CN117243991A - Application of osmanthus fragrans or extract thereof in preparation of medicines for preventing and/or treating cerebral ischemia injury - Google Patents
Application of osmanthus fragrans or extract thereof in preparation of medicines for preventing and/or treating cerebral ischemia injury Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/63—Oleaceae (Olive family), e.g. jasmine, lilac or ash tree
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
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- Bioinformatics & Cheminformatics (AREA)
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- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
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Abstract
The invention provides an application of osmanthus fragrans or an extract thereof in preparing a medicament for preventing and/or treating cerebral ischemia injury. According to the invention, through a cell model and an animal model of chronic cerebral ischemia injury, the activity of the sweet osmanthus extract for effectively improving the cells lack of sugar and oxygen is proved; restoring cerebral blood flow in the mice after chronic ischemia. The osmanthus fragrans or the extract thereof is prepared into the medicine for preventing and/or treating the chronic cerebral ischemia injury, and has wide market application prospect.
Description
Technical Field
The invention relates to an application of osmanthus fragrans or an extract thereof in preparing a medicament for preventing and/or treating cerebral ischemia injury.
Background
Cerebral ischemia is a common pathological condition of the nervous system. For many years, research on cerebral ischemia has been focused mainly on acute cerebral ischemia, but the understanding of the characteristics, mechanism and prevention of cerebral injury caused by chronic cerebral ischemia is not clear. Chronic cerebral ischemia or cerebral perfusion deficiency is accompanied with cerebral arteriosclerosis, vascular dementia, alzheimer's disease, binswanger's disease, arteriovenous malformations and other cerebral vascular diseases, cognitive function damage is mainly represented in early stage of onset, and finally persistent or progressive cognitive and nerve dysfunction may be caused.
The osmanthus fragrans are a plant belonging to the Oleaceae, and are classified into silver osmanthus, gold osmanthus and red osmanthus according to different colors. The osmanthus fragrans contains various components including iridoids, phenols, lignans and the like, the osmanthus fragrans fragrance is rich in a large amount of aromatic components, and most of the osmanthus fragrans fragrance is terpene substances with antioxidant activity, so that the osmanthus fragrans has high medicinal value. For example, the osmanthus flavone has a remarkable inhibition effect on the in vitro proliferation of human liver cancer Hep G2 cells; the osmanthus fragrans extract can play a good anti-aging role when being acted on skin. However, no research report on treating chronic cerebral ischemia injury by using osmanthus fragrans or an extract thereof exists at present.
Disclosure of Invention
In order to solve the problems, the invention provides an application of osmanthus fragrans or an extract thereof in preparing a medicament for preventing and/or treating cerebral ischemia injury.
Further, the extract is a water, alcohol or ethyl acetate extract of osmanthus fragrans.
Further, the extract is an ethanol extract of osmanthus fragrans.
Still further, the extract is a 95% ethanol extract of osmanthus fragrans.
Further, the osmanthus flower is a flower of the plant luteolin Osmanthus fragrans (thunder.) lour.
Further, the medicament is a medicament for preventing and/or treating chronic cerebral ischemia injury.
Further, the drug is a drug for improving the activity of the hypoxic and ischemic nerve cells.
Still further, the drug is a drug that restores cerebral blood flow.
Further, the medicine is a preparation prepared by taking osmanthus fragrans or an extract thereof as an active ingredient and adding pharmaceutically acceptable auxiliary materials or auxiliary ingredients.
Still further, the formulation is an oral formulation; the oral preparation is granule, solution, pill, paste or tablet; the external preparation is solution, ointment, gel or emulsion.
Further, the preparation is an external preparation; the external preparation is solution, ointment, gel or emulsion.
The application of the osmanthus fragrans or the osmanthus fragrans extract in preparing a medicament for preventing and/or treating cerebral ischemia injury proves that the osmanthus fragrans extract can effectively improve the activity of the cells with the functions of sugar deficiency and oxygen deficiency through a cell model and an animal model of chronic cerebral ischemia injury; restoring cerebral blood flow in the mice after chronic ischemia. The osmanthus fragrans or the extract thereof is prepared into the medicine for preventing and/or treating the chronic cerebral ischemia injury, and has wide market application prospect.
It should be apparent that, in light of the foregoing, various modifications, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.
The above-described aspects of the present invention will be described in further detail below with reference to specific embodiments in the form of examples. It should not be understood that the scope of the above subject matter of the present invention is limited to the following examples only. All techniques implemented based on the above description of the invention are within the scope of the invention.
Detailed Description
Example 1 preparation of Osmanthus fragrans extract
50g of osmanthus fragrans are weighed and put into a 1000mL round-bottom flask, 600mL of 95% ethanol is injected, heating reflux extraction is carried out for 2 hours, and the extract is filtered by a 200-mesh filter cloth, so as to obtain filtrate. Recovering ethanol from the filtrate under reduced pressure, and concentrating until no ethanol smell exists to obtain fluid extract. Transferring the fluid extract into penicillin bottle, pre-freezing at-80deg.C, and lyophilizing to obtain flos Osmanthi Fragrantis ethanol extract.
Example 2 preparation of Osmanthus fragrans extract
50g of osmanthus fragrans are weighed and put into a 1000mL round-bottom flask, 600mL of water is injected, heating reflux extraction is carried out for 2 hours, and the extracting solution is filtered by a 200-mesh filter cloth, so as to obtain filtrate. Concentrating the filtrate under reduced pressure to relative density of 1.05-1.15 to obtain fluid extract. Drying the fluid extract by spray drying to obtain Gui Huashui extract.
EXAMPLE 3 preparation of Osmanthus fragrans extract
50g of osmanthus fragrans are weighed and put into a 1000mL round-bottom flask, 600mL of ethyl acetate is injected, heating reflux extraction is carried out for 2 hours, and the extracting solution is filtered by a 200-mesh filter cloth, so as to obtain filtrate. Recovering ethyl acetate from the filtrate under reduced pressure, and concentrating until no solvent smell exists to obtain fluid extract. Transferring the fluid extract into penicillin bottle, pre-freezing at-80deg.C, and lyophilizing to obtain flos Osmanthi Fragrantis ethyl acetate extract.
Example 4 preparation of Osmanthus fragrans extract-containing tablet
Taking the osmanthus extract prepared in the example 1, respectively adding lactose, sodium carboxymethyl cellulose and sodium carboxymethyl starch, uniformly mixing, granulating, adding magnesium stearate, uniformly mixing, and tabletting to prepare tablets.
Example 5 preparation of Osmanthus fragrans extract-containing pill
Taking the osmanthus fragrans extract prepared in the embodiment 1, adding microcrystalline cellulose, sodium carboxymethyl cellulose and sodium carboxymethyl starch, uniformly mixing, adding 70% ethanol, mixing to prepare a soft material, preparing pills by adopting a plastic method, and drying to obtain the pills.
Example 6 preparation of an emulsion containing Osmanthus extract
Dissolving flos Osmanthi Fragrantis extract prepared in example 1 in water, dispersing, adding edible oil, phospholipid and polyethylene glycol fatty acid ester, and grinding in high pressure homogenizer to obtain emulsion.
EXAMPLE 7 preparation of Osmanthus extract-containing solution
Dissolving and dispersing the osmanthus fragrans extract prepared in the embodiment 1 in water, adding the solubilizing agent Tween 80, uniformly mixing, and finally adding water for dilution to obtain a solution.
Example 8 preparation of gel containing Osmanthus extract
Dissolving and dispersing the osmanthus fragrans extract prepared in the embodiment 1 in water, adding a matrix material poloxamer P407 or hydroxypropyl methylcellulose, uniformly mixing, and finally diluting with water to obtain the gel.
The beneficial effects of the present invention are further illustrated by the following test examples
Test example 1 study of influence of Osmanthus fragrans extract on PC12 cells deficient in sugar and oxygen
1 Experimental method
Taking PC12 cells in logarithmic growth phase, subjecting to digestion and centrifugation (800 rpm/min,5 min), inoculating into 96-well plate (density 1×10) 5 mu.L/well, 100. Mu.L/well), for 24h. The supernatant was discarded and the cells were divided into: normal group, model group, extract group with different concentration. Complete medium, 120. Mu.L/well, was added to the normal group. The other groups were each first charged with 0.4mmol/L cobalt chloride, 100. Mu.L/well, prepared in sugarless DMEM medium. In addition, the model group was further added with 20. Mu.L of sugar-free medium to fill up the volume, and 20. Mu.L of the medicinal liquid was further added to each of the administration groups (the concentrations of the extracts were calculated from the crude osmanthus fragrans, and the final concentrations are shown in tables 1 to 3). Each group had 6 multiple wells in an incubator (37 ℃ C., 5% CO) 2 ) The MTT method was used to detect absorbance values after further incubation for 24h.
2 experimental results
2.1 protective action of Osmanthus fragrans extract on sugar-deficient and anoxic nerve cells
And (3) incubating and culturing for 24 hours by adopting 0.4mmol/L cobalt chloride combined sugar-free culture medium, establishing an in-vitro glucose-lack and oxygen-lack PC12 cell model to simulate chronic cerebral ischemia injury, detecting cell activity by an MTT method, and evaluating cerebral ischemia protection effect of the osmanthus fragrans flower extract, wherein specific statistical results are shown in tables 1-3.
As can be seen from the results in Table 1, the alcohol extract concentration (0.15-1.5 mg/ml) has the effect of promoting the in vitro survival of the sugar-deficient and anoxic cells (P < 0.01) when compared with the model group.
As can be seen from the results in Table 2, the aqueous extract concentration (0.6-1.2 mg/ml) has the effect of promoting the in vitro survival of the sugar-deficient and anoxic cells (P <0.05 or 0.01) when compared with the model group.
As can be seen from the results in Table 3, the ethyl acetate extract concentration (0.3-0.9 mg/ml) had the effect of promoting the in vitro survival of the sugar-deficient and oxygen-deficient cells (P <0.05 or 0.01) when compared with the model group.
TABLE 1 influence of Osmanthus fragrans alcohol extract on cell survival due to lack of sugar and oxygenn＝6)
Note that: ratio to model group P <0.05, P <0.01
TABLE 2 influence of Osmanthus fragrans aqueous extract on cell survival due to lack of sugar and oxygenn＝6)
Note that: ratio to model group P <0.05, P <0.01
TABLE 3 influence of Osmanthus fragrans ethyl acetate extract on cell survival due to lack of sugar and oxygenn＝6)
Note that: ratio to model group P <0.05, P <0.01
Test example 2 animal experiment efficacy verification of Osmanthus fragrans alcohol extract
1. Method of
1.1 replication of the mouse model of chronic cerebral ischemia
A chronic cerebral ischemia mouse model is established by adopting a right common carotid artery ligation method, and the mice are fasted for 12 hours before operation and weighed. The 10% chloral hydrate is used for intraperitoneal injection to anesthetize the mice, the mice are fixed in the supine position, and the neck is prepared for skin and disinfection. A small incision of about 1-2cm was made along the midline of the neck with scissors, and the fascia muscles were separated to find the left common carotid artery. The distal and proximal ends were then ligated with silk threads 0-4, cutting the right common carotid artery from the middle. Finally, the neck incision is sutured with surgical threads, and the prosthetic operation group only separates the common carotid artery and does not ligate. Penicillin sodium is injected into muscle for 3 days after operation to prevent infection.
1.2 grouping and administration
Mice that were successfully modeled were randomly divided into three groups, the model group, the sham operation group, and the dosing group. The alcohol extract of osmanthus flower prepared in example 1 is administered to a dosing group, the dosage of the alcohol extract is calculated to be 780mg/kg according to the daily 6g of crude osmanthus flower drug used by a person and the body surface area coefficient of a mouse to be 0.0026, so that the alcohol extract is administered by stomach irrigation according to 0.1ml/10 g; the model and sham groups were given equal volumes of saline. The cerebral blood flow on the left side and the right side is monitored by a laser speckle blood flow meter respectively after 30 days of postoperative immediate and continuous administration, the difference value is calculated, and the blood flow recovery rate of each group after administration is calculated.
Differential cerebral blood flow on both sides = left cerebral blood flow-right cerebral blood flow
Right side blood flow recovery rate= (postoperative bilateral cerebral blood flow difference-administration of cerebral blood flow difference of 30 days) postoperative bilateral cerebral blood flow difference
2. Results
The specific statistical results are shown in Table 4 below. Compared with the model group, the blood flow difference value at two sides of the osmanthus fragrans extract administration group is obviously reduced, the blood flow recovery rate of the osmanthus fragrans extract administration group is obviously improved (P is less than 0.05), and the drug is helpful for cerebral blood flow recovery after chronic ischemia so as to protect ischemic nerve cells.
Table 4 groups were treated with chronic cerebral ischemia miceCerebral blood flow change after administration for 30 daysn＝8)
Note that: p <0.05, P <0.01 compared to model group.
In conclusion, the cell model and the animal model of chronic cerebral ischemia injury prove that the osmanthus fragrans extract can effectively improve the activity of the cells with the functions of lack of sugar and oxygen; restoring cerebral blood flow in the mice after chronic ischemia. The osmanthus fragrans or the extract thereof is prepared into the medicine for preventing and/or treating the chronic cerebral ischemia injury, and has wide market application prospect.
Claims (10)
1. Application of flos Osmanthi Fragrantis or its extract in preparing medicine for preventing and/or treating cerebral ischemia injury is provided.
2. The use according to claim 1, wherein the extract is a water, alcohol or ethyl acetate extract of osmanthus fragrans.
3. Use according to claim 2, characterized in that the extract is an ethanol extract of osmanthus fragrans, preferably a 95% ethanol extract.
4. The use according to claim 1, characterized in that the osmanthus fragrans is a flower of the plant luteolin Osmanthus fragrans (thunder.) lour.
5. The use according to claim 1, wherein the medicament is a medicament for the prevention and/or treatment of chronic cerebral ischemic injury.
6. The use according to claim 5, wherein the medicament is a medicament for restoring cerebral blood flow.
7. The use according to claim 6, wherein the medicament is a medicament for increasing the activity of hypoxic and sugar-deficient nerve cells.
8. The use according to any one of claims 1 to 7, wherein the medicament is a preparation prepared by adding pharmaceutically acceptable auxiliary materials or auxiliary ingredients to osmanthus fragrans or an extract thereof as an active ingredient.
9. The use according to claim 8, wherein the formulation is an oral formulation; the oral preparation is granule, solution, pill, paste or tablet; the external preparation is solution, ointment, gel or emulsion.
10. The use according to claim 9, wherein the formulation is an external formulation; the external preparation is solution, ointment, gel or emulsion.
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