 220 parts of rhizoma atractylodis, 220 parts of white paeony root;the second is preferably:leech 250 parts, scorpion 83 parts, centipede 20 parts250 parts of Chinese angelica, 500 parts of figwort and 500 parts of coix seed250 parts of phellodendron bark, 500 parts of honeysuckle flower and 500 parts of astragalus rootLicorice root 83 weight portions, atractylodes rhizome 250 weight portions, white peony root 250 weight portions.
- The use of any one of claims 1-4, wherein said Angelica sinensis (oliv.) Diels is wine-processed Angelica sinensis (oliv.) Diels and said phellodendron amurense (phellodendron amurense Rupr. ex Benth) is salted phellodendron amurense.
- The use of any one of claims 1-4, wherein said Chinese medicinal composition is formulated into a clinically acceptable dosage form; preferably, the dosage form is a tablet, a granule, a capsule or a pill; further preferably, the dosage form is granules or pills; more preferably, the granules are Mailuoshutong granules, and the pills are Mailuoshutong pills.
- Use according to any one of claims 1 to 4, wherein the stroke is a hemorrhagic stroke and/or an ischemic stroke.
- The use according to claim 7, wherein the hemorrhagic stroke is cerebral hemorrhage and/or subarachnoid hemorrhage; the ischemic stroke is cerebral infarction and/or cerebral thrombosis.
- The use as claimed in claim 6, wherein the preparation method of the Mailuoshutong granule is as follows:A. pulverizing part of Hirudo, Scolopendra, and Scorpio into fine powder;B. distilling flos Lonicerae, rhizoma Atractylodis, radix scrophulariae, radix Angelicae sinensis, and radix Paeoniae alba with water, and extracting volatile oil; distilling the medicinal liquid and residue;C. mixing the distillation residue obtained in the step B with radix astragali, cortex Phellodendri, Coicis semen, Glycyrrhrizae radix, the rest Hirudo, Scolopendra and Scorpio, and decocting with water to obtain decoction for use;D. mixing the distilled liquid medicine obtained in the step B and the decoction obtained in the step C, concentrating, adding alcohol, standing, filtering, recovering ethanol, concentrating, drying, and crushing to obtain fine powder for later use;E. and D, mixing the fine powder obtained in the step D and the fine powder obtained in the step A, adding conventional auxiliary materials, granulating, drying, and spraying the volatile oil obtained in the step B to obtain granules.
- The use of claim 6, wherein the preparation method of the Mailuoshutong pill is as follows:A. pulverizing part of Hirudo, Scolopendra, and Scorpio into fine powder;B. distilling flos Lonicerae, rhizoma Atractylodis, radix scrophulariae, radix Angelicae sinensis, and radix Paeoniae alba with water, and extracting volatile oil; distilling the medicinal liquid and residue;C. mixing the distillation residue obtained in the step B with radix astragali, cortex Phellodendri, Coicis semen, Glycyrrhrizae radix, the rest Hirudo, Scolopendra and Scorpio, and decocting with water to obtain decoction for use;D. mixing the distilled liquid medicine obtained in the step B and the decoction obtained in the step C, concentrating, adding alcohol, standing, filtering, recovering ethanol, concentrating, drying, and crushing to obtain fine powder for later use;E. and (4) preparing the volatile oil in the step (B) into a contained substance, mixing the contained substance with the dry paste powder obtained in the step (D), the fine powder of the leech, the centipede and the scorpion obtained in the step (A) and conventional auxiliary materials, and preparing pills.
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