CN111686139B - Application of pine nut shell water extract in blood lipid reducing medicine - Google Patents

Application of pine nut shell water extract in blood lipid reducing medicine Download PDF

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CN111686139B
CN111686139B CN202010638000.XA CN202010638000A CN111686139B CN 111686139 B CN111686139 B CN 111686139B CN 202010638000 A CN202010638000 A CN 202010638000A CN 111686139 B CN111686139 B CN 111686139B
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pine nut
nut shell
pine
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fat
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CN111686139A (en
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领小
鲍明明
苏雅拉图
孟和
昭日格图
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Inner Mongolia University
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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
    • A61K36/13—Coniferophyta (gymnosperms)
    • A61K36/15—Pinaceae (Pine family), e.g. pine or cedar
    • 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
    • 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
    • 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
    • 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
    • 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
    • A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
    • 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
    • A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
    • 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
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
    • 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
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization

Abstract

The invention provides an application of a Korean pine nut shell water extract in preparation of a blood fat reducing medicine or a blood fat reducing health care product, belonging to the technical field of medicines. Pulverizing pine nut shell of Korean pine, adding water, refluxing, and concentrating the filtrate to obtain water extract of pine nut shell. The administration of the pine nut shell aqueous extract to the high-fat model rat by gastric lavage can remarkably reduce the serum Total Cholesterol (TC), Triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C) of the high-fat model rat and remarkably increase the serum high-density lipoprotein cholesterol (HDL-C) of the high-fat model rat. The pine nut shell water extract of the Korean pine has the function of regulating blood fat, and the pine nut shell is the waste of the pine nut of the Korean pine, has easily obtained, cheap and simple raw materials, and can be applied to the preparation of medicaments for preventing and treating hyperlipidemia and health-care food for reducing blood fat.

Description

Application of pine nut shell water extract in blood lipid reducing medicine
Technical Field
The application relates to the technical field of medicines, in particular to application of a Korean pine nut shell water extract in preparation of a lipid-lowering medicine.
Background
Blood lipids are a general term for neutral fats and lipids in plasma, and the main components include Triglyceride (TG), Total Cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and the like. The increase of one or more lipid levels in blood (decrease of HDL-C) above normal level is called hyperlipidemia (hyperlipidemia), abbreviated as hyperlipidemia. Dyslipidemia is generally not expressed abnormally in the body, but persistent hyperlipidemia, lipid in blood is deposited on the arterial wall to form atherosclerotic plaque, cardiovascular and cerebrovascular diseases such as hypertension, myocardial anoxia, myocardial infarction, cerebral infarction and the like are induced, and hyperlipidemia is a potential invisible killer in the body.
Currently, lipid-lowering drugs in clinical use are mainly: statins, fibrates, nicotinic acids, bile acid sequestrants, unsaturated fatty acids, and the like. However, these drugs have various degrees of side effects such as gastrointestinal reaction, nausea, constipation or diarrhea, headache, dizziness, liver injury, etc. China has abundant natural resources and a long history of natural medicines (such as traditional Chinese medicines). The natural medicine has the advantages of definite curative effect, high safety, low toxicity, low price and the like and is more and more favored by people.
Pine nut, a seed of Korean pine, is both a traditional Chinese medicine and a food. Pine nut as Chinese medicine has the functions of invigorating kidney, benefiting vital energy, nourishing blood, moistening intestine, relaxing bowels, moistening lung, etc. The product is a longevity fruit, which can be eaten for a long time to strengthen body and mind, moisten skin and prolong life. China is the production of pine nuts and imports and exports of big countries, and the annual production of pine nuts of Korean pine is about 1 ten thousand tons. The pine nut shells and pinecones, which are by-products formed during the pine nut processing, account for about 70% of the entire pine cone. Due to insufficient attention, the wastes are mostly only used as firewood except for a small amount of dye and active carbon, which causes great waste of resources.
The inventor finds that the red pine nut shell water extract has the effects of preventing and treating hyperlipidemia of a high-fat model rat in pharmacological research of the red pine nut shell.
In view of this, the present application is specifically made.
Disclosure of Invention
The invention aims to provide application of a Korean pine nut shell water extract in preparation of a lipid-lowering drug. The pine nut shell water extract for treating hyperlipidemia is simple and easily obtained in raw materials, low in cost and less in toxic and side effects.
In a first aspect, the present application provides an aqueous extract of pine nut shells of Korean pine, which is prepared from pulverized pine nut shell powder of Korean pine, an appropriate amount of distilled water, by heating under reflux, filtering, concentrating, and drying.
The raw materials are simple and easy to obtain, and can be used for regulating fat, protecting liver and improving the disease resistance of the whole body.
In a second aspect, the embodiments of the present application provide a use of an aqueous extract of pine nut shells of Korean pine in a lipid-lowering drug.
Compared with the prior art, the invention has the beneficial effects and characteristics that:
(1) not only reducing the content of total cholesterol in serum; it also can reduce triglyceride and low density lipoprotein cholesterol; and simultaneously, the content of high-density lipoprotein cholesterol is increased.
(2) Compared with chemical lipid-lowering medicines and health-care products, the pine nut shells belong to the waste materials of pine nuts, and have the advantages of easily available sources, low price and less adverse reactions under the condition of equivalent medicine effect.
(3) Although the pine nut shells of the Korean pine are wastes, the pine nut shells of the Korean pine contain active ingredients such as volatile oil, polyphenols, flavonoids, procyanidine, polysaccharide and the like, and the pine nut shells have various biological activities such as enhancing the body immunity, resisting tumors, resisting oxidation, resisting bacteria, resisting viruses and the like according to literature reports. The invention provides the application of the pine nut shells of the Korean pine in the preparation of the hypolipidemic drugs.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below.
FIG. 1 shows the results of measurement of the effect of pine nut shells of Korean pine on the Total Cholesterol (TC) content in serum of rats.
FIG. 2 is a graph showing the results of measurement of the effect of pine nut shells of Korean pine on the serum Triglyceride (TG) content in rats.
FIG. 3 shows the results of measurement of the effect of pine nut shells of Korean pine on the serum low-density lipoprotein cholesterol (LDL) content in rats.
FIG. 4 is a graph showing the results of measurement of the effect of pine nut shells of Korean pine on the serum high-density lipoprotein cholesterol (HDL) content of rats.
Very significant differences (P < 0.01) compared to model group; significant differences (P < 0.05).
Detailed Description
1. Blood fat reducing drug effect experiment of Korean pine nut shell water extract
(1) Experimental Material
Experimental drugs and reagents:
pine nut shells of Korean pine, purchased from Jilin Changchun.
Simvastatin, manufactured by guangdong petidi pharmaceutical co ltd; approval document No.: standard Chinese character 20066221
A total cholesterol kit produced by Zhongsheng Bei accuse Biotechnology GmbH; product batch number: 141741, product registration number: jing Yao Zhen machinery (Standard) 2011 No. 2400202
A triglyceride kit produced by Zhongsheng north controlled Biotechnology GmbH; production batch number: 136331, product registration number: jing Yao Zhen machinery (Standard) 2011 No. 2400202
The high-density lipoprotein cholesterol kit is produced by Zhongsheng Bei Zhi Biotech GmbH; production batch number: 140551, product registration number: jing Yao Zhen machinery (Standard) 2011 No. 2400202
The low-density lipoprotein cholesterol kit is produced by Zhongsheng Bei Zhi Biotechnology GmbH; production batch number: 140571, product registration number: jing Yao Zhen machinery (Standard) 2011 No. 2400202
The cholesterol is produced by Beijing double-rotation microbial culture medium product factories; production batch number: 20140621
The sodium cholate is produced by a microorganism culture medium product factory in the Haihu district of Beijing; production batch number: 0140516
An experimental instrument:
model BJ-200 high-speed multi-function pulverizer (deqing byjie electric appliances ltd); DT-500 electronic balance (Jiangsu double-jie test instruments factory, everlasting maturity); DGSY-III electric heating constant temperature water bath (Beijing, Italy, science and technology development company); RE-3000A rotary evaporator (Shanghai Yangrong Biochemical Instrument factory); TDL-5 centrifuge (Shanghai' an pavilion scientific instruments factory); PRONTO EVOLUTION full-automatic biochemical instrument (Italy).
Experimental animals and feed:
wistar rats 40 (provided by the Experimental animal center of university of inner Mongolia, production license number: SCXK (Mongolia) 2016-0001) weighed 180. + -.20 g.
High-fat feed: 0.5 percent of sodium cholate, 3 percent of cholesterol, 10 percent of lard oil and 86.5 percent of basic feed (flour 20 percent, rice flour 10 percent, corn 20 percent, bran 25 percent, bean material 20 percent, bone meal 2 percent, fish meal 2 percent, salt 0.9 percent and vitamin 0.1 percent).
(2) Experimental methods
Preparation of aqueous extract of pine nut shell of Korean pine
Cleaning pine nut shells, crushing, and screening with a 18-mesh screen. 50 g of the powder was weighed, added with 300 mL of distilled water, heated under reflux for 2 h, and filtered. Adding 200 mL of water into filter residue, heating and refluxing for 1.5 h, and filtering. Adding 200 mL of water into the filter residue for the third time, heating and refluxing for 1.5 h, and filtering. The filtrates were combined, concentrated by rotary evaporator, and freeze-dried to give 1.21 g of extract with an extraction rate of 2.43%.
Feeding and administration modes:
wistar male rats are bred in an environment-adaptive mode for 7 days, fed with basic feed and freely drunk with water, the temperature is 23 +/-2 ℃, the humidity is 40% -50%, artificial illumination is carried out, and the light and the shade of the animal are 12 hours in each animal room. Rats were randomly divided into four groups, a blank group, a high-fat model group, a positive group, a pine nut shell group, and the like, with 10 rats per group. The positive group was gavaged with 10 mg/kg simvastatin, the pine nut shell group was gavaged with 30 mg/kg pine nut shell water extract, and the high fat model group and the blank group were gavaged with distilled water of the same volume. The gavage is carried out once a day for 14 days continuously. The blank group was fed with basal diet, and the other groups were fed with high fat diet.
(3) Observation indicator and measurement method
During the experiment, the body weight was weighed 1 time per day and the appetite, behavior, feces, hair and animal death were observed. After 14 days of gastric lavage, the patient is fasted for 16 hours and freely drinks water. Blood was taken from the femoral artery, stood for 30 min, centrifuged at 3000 r/min for 15 min, and serum was separated. The full-automatic biochemical analyzer adopts an enzyme colorimetric method to measure the Total Cholesterol (TC) value and the Triglyceride (TG) value of serum. Phosphotungstic acid-magnesium precipitation method for determining high density lipoprotein cholesterol (HDL-C) value and polyethylene sulfuric acid precipitation method for determining low density lipoprotein cholesterol (LDL-C) value.
The data obtained are statistically processed by means of the mean. + -. standard deviation (
Figure 542874DEST_PATH_IMAGE001
±s) Showing the significance of the differencetAnd (6) checking and judging.PThe calculation method of the value is as follows: the group of high-fat models was compared with the blank group, and each administration group was compared with the group of high-fat models. (P)<0.01 is a very significant difference, P<0.05 is a significant difference).
(4) Blood fat reducing experimental result and comparative analysis
During the experiment, the rats in each group have good mental status, normal weight increase, normal hair luster and activity, and no abnormal conditions such as death of the rats and the like.
The blood lipid indices of each group are shown in Table 1 and FIGS. 1-4, respectively.
TC of blank group: 1.54. + -. 0.36 mmol/L, TG: 0.55. + -. 0.19 mmol/L, LDL-C: 0.88. + -. 0.14 mmol/L, HDL-C: 0.63 plus or minus 0.15 mmol/L, and the blood lipid values of the above components in the high lipid model group are respectively 6.31 plus or minus 0.53 mmol/L, 0.87 plus or minus 0.14 mmol/L, 0.23 plus or minus 0.11 mmol/L and 2.92 plus or minus 0.58 mmol/L. The TC, TG and LDL-C values of the serum of the rat in the high-fat model group are increased compared with those in the blank group, and have extremely obvious difference (P < 0.01); the HDL-C value is reduced, and the HDL-C value is very different (P < 0.01), so that the high fat model is proved to be successful in modeling.
The positive group and the pine nut shell water extract group have extremely obvious difference (P < 0.01) in comparison with the high fat model group in the reduction of TC.
Compared with the high-fat model group, the positive group and the pine nut shell water extract group have very significant difference in TG reduction (P < 0.01).
The positive group, the pine nut shell water extract group, had a decreased LDL value compared to the high fat model group, but not significantly. The positive group, the pine nut shell water extract group and the high fat model group increase HDL-C with significant difference (P < 0.05).
TABLE 1 rat blood normative values on day 14 of administration of pine nut shell aqueous extract (n=10 unit mmol/L)
Group of TC TG HDL-C LDL-C
Blank group 1.54±0.36** 0.55±0.19** 0.63±0.15** 0.88±0.14**
Model set 6.31±0.53 0.87±0.14 0.23±0.11 2.92±0.58
Positive group 3.67±0.74** 0.28±0.14** 0.40±0.23* 2.30±0.25
Pine nut shell group 4.15±0.62** 0.37±0.13** 0.41±0.18* 2.65±0.35
Note: comparison with model groups: p <0.01 very significant difference; p <0.05, significant difference.
(5) Results and conclusions of the experiment
The Korean pine nut shell water extract can remarkably reduce TC and TG in serum of a rat with a high-fat model; the pine nut shell water extract of the Korean pine can reduce LDL in serum of rats in a high-fat model; the water extract of the pine nut shell of the Korean pine can obviously improve the HDL-C of the serum of a rat. In a word, the pine nut shell water extract of the Korean pine has the function of regulating blood fat and is a potential blood fat reducing medicine.
2. Acute toxicity test of Korean pine nut shell water extract
(1) Experimental materials and animals
Pine nut shells of Korean pine, purchased from Jilin Changchun.
20 Wistar rats (provided by the experimental animal center of university of inner Mongolia and having production license number: SCXK 2016-.
An experimental instrument:
model BJ-200 high-speed multi-function pulverizer (deqing byjie electric appliances ltd); DT-500 electronic balance (Jiangsu double-jie test instruments factory, everlasting maturity); DGSY-III electric heating constant temperature water bath (Beijing, Italy, science and technology development company); RE-3000A rotary evaporator (Shanghai Yangrong Biochemical Instrument factory); TDL-5 centrifuge (Shanghai' an pavilion scientific instruments factory).
(2) Experimental methods
Preparation of aqueous extract of pine nut shell of Korean pine
The preparation method is the same as example 1.
Feeding and administration mode
Taking 20 Wistar rats with half male and half female, weighing 180 +/-20 g, randomly dividing into an administration group and a blank group, 10 rats with half male and half female, and feeding basic feed. During the acute toxicity test, the maximum dose is fed once. The dosage of the pine nut shell water extract of the Korean pine is 0.6 g/kg body weight, and the blank group is filled with distilled water with the same volume as the stomach. The general reaction and the toxic condition of the rats are observed, 14 days are continuously observed, and the feeding condition is ensured to be unchanged during the observation period.
(3) Results and analysis of the experiments
Rat gavage0.6 g/kg of pine nut shell water extract of the Korean pine is administrated, and the dosage is 20 times of the dosage of a pharmacodynamic experiment. The stomach was gavaged once and observed continuously for 14 d. No obvious toxic side effect and death condition of rats appear in the experimental process, and LD can not be detected50，LD50>0.6 g/kg. The general toxic reaction of the pine nut shell water extract of the Korean pine is small, and the general toxic reaction of the pine nut shell water extract of the Korean pine basically has no toxic reaction to wistar rats.
The above-described embodiments are merely preferred embodiments of the present invention, rather than all embodiments, and are not intended to limit the present invention, which may be modified and varied by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (5)

1. An application of the aqueous extract of pine nut shell in preparing the medicines for decreasing blood fat is prepared from the powdered pine nut shell of red pine through pulverizing, adding distilled water, heating, reflux, filtering, concentrating and drying.
2. Use according to claim 1, wherein the aqueous extract of pine nut shells is used for the preparation of a medicament for reducing total cholesterol in serum.
3. The use of claim 1, wherein the aqueous extract of pine nut shells is used in the preparation of a serum triglyceride lowering medicament.
4. The use of claim 1, wherein the aqueous extract of pine nut shells is used for the preparation of a medicament for lowering low density lipoprotein cholesterol in blood serum.
5. The use of claim 1, wherein said aqueous extract of pine nut shells is used in the manufacture of a medicament for the prevention of a decrease in serum HDL cholesterol.
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