CN111184837B - Pharmaceutical composition for preventing and treating cerebral ischemia injury and application thereof - Google Patents

Abstract

The invention discloses a pharmaceutical composition for preventing and treating cerebral ischemia injury and application thereof, wherein the composition comprises the following components: 48-144 parts of myrobalan pulp, 48-144 parts of terminalia chebula pulp, 48-144 parts of emblic leafflower fruit, 48-144 parts of Chinese olive, 24-72 parts of kadsura root-bark, 24-72 parts of Eurasian fossilizid, 24-72 parts of semen cuscutae, 24-72 parts of lavender, 14.4-43.2 parts of Alihong, 4.8-14.4 parts of Tilia cordata, 4.8-14.4 parts of Nardostachys chinensis, 4.8-14.4 parts of cardamom, 4.8-14.4 parts of frankincense, 4.8-14.4 parts of hedyotis diffusa, 4.8-14.4 parts of dried orange peel, 4.8-14.4 parts of rheum officinale, 4.8-14.4 parts of saffron, 24-72 parts of rose and 24-72 parts of liquorice extract. Experiments prove that the pharmaceutical composition of the invention has significant therapeutic effect in a rat permanent focal cerebral ischemia model.

Description

Pharmaceutical composition for preventing and treating cerebral ischemia injury and application thereof

Technical Field

The invention relates to a pharmaceutical composition and application thereof, in particular to a pharmaceutical composition for preventing and treating cerebral ischemic injury and application thereof in preparing a medicament for preventing and treating cerebral ischemic injury.

Background

Stroke is an acute cerebrovascular disease, a group of diseases that causes damage to brain tissue due to sudden rupture of cerebral vessels or failure of blood to flow into the brain due to vessel occlusion, including ischemic stroke and hemorrhagic stroke. The incidence of ischemic stroke is higher than hemorrhagic stroke, accounts for about 70 percent of all stroke, is common disease and frequently encountered diseases of middle-aged and elderly people, has the characteristics of high fatality rate, high disability rate and the like, and most survivors leave serious neurological impairment symptoms, thereby seriously threatening the health and the life quality of human beings. The occlusion and stenosis of internal carotid and vertebral arteries can cause the sudden reduction or stop of local blood supply of brain tissue, which causes the cerebral tissue ischemia and hypoxia in the blood supply area of the blood vessel to lead to the necrosis and softening of the brain tissue, and is accompanied with clinical symptoms and signs of corresponding parts, such as hemiplegia, aphasia and other symptoms of nerve function loss. At present, the main methods for treating cerebral ischemia in western medicine include intravenous thrombolysis and mechanical thrombus removal, but usually along with nerve injury and dysfunction caused by ischemia, a neuroprotective medicament capable of effectively preventing and treating cerebral ischemia injury needs to be found. To date, there is no published report on the treatment of cerebral infarction by the present pharmaceutical composition.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provide a pharmaceutical composition for preventing and treating cerebral ischemic injury.

The second purpose of the invention is to provide the application of the pharmaceutical composition for preventing and treating the cerebral ischemic injury.

The technical scheme of the invention is summarized as follows:

a pharmaceutical composition for preventing and treating cerebral ischemia injury is prepared from the following components in parts by mass: 48-144 parts of myrobalan pulp, 48-144 parts of terminalia chebula pulp, 48-144 parts of emblic leafflower fruit, 48-144 parts of Chinese olive, 24-72 parts of kadsura root-bark, 24-72 parts of Eurasian fossilizid, 24-72 parts of semen cuscutae, 24-72 parts of lavender, 14.4-43.2 parts of Alihong, 4.8-14.4 parts of Tilia cordata, 4.8-14.4 parts of Nardostachys chinensis, 4.8-14.4 parts of cardamom, 4.8-14.4 parts of frankincense, 4.8-14.4 parts of hedyotis diffusa, 4.8-14.4 parts of dried orange peel, 4.8-14.4 parts of rheum officinale, 4.8-14.4 parts of saffron, 24-72 parts of rose and 24-72 parts of liquorice extract.

The medicine composition is prepared by the following steps:

pulverizing the fomes officinalis, the illicium ferruginea, the Nardostachys chinensis, the cardamom, the frankincense, the hedyotis diffusa, the dried orange peel and the rhubarb into fine powder, and sieving the fine powder by a No. 5 sieve to obtain a mixture 1; grinding stigma croci Sativi, and mixing with mixture 1 to obtain mixture 2; decocting fructus Chebulae, fructus Terminaliae Billericae, fructus Phyllanthi, fructus Chebulae, radix Seu caulis Kadsurae Heteroclitae, Os Draconis Eurasianae, semen Cuscutae, Lavender and flos Rosae Rugosae in water, mixing filtrates, concentrating to relative density of 1.10-1.20 at 65 deg.C, adding Glycyrrhrizae radix extract, and mixing with mixture 2.

Adding water for decocting for 1-3 times, wherein the amount of water added is 8-12 times of the total weight of fructus Chebulae, fructus Terminaliae Billericae, fructus Phyllanthi, fructus Chebulae, radix Seu caulis Kadsurae Heteroclitae, Os Draconis Eurasiabell, semen Cuscutae, Lavender and flos Rosae Rugosae.

The application of the pharmaceutical composition in preparing medicines for preventing and treating cerebral ischemia injury.

The invention has the beneficial effects that:

animal experiments prove that the pharmaceutical composition for preventing and treating cerebral ischemic injury plays a significant role in treating a permanent focal cerebral ischemia model of a rat.

Drawings

FIG. 1 is a graph showing the effect of a pharmaceutical composition of the present invention on the percent of cerebral infarction in rats in the model of permanent focal cerebral ischemia (pMCAO) in rats; compared with the sham operation group^**P＜001; compared with model group^##P is less than 0.01; compared with the group of blood-nourishing and brain-refreshing granules^△P＜0.05。

FIG. 2 is a TTC staining pattern of brain sections.

Detailed Description

The present invention is further illustrated but not limited in any way by the following examples, and any modifications or alterations based on the present invention are within the scope of the present invention.

The Glycyrrhrizae radix extract is prepared from Glycyrrhrizae radix. Conforms to the Chinese pharmacopoeia (2015 edition I).

Example 1

A pharmaceutical composition for preventing and treating cerebral ischemia injury is prepared from the following components: 48g of myrobalan pulp, 48g of terminalia chebula pulp, 48g of emblic leafflower fruit, 48g of Chinese olive, 24g of kadsura root-bark, 24g of eupolyphaga, 24g of dodder, 24g of lavender, 14.4g of fomes officinalis, 4.8g of cassia twig, 4.8g of rhizoma nardostachyos, 4.8g of cardamom, 4.8g of frankincense, 4.8g of oldenlandia diffusa, 4.8g of dried orange peel, 4.8g of rheum officinale, 4.8g of saffron, 24g of rose and 24g of liquorice extract.

The preparation method of the pharmaceutical composition comprises the following steps:

pulverizing the fomes officinalis, the illicium ferruginea, the Nardostachys chinensis, the cardamom, the frankincense, the hedyotis diffusa, the dried orange peel and the rhubarb into fine powder, and sieving the fine powder by a No. 5 sieve to obtain a mixture 1; grinding stigma croci Sativi, and mixing with mixture 1 to obtain mixture 2; adding water into myrobalan pulp, terminalia chebula pulp, phyllanthus emblica, fructus chebulae immaturus, kadsura coccinea root bark, eurasian dragon's bones, semen cuscutae, lavender and roses for decoction, wherein the amount of the water added for decoction is 8 times of the total mass of the myrobalan pulp, the terminalia chebula pulp, the phyllanthus emblica, the terminalia chebula, the kadsura coccinea root bark, the eurasian dragon's bones, the semen cuscutae, the lavender and the roses, the decoction time is 1h, the decoction is carried out for 3 times, the filtrates are combined, the filtrate is concentrated to the relative density of 1.10 at 65 ℃, the liquorice extract is added, and the mixture is uniformly mixed with the mixture 2 to obtain the traditional Chinese medicine.

Example 2

A pharmaceutical composition for preventing and treating cerebral ischemia injury is prepared from the following components: 144g of myrobalan pulp, 144g of terminalia chebula pulp, 144g of emblic leafflower fruit, 144g of Chinese olive, 72g of kadsura root-bark, 72g of eupolyphaga, 72g of Chinese dodder seed, 72g of lavender, 43.2g of fomes officinalis, 14.4g of cassia twig, 14.4g of rhizoma nardostachyos, 14.4g of cardamom, 14.4g of frankincense, 14.4g of hedyotis diffusa, 14.4g of dried orange peel, 14.4g of rheum officinale, 14.4g of saffron, 72g of rose and 72g of liquorice extract.

The preparation method of the pharmaceutical composition comprises the following steps:

pulverizing the fomes officinalis, the illicium ferruginea, the Nardostachys chinensis, the cardamom, the frankincense, the hedyotis diffusa, the dried orange peel and the rhubarb into fine powder, and sieving the fine powder by a No. 5 sieve to obtain a mixture 1; grinding stigma croci Sativi, and mixing with mixture 1 to obtain mixture 2; adding water into myrobalan pulp, terminalia chebula pulp, phyllanthus emblica, fructus chebulae immaturus, kadsura coccinea root bark, eurasian dragon's bones, semen cuscutae, lavender and roses for decoction, wherein the amount of the water added for decoction is 12 times of the total mass of the myrobalan pulp, the terminalia chebula pulp, the phyllanthus emblica, the terminalia chebula, the kadsura coccinea root bark, the eurasian dragon's bones, the semen cuscutae, the lavender and the roses, the decoction time is 3 hours, the decoction is carried out for 1 time, the filtrates are combined, the filtrate is concentrated to the relative density of 1.20 at 65 ℃, the liquorice extract is added, and the mixture is uniformly mixed with the mixture 2 to obtain the traditional Chinese medicine.

Example 3

A pharmaceutical composition for preventing and treating cerebral ischemia injury, and its neuroprotective effect on rats with permanent cerebral ischemia

1. Material

1.1 drug formulation

The administration dosage of the blood-nourishing and brain-refreshing granules for rats is calculated according to 3 times of the daily dosage of adults (60kg), namely 3.75g/kg of the blood-nourishing and brain-refreshing granules; the dosage of the nimodipine rat is calculated according to the daily dosage of an adult (60kg), namely, 0.0125g/kg of nimodipine; the dosage of the drug composition of the invention administered to rats was 1.08g/kg in the low dose group (example 1) and 3.24g/kg in the high dose group (example 1).

1.2 reagents

Chloral hydrate 10% (analytical grade/20120629, department of Tianjin, Mimi Euro Chemicals Co., Ltd.); gentamicin sulfate (20 ten thousand units/020161505, Proteus animal health products Co., Tianjin); physiological saline (batch No. E14030903, Hebei Tian Cheng Yao GmbH)

1.3 Experimental instruments

One hundred thousand balance model XS105 (M0ETTER TOLEDO); a balance of type BS124S (Sartorius, germany); DW-YL270 model refrigerator (Mike Mitsubishi Low temperature science and technology Co., Ltd.); BM-40 type pure water preparation system (Beijing Zhongsheng Luoyuan scientific and technological development Co., Ltd.); micropipettes (Eppendorf, germany); ALLEGRA-64R high speed centrifuge (EECKMAN, USA).

1.4 Experimental animals

SPF SD rat, male, weight 280-.

The experimental animals are purchased and then raised in an experimental animal room of New drug evaluation Limited company of Tianjin pharmaceutical research institute, the light and shade are controlled at intervals of 12h in a common raising room, the temperature is kept at 16-26 ℃, the humidity is kept at 40-70%, and the ventilation frequency is not less than 8 times of fresh air/hour.

2. Test method

Healthy clean SD male rats 60 were selected and randomly divided into 6 groups: the drug composition comprises a sham operation group, a model group, a nimodipine group, a blood-nourishing and brain-refreshing granule group, a low-dose group and a high-dose group (n is 10).

Adaptive feeding for 3 days. Each group was administered prophylactically for 3 days, anesthetized 30 minutes after the last administration, and a permanent rat cerebral infarction model was created by a wire-embolus method after anesthesia. After the rats revive, the rats are subjected to behavioral staging according to the Longa scoring standard, and rats with failed model building are removed. See table 1.

TABLE 1 Longa score criteria

Note: score lowest 0, score highest 4, higher score representing more severe nerve damage

After 24h, weigh and conduct behavioral staging again on Longa scoring scale. After anesthesia with 10% chloral hydrate, blood was taken from the abdominal aorta, and the upper serum was collected by centrifugation and stored at-20 ℃. After blood draw, the cells were washed with PBS: 1 of physiological saline: 150ml of buffer 20 was used to rinse the blood from the heart through systemic circulation. The brain is removed by opening the cranium, stained with TTC, photographed and preserved in formalin.

Percent cerebral infarction was calculated using Image J (version 1.42) Image processing software.

3. Results of the experiment

3.1 neurological deficit scoring results

As shown in fig. 1, the rat neurological score in the sham operation group was 0, and the score in the model group was significantly higher (3 ± 0.93) compared to the sham operation group, indicating that the model was successfully established. Compared with the model group, the two positive medicine groups (the nimodipine group and the blood-nourishing and brain-refreshing granule group) and the low-dose group (example 1) and the high-dose group (example 1) of the medicine composition have the effect of improving the nerve function damage of rats. See table 2.

TABLE 2

Group of	Dosage (g/kg)	Neurological scoring
			Artificial operation group	—	0.00±0.00
Model set	—	3±0.93**
			Blood-nourishing and brain-refreshing granule group	3.75	2.83±0.98
Nimodipine group	0.0125	1.8±0.45#
			Pharmaceutical composition group (Low dose)	1.08	2.2±0.84
Pharmaceutical composition group (high dose)	3.24	1.5±0.58#

(neurological score as mean. + -. variance)

3.2 measurement results of the volume of cerebral infarction foci

As can be seen from fig. 1, compared with the sham group, the cerebral infarction rate of the model group was significantly increased (P <0.01), indicating that the modeling was successful; compared with the model group, the cerebral infarction rates of the blood-nourishing and brain-refreshing granule group, the nimodipine group and the pharmaceutical composition low-dose group and the high-dose group are remarkably reduced (P < 0.01).

As can be seen from FIG. 2, after modeling, the infarct size is significantly reduced after administration of the positive drug (blood-nourishing and brain-refreshing granules and nimodipine) and the pharmaceutical composition of the present invention (low dose and high dose).

Experiments prove that the effect of the pharmaceutical composition for preventing and treating cerebral ischemic injury prepared in example 2 is similar to that of example 1.

4. Conclusion of the study

The pharmaceutical composition can improve the nerve function damage, effectively reduce the cerebral infarction rate and has the nerve protection effect on the brain of a pMCAO rat.

Claims (3)

1. A pharmaceutical composition for preventing and treating cerebral ischemia injury is characterized by being prepared from the following components: 144g of myrobalan pulp, 144g of terminalia chebula pulp, 144g of emblic leafflower fruit, 144g of Chinese olive, 72g of kadsura root-bark, 72g of eupolyphaga, 72g of Chinese dodder seed, 72g of lavender, 43.2g of fomes officinalis, 14.4g of cassia twig, 14.4g of rhizoma nardostachyos, 14.4g of cardamom, 14.4g of frankincense, 14.4g of hedyotis diffusa, 14.4g of dried orange peel, 14.4g of rheum officinale, 14.4g of saffron, 72g of rose and 72g of liquorice extract.

2. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is prepared by the following method:

pulverizing the fomes officinalis, the illicium ferruginea, the Nardostachys chinensis, the cardamom, the frankincense, the hedyotis diffusa, the dried orange peel and the rhubarb into fine powder, and sieving the fine powder by a No. 5 sieve to obtain a mixture 1; grinding stigma croci Sativi, and mixing with mixture 1 to obtain mixture 2; adding water into myrobalan pulp, terminalia chebula pulp, phyllanthus emblica, fructus chebulae immaturus, kadsura coccinea root bark, eurasian dragon's bones, semen cuscutae, lavender and roses for decoction, wherein the amount of the water added for decoction is 12 times of the total mass of the myrobalan pulp, the terminalia chebula pulp, the phyllanthus emblica, the terminalia chebula, the kadsura coccinea root bark, the eurasian dragon's bones, the semen cuscutae, the lavender and the roses, the decoction time is 3 hours, the decoction is carried out for 1 time, the filtrates are combined, the filtrate is concentrated to the relative density of 1.20 at 65 ℃, the liquorice extract is added, and the mixture is uniformly mixed with the mixture 2 to obtain the traditional Chinese medicine.

3. Use of a pharmaceutical composition according to any one of claims 1 or 2 for the preparation of a medicament for the prevention and treatment of cerebral ischemic injury.

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