CN111803546A - Application of composition of strawberries with centella asiatica - Google Patents
Application of composition of strawberries with centella asiatica Download PDFInfo
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- CN111803546A CN111803546A CN202010552471.9A CN202010552471A CN111803546A CN 111803546 A CN111803546 A CN 111803546A CN 202010552471 A CN202010552471 A CN 202010552471A CN 111803546 A CN111803546 A CN 111803546A
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- composition
- centella asiatica
- strawberries
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- strawberry
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Classifications
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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)
- A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- 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)
- A61K36/23—Apiaceae or Umbelliferae (Carrot family), e.g. dill, chervil, coriander or cumin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
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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
- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P27/00—Drugs for disorders of the senses
- A61P27/16—Otologicals
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/06—Antihyperlipidemics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Diabetes (AREA)
- Mycology (AREA)
- Botany (AREA)
- Hematology (AREA)
- Epidemiology (AREA)
- Neurology (AREA)
- Neurosurgery (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Medical Informatics (AREA)
- Biotechnology (AREA)
- Obesity (AREA)
- Alternative & Traditional Medicine (AREA)
- Gastroenterology & Hepatology (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Nutrition Science (AREA)
- Endocrinology (AREA)
- Emergency Medicine (AREA)
- Psychiatry (AREA)
- Hospice & Palliative Care (AREA)
- Medicines Containing Plant Substances (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention belongs to the technical field of medicine preparation, and relates to a composition consisting of strawberries pubescens and centella asiatica. The composition of the strawberries and the centella asiatica prepared by the invention has the effects of reducing blood sugar and blood fat, improving the oxidation resistance of the liver and protecting the structure and the function of the liver, and can be used for improving hyperglycemia, weight loss and 'three more' symptoms caused by diabetes; can be used for reducing blood lipid level; can be used for improving learning and memory ability, protecting the connection between hippocampal nerve synapses and other symptoms, protecting hearing and repairing hearing damage, and has wide application prospect.
Description
The application is a divisional application of a patent application with the application number of 201710357076.3, which is filed on 19.5.2017 and the name of 'a composition of strawberries and centella asiatica and a preparation method and application thereof'.
Technical Field
The invention belongs to the technical field of medicine preparation, and relates to a composition of strawberries and centella asiatica, a preparation method thereof, and applications thereof in reducing blood sugar, reducing blood fat, improving oxidation resistance of organisms, protecting liver structures and functions and repairing hearing impairment.
Background
The strawberries huangmao are plants of the genus strawberry of the family rosaceae, are named as strawberries rustica and white strawberries, grow on hilly grasslands or under grove forests with the altitude of 700 + 3000m, are distributed in Yunnan, Guangxi, Sichuan, Guizhou and other places in China, have the effects of clearing away heat and toxic materials, promoting blood circulation and removing blood stasis, have better medicinal value and are rich in resources. According to the report of the world health organization, cardiovascular diseases are the first cause of death worldwide and seriously threaten the health of people. Cardiovascular diseases include diabetes, hyperlipidemia, and the like. Diabetes mellitus is a metabolic disease characterized by disorders of glucose metabolism and hyperglycemia caused by genetic or acquired defects in insulin secretion. Diabetes patients suffer from continuous rise of blood sugar, rapid weight loss, and abnormal functions of liver, kidney and pancreas, and in addition, the disordered metabolism of diabetes patients often generates a large amount of oxygen free radicals, causing other complications. Hyperlipidemia is a disease with abnormal blood lipid concentration caused by abnormal fat metabolism or abnormal operation, and a large amount of lipid deposited in the intima of an artery can narrow the lumen of the blood vessel, slow the blood flow and block the artery, thereby causing coronary heart disease and cerebrovascular disease.
Disclosure of Invention
The invention aims to provide a composition of strawberries and centella asiatica for reducing blood sugar and blood fat, improving the oxidation resistance of livers, protecting the structures and functions of the livers, enhancing the learning and memory ability and repairing hearing impairment.
The invention provides a composition of strawberries and centella, which comprises ethanol extracts of strawberries and centella; the composition comprises the following components in percentage by mass: 2-2: 1 leaching to obtain the product.
The invention also provides a preparation method of the composition of strawberries with strawberries, which comprises the following steps:
(1) drying whole plants of strawberries and centella asiatica, cutting, and extracting with ethanol to obtain extract;
(2) evaporating the extract prepared in the step (1), filtering and taking the supernatant to obtain the composition of the strawberries with the centella asiatica;
in the step (1), the concentration of the ethanol is 50% -95%; preferably, it is 75%.
In the step (1), the mass ratio of the strawberries with the centella is 1: 2-2: 1; preferably, 1: 1.
in the step (1), the volume ratio of the total mass of the strawberries and the centella asiatica to the ethanol is 12-30 g: 400-700 ml; preferably 24g to 500 ml.
In the step (1), the leaching temperature is 18-25 ℃; preferably, it is 25 ℃.
In the step (1), the leaching time is 2-4 hours; preferably, it is 3 hours.
In the step (1), the strawberries and centella asiatica comprise roots, stems and leaves of the plants.
In the step (2), the evaporation can be performed by a rotary evaporator.
In the step (2), the evaporation time is 1-2 hours; preferably, it is 1.5 hours.
In the step (2), the evaporation temperature is 35-45 ℃; preferably, it is 40 ℃.
In the step (2), the crude drug concentration of the composition of the strawberries with the centella asiatica is 0.8g/ml-1(ii) a Wherein the crude drug concentration refers to the concentration of the mixture stock solution obtained by preparation.
The invention also provides a composition of the strawberries pubescens and the centella asiatica, which comprises ethanol extracts of the strawberries pubescens and the centella asiatica.
The invention also provides a composition of the strawberries with the herba centellae, which is prepared by the preparation method and comprises ethanol extracts of the strawberries with the herba centellae.
The invention also provides application of the composition of the strawberries pubescens and the centella asiatica in preparing foods/health care products and medicines for reducing blood sugar.
The invention also provides application of the composition of strawberries pubescens and centella asiatica in preparing food/health care products and medicines for improving emaciation caused by diabetes.
The invention also provides application of the composition of the strawberries pubescens and the centella asiatica in preparing food/health care products and medicines for improving 'Sanduo' symptoms caused by diabetes.
The invention also provides application of the composition of the strawberries pubescens and the centella asiatica in preparing foods/health care products and medicines for reducing blood fat.
The invention also provides application of the composition of the strawberries pubescens and the centella asiatica in preparing food/health care products and medicines for improving the oxidation resistance of livers.
The invention also provides application of the composition of the strawberries pubescens and the centella asiatica in preparing food/health care products and medicines for protecting the structural function of the liver, in particular to protecting the structure and the function of the liver of a patient with hyperlipidemia.
The invention also provides application of the composition of the strawberries with the centella to preparation of food/health care products and medicines for enhancing learning and memory abilities.
The invention also provides application of the composition of strawberries pubescens and centella asiatica in preparing food/health care products and medicines for protecting normal connection between hippocampal neurons.
The invention also provides application of the composition of the strawberries pubescens and the centella asiatica in preparing foods/health care products and medicines for protecting hearing and repairing hearing damage.
The present study shows that the strawberry and centella asiatica composition of the present invention has the above significant beneficial effects, but the strawberry alone or the centella asiatica alone does not have the above effects.
The invention has the beneficial effects that the composition of the strawberries and the centella asiatica is used for treating the blood sugar reducing effect of STZ-induced diabetic mice and the blood fat reducing effect of hyperlipaemia rats for the first time, wherein the preparation method of the composition is simple to operate. The composition prepared by the method has obvious effects of reducing blood sugar rise and blood fat rise, can obviously improve the anti-oxidation level of the liver, and reduces the damage of oxygen free radicals to organisms. The strawberries composition prepared by the method can obviously improve the condition of liver tissue damage, protect the structure and the function of the liver, protect the structure and the function of hippocampal neurons and enhance the learning and memory ability. The composition prepared by the method can obviously recover the hearing damage of mice.
Drawings
FIG. 1A is the effect of the strawberry and centella asiatica composition on food intake in diabetic mice of example 1 and B is the effect of the composition on water intake in diabetic mice.
Fig. 2 is a graph of the effect of the strawberry and centella asiatica composition of example 1 on the oral glucose tolerance of diabetic mice (n-10).
FIG. 3A is the effect of the composition of strawberry and centella asiatica on food intake in the hyperlipidemic rats of example 1 and B is the effect of the composition on water intake in the hyperlipidemic rats.
FIG. 4 is the effect of the combination of strawberry and centella asiatica on the liver tissue structure of rats of example 1 (HE stained pathological section); a is a normal group, B is a high fat model group, C is a simvastatin group, and D is a high dose composition group.
FIG. 5 is a graph of the effect of the combination of strawberry (Physalis pubescens) and centella asiatica on the number and structure of liver mitochondria, endoplasmic reticulum (EBEM results of liver); a is a normal group, B is a high fat model group, C is a simvastatin group, and D is a high dose composition group.
FIG. 6 is a graph of the effect of the combination of strawberry and centella asiatica of example 1 on learning and memory in diabetic mice, where A is the escape latency and B is the percent residence time in the target quadrant.
FIG. 7 is a graph of the effect of the combination of strawberry and centella asiatica on hippocampal neurons of example 1 (hippocampal tissue electron microscopy); a is the normal group, B is the model group, C is the high dose group, and D is the metformin group.
Detailed Description
The present invention will be described in further detail with reference to the following specific examples and the accompanying drawings. The procedures, conditions, experimental methods and the like for carrying out the present invention are general knowledge and common general knowledge in the art except for the contents specifically mentioned below, and the present invention is not particularly limited.
Example 1 preparation of a composition of strawberries and centella asiatica
Drying herba Adenophorae Trachelioidis and herba Centellae whole plant (including root, stem, and leaf), cutting into pieces, weighing 12g each, extracting with 75% ethanol 500ml at room temperature for 3 hr, transferring to rotary evaporator, vacuum steaming, filtering to obtain supernatant, and making into crude drug with concentration of 0.8g/ml-1The composition of strawberries having a yellow hair and centella asiatica.
Example 2 STZ-induced diabetic mouse (STZ mouse) model construction
One week after the 120 mice were acclimatized, 10 mice were randomly selected as a normal group (control group). The rest mice induce type I diabetes model with STZ according to 40mg kg-1The dosage is injected into the abdominal cavity with 0.4 percent of STZ-sodium citrate solution. Normal groups were injected with equal amounts of distilled water intraperitoneally. Modeling for 1 week, fasting the mice for 6h, measuring fasting blood glucose value by glucometer, and discarding fasting blood glucose lower than 15 mmol.L-1Or more than 30 mmol.L-1Selecting mice with fasting blood glucose higher than 15 mmol.L-1And less than 30 mmol. L-1The mice were used as STZ-induced diabetic mice. Mice that were successfully modeled and had close and no significant differences in blood glucose values were equally divided into model group, metformin group, low dose group, high dose group, and strawberry group.
Example 3 modeling of hyperlipidemic rat
After 70 SD rats are adaptively fed for 1 week, 10 rats are randomly selected as a normal group of blank control, common feed is fed, the rest 60 rats are fed with high-fat feed to establish a hyperlipemia rat model, all rats freely drink water, after 20 days, the rats are fasted and are not forbidden to drink water for 12 hours, and tail vein blood is taken to measure four blood lipid indexes of TC, TG and LDL-C, HDL-C in serum. From the groups, 50 hyperlipidemic rats with better modeling effect (the blood lipid level is obviously higher than that of the normal group) and close body mass and blood lipid indexes without significant difference are obtained and averagely divided into 5 groups, namely a high-fat model group, a simvastatin group, a low-dose group of a Huangmao strawberry composition, a high-dose group and a Huangmao strawberry group.
Example 4 Effect of Fragaria Nigria and centella asiatica composition on fasting plasma glucose in STZ mice
Each group of mice in example 2 was treated at a dose of 10 mL/kg-1Volume lavage, lavage of distilled water in normal group (control group) and model group, and lavage of 0.4 g.kg in low dose group and high dose group respectively-1And 0.8 g.kg-1The composition of strawberry and centella asiatica prepared in example 1, metformin (positive group) and gavage 0.15g/kg-1Metformin, xanthophyllum strawberry group intragastric administration 0.4 g.kg-1The strawberry extract is extracted for 1 time/d for 4 weeks. The blood sugar of the mice is measured every week, and the blood is collected from the tail vein after fasting for 6h, and the fasting blood sugar is measured.
The experimental results are shown in table 1, and the composition of strawberries pubescens and centella asiatica prepared in example 1 can significantly reduce the fasting blood glucose level of diabetic mice, and the higher the concentration is, the more obvious the effect is. The result shows that the composition of the strawberries and the centella asiatica has the function of reducing the fasting blood glucose.
TABLE 1 EXAMPLE 1 Effect of Fragaria Nigria and centella asiatica compositions on fasting blood glucose levels in STZ mice ((
n＝10)
Note: in comparison to the set of models,*P＜0.05，**P＜0.01
example 5 example 1 Effect of strawberry and centella asiatica composition on body weight of STZ mice
The body weight of the mice was measured by gavage as in example 4.
Table 2 example 1 effect of strawberry and centella asiatica composition on body weight of STZ mice ((
n＝10)
Note: in comparison to the set of models,*P＜0.05，**P＜0.01
example 6 example 1 Effect of Fragaria maackii and centella asiatica composition on food intake and water intake in STZ mice
The mice were gavaged as described in example 4 and their water intake and food intake were measured daily.
As shown in FIG. 1, the composition of strawberry and centella asiatica of example 1 is effective in improving the weight loss of diabetic mice and significantly improving the intake of water and food. The results show that the strawberries composition has the effects of increasing the body weight of diabetic mice and improving the polydipsia and polyphagia of the diabetic mice.
Example 7 example 1 Effect of a combination of Fragaria Nibea and centella asiatica on the OGTT of STZ mice
Oral Glucose Tolerance Test (OGTT) was performed 1d after the last administration (example 4 after 4 weeks of gastric gavage). The mice are fasted for 5h, and after eating for 1h normally, the mice are fed with 2 g/kg-glucose solution for intragastric administration, and the blood sugar values are measured at 0, 30, 60 and 120min after glucose treatment respectively.
The experimental result is shown in fig. 2, after 30min of glucose treatment, the blood sugar of the model group mice is continuously increased, the composition of the strawberries and the centella asiatica can effectively recover the blood sugar value of the mice at 60min, and the blood sugar value is close to the self level at 120 min. The experimental result shows that the composition of the strawberries and the centella asiatica in example 1 can effectively repair and improve the glucose tolerance of diabetic mice, and the result shows that the composition of the strawberries and the centella asiatica has an obvious blood sugar reducing effect.
Example 8 example 1 Effect of Fragaria Nigri and centella asiatica compositions on blood lipid and antioxidant capacity in STZ mice
After the completion of the OGTT test in example 7, blood was collected from the mouse eyeballs, and liver tissue was immediately removed after sacrifice to prepare liver homogenate. Standing whole blood in ice bath for 15min, centrifuging at 2000rpm for 20min, and determining superoxide dismutase (SOD), Catalase (CAT) activity and Malondialdehyde (MDA) content in liver, all according to the kit specification.
The experimental results are shown in table 3, compared with the normal group (control group), the contents of SOD and CAT in the model group are obviously reduced, and the MDA accumulation is obviously increased, which shows that the liver antioxidant level of the model group is obviously reduced, and the composition of strawberries and centella asiatica in example 1 can effectively recover the blood lipid level of a diabetic mouse, and can obviously improve the liver antioxidant capacity of the diabetic mouse.
TABLE 3 Effect of Fragaria majus and centella asiatica composition on blood lipid and liver antioxidant levels in STZ mice ((
n＝10)
Note: p < 0.05, P < 0.01, compared to model group
Example 9 example 1 Effect of a combination of Fragaria Nigria and centella asiatica on the body weight of hyperlipidemic rats
During the experimental gavage period, rats in the normal group (control group) were fed with normal diet, and the model group, simvastatin group, and low dose group, high dose group, and strawberry group were continuously fed with high fat diet. Each group was measured at 10 mL/kg-1Performing intragastric administration by volume, wherein the normal group and the model group are intragastric administered with distilled water, and the simvastatin group is administered with intragastric administration of 0.4mg & ml-1The simvastatin solution of (1) is used for intragastric administration of 0.4 g/ml for the low dose group and the high dose group respectively-1、0.8g·ml-1Yellow of (2)Composition of strawberry and herba Centellae, and gavage of strawberry group of 0.4 g/ml-1The amount of the strawberry extract is 1 time/d, and the total period is 3 weeks. Body weight was measured once per week during the gavage period.
As shown in table 4, the body weight of the rats in the model group was significantly increased compared to that in the normal group (control group); experimental results show that the composition of the strawberries mallow and the centella asiatica in the example 1 can effectively improve the weight aggravation condition of diabetic rats, and the higher the concentration of the composition is, the more obvious the effect is, which shows that the composition can effectively reduce the weight of hyperlipidemia.
Table 4 example 1 effect of a combination of strawberries and centella asiatica on the body weight of hyperlipidemic rats (n ═ 10)
Note: p < 0.05, P < 0.01 compared to model group.
Example 10 example 1 Effect of the combination of Fragaria maackii and centella asiatica on intake and drinking of hyperlipidemic rats
Experimental procedures As in example 9, rats were daily recorded for water and food intake during the gavage period.
The results are shown in FIG. 3, where A is the effect of the strawberry and centella asiatica composition of example 1 on the intake of rats and B is the effect of the composition on the intake of rats. The rats in the model group had significantly increased food and water intake compared to the normal group (control group); the experimental result shows that the composition can obviously reduce the intake and drinking amount of the hyperlipoidemia rats, and the higher the dosage is, the more obvious the effect is.
Example 11 example 1 Effect of a combination of Fragaria maurensis and centella asiatica on blood lipids in hyperlipidemic rats
In example 9, after the administration for 3 weeks, fasting was performed without water prohibition for 12 hours, blood was taken from the tail vein, whole blood was allowed to stand in a refrigerator at 4 ℃ for 15 minutes, centrifuged at 3000rpm for 15 minutes, and the supernatant was aspirated to obtain serum. And four blood lipid indexes of TC, TG and LDL-C, HDL-C in serum are measured according to the operation of a kit instruction.
The experimental results are shown in table 5, and compared with the normal group (control group), the blood lipid content in the serum of the rat in the high-fat model group is obviously increased; experimental results show that the composition of the strawberries mallow and the centella asiatica in the example 1 can obviously reduce the content of TC, TG and LDL-C in serum and obviously improve the content of HDL-C, and the higher the concentration is, the more obvious the effect is.
TABLE 5 EXAMPLE 1 Effect of strawberry and centella asiatica composition on the blood lipids of hyperlipidemic rats (n ═ 10)
Note: p < 0.05, P < 0.01 compared to model group.
Example 12 example 1 Effect of a combination of Fragaria maurensis and centella asiatica on antioxidant levels in hyperlipidemic rats
In example 9, after the administration for 3 weeks, fasting was not performed for 12 hours, and after the mice were sacrificed, liver tissues were immediately taken out, and after homogenization, the superoxide dismutase (SOD) activity and Malondialdehyde (MDA) content in the liver were measured according to the kit instructions.
The experimental results are shown in table 6, compared with the normal group, the high-fat model group has significantly reduced SOD content, which indicates that the liver oxidation resistance level is significantly reduced; the experimental result shows that the composition of the strawberries and the centella asiatica in the example 1 obviously reduces the content of MDA, obviously improves the SOD activity, and has more obvious effect when the dosage is higher. The results demonstrate that the strawberry and centella asiatica composition of example 1 has a significant effect of improving the antioxidant capacity of the liver.
Table 6 example 1 effect of a combination of strawberries from huangmao and centella asiatica on MDA and SOD in the liver of hyperlipidemic rats (n ═ 10)
Note: p < 0.05, P < 0.01 compared to model group.
Example 13 example 1 Effect of Fragaria maackii and centella asiatica composition on hepatic tissue architecture, hepatocyte nuclei, mitochondria and endoplasmic reticulum in hyperlipidemic rats
In example 9, after completion of administration, the rats were sacrificed, and livers were immediately taken out and fixed with 10% formaldehyde, embedded in paraffin, sectioned, and stained with HE conventionally to prepare fixed sections, which were then sectioned by paraffin observation.
The results are shown in fig. 4, the liver tissue structure of the model group is atrophied, vacuolation is obvious, the stem cell morphology is shrunk, the liver tissue structure of the strawberry and centella asiatica composition group in example 1 is clear, hepatic cell necrosis is rare, lipid droplets in cytoplasm are less, and the results show that the composition has a protective effect on the liver tissue.
In example 9, after completion of administration, the rat was sacrificed, and the peripheral part of the liver lobe was immediately taken out, cut into 1mm × 1mm tissue pieces, fixed with 2.5% glutaraldehyde, kept at 4 ℃ in a refrigerator for use, and stained fixed sections were observed by transmission electron microscope.
The results are shown in fig. 5, the cell nuclei of the model group are fixedly contracted, the mitochondria are swollen, loose and vacuolated obviously, and the number of rough endoplasmic reticulum is reduced and the arrangement is loose; the high-dose composition group can ensure complete shape of nucleus cells, compact arrangement of mitochondria and obviously increased number of rough endoplasmic reticulum.
EXAMPLE 14 Effect of the composition on learning and memory in diabetic mice
After gavage as in example 4, the water maze was divided into four quadrants, labeled 1, 2, 3, and 4, and the platform was placed in quadrant 4. A circular platform with a diameter of 5cm was placed in the center of the first quadrant. The tank water just submerges the platform. Mice were randomly placed from one quadrant on the first day of the experiment, and pushed forward one quadrant each day thereafter. If the mouse finds the platform within 60S, recording the latency period, and making the mouse return to the mouse cage after the mouse stays on the platform for 10S; if the platform can not be found in 60S, the latency period is 60S, and meanwhile, the mouse cage needs to be returned after the platform is manually guided to stay for 10S. The experiment was repeated for the next start site until all 4 quadrants of the day were completed and training was continued for five consecutive days.
And after the directional navigation experiment is finished, removing the platform during the experiment. And (3) putting the mouse into the opposite quadrant of the platform, taking the mouse out after 60 seconds, putting the mouse into a mouse cage, and recording the residence time percentage of the target quadrant of the mouse and a mouse swimming route map.
As shown in fig. 6A, the escape latency time of the model group is increased, the latency of the mice can be significantly reduced by the composition, and the effect is more obvious at higher concentration. As shown in fig. 6B, the retention percentage of the target quadrant of the mice in the composition group is obviously increased and is close to that of the normal group (control group), and the experimental result shows that the composition can obviously improve the learning and memory abilities of the mice.
EXAMPLE 15 Effect of the composition on hippocampal neurons in STZ mice
Example 4 after completion of gavage, mice were sacrificed, and hippocampal tissue of the brain was immediately taken out, cut into tissue pieces of 1mm × 1mm, fixed with 2.5% glutaraldehyde, kept at 4 ℃ in a refrigerator for future use, and stained fixed sections were observed by transmission electron microscopy.
As shown in FIG. 7, in the model group, there was mitochondrial depletion in hippocampal neurons, decreased synapse numbers in neurons, enlarged synapses, swelling, increased gaps, larger neuron bodies in the composition group, significantly increased mitochondrial numbers, close connection with peripheral neurons, and significantly increased synapse numbers. The result shows that the composition can obviously improve the connection between the hippocampal neurons and has the effect of protecting the hippocampal neurons.
Example 16 construction of mouse model with hearing impairment
After 60 Kunming mice are bred adaptively for one week, 10 Kunming mice are randomly selected as a normal group (a control group), the other mice are exposed to white noise of 4-45 kHZ and 110dBSPL for 9h every day for 10d, auricle reflex, namely after the phenomenon that auricles are erected to adapt to sharp sounds is eliminated, the auricles are randomly divided into a model group, a flunarizine hydrochloride group, a Huangmao strawberry group, a low dose group and a high dose group.
Example 17 Effect of the combination of Fragaria Nigria and centella asiatica on the auditory ability of mice
Each group of mice was treated with 10 mL/kg-1Volume intragastric administration, intragastric administration of distilled water for normal group (control group) and model group, intragastric administration of flunarizine hydrochloride group of 1.5X10-3g·kg-1Intragastric administration of 0.4 g.kg in the low and high dose groups-1And 0.8 g.kg-1Yellow wool prepared in example 1Strawberry and centella asiatica composition, wherein the weight of the strawberry group is 0.4 g/kg-1An extract of Fragaria Nipponica. 1/d for 4 weeks. After the completion of the gavage, cochlear electrograms (ECochG) and Auditory Brainstem Response (ABR) were measured in each group of mice.
Mice are anesthetized by intraperitoneal injection of 4mg/kg phenobarbital sodium solution, and ABR and ECochG experiments are carried out by adopting a YSSD-1000 multifunctional evoked potential instrument, wherein the stimulation sound is short wave, the sound pressure level is 90dB, the sensitivity is 6.2uv, and the band-pass frequency is 100-1000 HZ. The mouse binaural individual wave latency, wave interval and Compound Action Potential (CAP) amplitude changes were recorded.
Effect on CAP amplitude as shown in table 7, CAP amplitude was significantly increased in the case of the strawberries and centella asiatica composition (low dose group and high dose group) compared to the model group. The results show that the composition of strawberries pubescens and centella asiatica prepared in example 1 can remarkably recover the hearing impairment of mice, and the effect is more obvious when the concentration is higher.
Effect on latency and wave interval results as shown in table 8, I, III wave latencies and I-IV and I-V wave intervals of the strawberry and centella asiatica composition group (low dose group and high dose group) were significantly restored to the strawberry and centella asiatica composition prepared in example 1, which significantly improved auditory central function, compared to the model group. The result shows that the composition of the strawberries pubescens and the centella asiatica has the effects of treating and repairing the noise hearing impairment.
Table 7 example 1 effect of strawberry and centella asiatica composition on CAP amplitude ((s))
n＝10)
Note: p < 0.05, P < 0.01, compared to model group
Table 8 example 1 effect of strawberry and centella asiatica compositions on latency and wave interval: (
n＝10)
Note: p < 0.05, P < 0.01, compared to model group
The protection of the present invention is not limited to the above embodiments. Variations and advantages that may occur to those skilled in the art may be incorporated into the invention without departing from the spirit and scope of the inventive concept, and the scope of the appended claims is intended to be protected.
Claims (10)
1. Application of a composition of strawberries pubescens and centella asiatica in preparing foods/health products and medicines for protecting hearing and repairing hearing damage.
2. Application of composition of herba Fragariae Nigri and herba Centellae in preparing food/health product and medicine for enhancing learning and memory ability and protecting normal connection between hippocampal neurons is provided.
3. The use according to claim 1 or 2, wherein the strawberry and centella asiatica composition comprises an ethanol extract of strawberry and centella asiatica.
4. Use according to claim 1 or 2, wherein the strawberry and centella asiatica composition is prepared by:
(1) drying and chopping strawberries and centella asiatica, and extracting with ethanol to prepare extract;
(2) evaporating the extract prepared in the step (1), filtering and taking the supernatant to obtain the composition of the strawberries and the centella asiatica.
5. The use of claim 4, wherein in step (1), the ethanol concentration is 50% to 95%.
6. The use according to claim 4, wherein in the step (1), the mass ratio of the strawberries and the centella asiatica is 1: 2-2: 1.
7. the use according to claim 4, wherein in step (1), the volume ratio of the total mass of the strawberries and centella asiatica to the ethanol is 12-30 g: 400 and 700 ml.
8. The use according to claim 4, wherein in step (1), the temperature of the leaching is 18-25 ℃; the leaching time is 2-4 hours.
9. The use according to claim 4, wherein in step (2), the temperature of the evaporation is 35 ℃ to 45 ℃; the evaporation time is 1-2 hours.
10. Use according to claim 1 or 2, characterized in that said composition of strawberry and centella asiatica is prepared by a process comprising: drying whole plant of herba Fimbristylis Dichotomae and herba Centellae, cutting into pieces, weighing 12g each, soaking and extracting with 75% ethanol 500ml at room temperature for 3 hr, transferring to rotary evaporator, vacuum steaming, filtering to obtain supernatant, and making into composition of herba Fimbristylis Dichotomae and herba Centellae with crude drug concentration of 0.8g ml-1.
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