CN114073717A - Application of inonotus obliquus extract in reducing homocysteine - Google Patents
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Abstract
The invention discloses an application of inonotus obliquus extract in preparing a medicine and/or a functional food for treating or preventing hyperhomocysteinemia, and provides a rat experimental model of hyperhomocysteinemia induced by feeding methionine feed through gastric perfusion of the inonotus obliquus extract, so that the inonotus obliquus extract has the effect of obviously reducing the concentration of homocysteine in serum, and the application prospect of the inonotus obliquus extract in the aspect of preventing and treating hyperhomocysteinemia is shown.
Description
Technical Field
The invention belongs to the technical field of medicines, and relates to a medical application of an inonotus obliquus extract in reducing homocysteine.
Background
Hyperlipidemia is a major complication of cardiovascular and cerebrovascular diseases, diabetes, hypertension and the like, and seriously threatens human health. The current blood fat reducing drugs mainly comprise statins, cholesterol absorption inhibitor drugs and fibrates, and the action mechanism of the drugs mainly achieves the effect of reducing blood fat by inhibiting the absorption and synthesis of cholesterol. Lipid-lowering chemical drugs have great side effects, such as liver function damage, muscle damage, increased diabetes risk and the like.
Inonotus obliquus (Inonotus obliquus) is a fungus used as both medicine and food, and is also called Fuscoporia obliqua. The sclerotium of Inonotus obliquus is a tuber-shaped sterile fruiting body, has a yellow brown to black surface, is hard and uneven, and is yellow after being crushed. The inonotus obliquus is mainly parasitized under the bark of white birch, silver birch, elm, alnus japonica and the like, and the growing area of the inonotus obliquus is mainly distributed in a cold area with north latitude of 40-50 ℃ in northern hemisphere.
The active components of Inonotus obliquus mainly comprise polysaccharide and triterpene. At present, in-vivo and in-vitro experimental researches show that the inonotus obliquus has various pharmacological activities, including tumor resistance, oxidation resistance, blood sugar reduction, blood fat reduction, inflammation resistance, immune regulation and the like.
Research shows that the inonotus obliquus extract and various bioactive substances can inhibit the growth of various cancer cells, and the mechanism of the inonotus obliquus extract and the bioactive substances can be related to cell cycle regulation, immunoregulation and antioxidation.
Experiments of various diabetes animal models show that the inonotus obliquus crude polysaccharide or water extract shows obvious blood sugar reducing effect, and can reduce total Triglyceride (TG), Total Cholesterol (TC) and low density lipoprotein (LDL-C) in serum of the diabetes animal models, and the action mechanism of the inonotus obliquus crude polysaccharide or water extract is possibly related to inhibition of inflammatory reaction, promotion of sugar absorption, increase of insulin sensitivity, antioxidation and the like.
Experiments of high-fat diet-induced hyperlipemia rats show that the inonotus obliquus polysaccharide can obviously reduce serum total Triglyceride (TG), Total Cholesterol (TC) and low-density lipoprotein (LDL-C), increase high-density lipoprotein (HDL-C), obviously reduce serum ALT and AST levels, and show good blood fat reducing effect and liver protection effect.
The animal model of non-alcoholic fatty liver induced by high-sugar and high-fat feeding shows that the inonotus obliquus polysaccharide can reduce Total Cholesterol (TC) and low-density lipoprotein (LDL-C) in mouse serum and inhibit liver damage.
Acute alcoholic liver injury model experiments show that the inonotus obliquus polysaccharide can reduce the activity of AST and ALT in rat serum, relieve liver degeneration and inflammatory reaction, and show a certain liver protection effect.
Homocysteine is an important disease independent risk factor and is involved in the occurrence and development of various cardiovascular diseases and steatohepatitis. However, no report on the reduction of serum homocysteine by inonotus obliquus is available at present. Although there are reports that blood lipid and homocysteine levels in the serum of some cardiovascular disease patients are in positive correlation, there is no report that the two are necessarily correlated, and there is no report that lipid-lowering drugs have the effect of reducing homocysteine.
Disclosure of Invention
The invention aims to provide a new application of inonotus obliquus extract in reducing homocysteine.
Specifically, the invention relates to an application of inonotus obliquus extract as a pharmaceutical active ingredient in preparation of a medicament and/or a functional food for treating or preventing hyperhomocysteinemia.
Furthermore, the invention also relates to the application of the inonotus obliquus extract as a medicinal active ingredient in preparing medicines and/or functional foods for reducing homocysteine in hyperhomocysteinemia serum.
Furthermore, the invention also relates to the application of the inonotus obliquus extract as a medicinal active ingredient in preparing medicines and/or functional foods for reducing homocysteine in serum accompanied with cardiovascular diseases.
The inonotus obliquus extract is polysaccharide and glycoprotein active ingredients extracted from inonotus obliquus by a water extraction and alcohol precipitation method.
Specifically, the present invention adopts the following method to extract the inonotus obliquus extract from the inonotus obliquus.
Adding water with the mass being 8-10 times that of the inonotus obliquus into the inonotus obliquus, soaking for 12 hours, performing reflux extraction for 1.5-2.5 hours, filtering, and collecting filtrate; adding water with the mass 5-7 times that of the inonotus obliquus into the medicine residues, performing reflux extraction for 1-2 hours, filtering, and collecting filtrate; combining the filtrates obtained by the two-time collection, concentrating, and adding 95% (V/V) ethanol until the concentration of the ethanol in the concentrated solution is 65-80% (V/V); standing at room temperature for precipitating with ethanol for 12 hr, centrifuging to collect precipitate, dissolving with water, filtering, and freeze drying to obtain Inonotus obliquus extract.
Further, the inonotus obliquus extract is used as a pharmaceutical active ingredient, and pharmaceutically acceptable pharmaceutical carriers or excipients which can be used in combination with the active ingredient are added to prepare various pharmaceutically common oral dosage forms such as powder, granules, tablets, dripping pills, capsules and the like.
Furthermore, the invention can also use the inonotus obliquus extract as a functional additive to be added into various foods to prepare functional foods, wherein the foods comprise but are not limited to steamed bread, cakes, biscuits or instant noodles and the like.
According to the invention, a methionine feed is constructed to feed a rat model inducing hyperhomocysteinemia, and different doses of inonotus obliquus extract are administered by intragastric administration, so that the inonotus obliquus extract has the obvious effect of reducing the concentration of homocysteine in serum, and the important application prospect of the inonotus obliquus extract in the aspects of preventing and treating hyperhomocysteinemia is shown.
Meanwhile, acute toxicity tests show that the inonotus obliquus extract is in a non-toxic level and has obvious advantages compared with similar therapeutic drugs.
Drawings
FIG. 1 is a NMR spectrum of Inonotus obliquus extract.
Detailed Description
The following examples are only preferred embodiments of the present invention and are not intended to limit the present invention in any way. Various modifications and alterations to this invention will become apparent to 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.
Example 1: preparation of Inonotus obliquus extract.
Inonotus obliquus (Inonotus obliquus) is collected from Shanxi Lu shan area and identified as Inonotus obliquus (Inonotus obliquus) of Polyporaceae.
Soaking Inonotus obliquus in 10 times of water for 12 hr, heating under reflux for 2.5 hr, filtering, and collecting filtrate. Adding water with the mass 6 times of that of Inonotus obliquus into the residue, reflux-extracting for 1.5 hr, filtering, and collecting filtrate.
Mixing the two collected filtrates, concentrating to 1/5 volume of the filtrate, adding 95% ethanol, adjusting ethanol concentration of the concentrated solution to 70%, standing at room temperature, and precipitating with ethanol for 12 h.
Centrifuging, collecting precipitate, dissolving the precipitate with water, filtering, and freeze drying the filtrate to obtain Inonotus obliquus extract.
NMR spectrum of Inonotus obliquus extract(s) ((1H-NMR) is shown in FIG. 1, and its main component is polysaccharide.
Example 2: acute toxicity test of Inonotus obliquus extract.
Experimental ICR mice were randomly divided into 7 groups of 10 mice per group, each half male and female, and separately housed according to initial body weight.
According to the Hohner method, the test samples were divided into a blank control group, Inonotus obliquus extract groups 1, 2, 3, 4 and 5, and an Inonotus obliquus extract maximum tolerance group.
Wherein, the feeding dosages of the groups of the inonotus obliquus extract 1, 2, 3, 4 and 5 are respectively 1.00, 2.15, 4.64, 10.00 and 12.15g/kg BW, and the feeding dosage of the group with the maximum tolerance of the inonotus obliquus extract is 22g/kg BW.
After all mice are fed adaptively, the mice are fasted and are not forbidden to be fed with water for more than 12 hours, the inonotus obliquus extract is orally administrated with intragastric administration once according to the corresponding dose, and the blank control group is administrated with a certain amount of normal saline.
After 2h of administration, the mice were kept free to eat and drink water, and closely observed for all mice for their behavioral activities, coat color, mental status, stool status, drinking and eating, and other toxic reactions and death status. The body weight was weighed at intervals of 5 days, 10 days, and 14 days.
After the observation for 14 days, the mice are fasted without water supply for more than 12 hours, all the mice are killed after neck removal, autopsy is carried out, each tissue position is checked, and whether lesions exist or not is observed. The heart, liver, spleen, lung and kidney tissues are taken, weighed and the organ coefficient of each tissue is calculated.
Acute toxicity test results show that the inonotus obliquus extract has no influence on the body weight, heart, liver, spleen, lung and kidney organ coefficients of mice at the maximum tolerated dose of 22g/kg, and has no death phenomenon, which indicates that the extract is in a non-toxic level.
Example 3: and (3) carrying out a serum homocysteine concentration reduction test on the inonotus obliquus extract.
SD rats were taken and, after 1 week of adaptive feeding, randomly divided into 5 groups of a normal control group (n =8), a model control group (n =8), a folic acid control group (n =8), a low concentration group of inonotus obliquus extract (400mg/kg, n =8) and a high concentration group of inonotus obliquus extract (800mg/kg, n = 8).
The rats in the control group were fed with normal feed and the other 4 groups were fed with feed containing 3% methionine for 8 weeks. Therapeutic intervention is then initiated.
The folic acid control group is administered folic acid 4mg/kg per day by intragastric administration, the Fuscoporia obliqua extract low concentration group and Fuscoporia obliqua extract high concentration group are administered Fuscoporia obliqua extract 400mg/kg and 800mg/kg by intragastric administration, and the kit detects serum homocysteine level after continuous intervention for 4 weeks.
The serum homocysteine-reducing results of the inonotus obliquus extract of the invention on a rat experimental model for feeding and inducing hyperhomocysteinemia by aiming at 3% methionine feed are listed in table 1.
In table 1, compared to the normal control group:** p <0.01; comparison with model control group:## p < 0.01。
test results show that the concentration of homocysteine in the serum of rats in the model control group after molding is more than 15 mu mol/L, which indicates that the molding is successful. The inonotus obliquus extract 400mg/kg and 800mg/kg both have the effect of remarkably reducing homocysteine in the serum of a rat with hyperhomocysteinemia induced by methionine feed feeding, wherein the effect of 800mg/kg in a high-dose group is not different from the effect of positive drug folic acid, and the effect is equivalent. Compared with folic acid, the inonotus obliquus extract has no toxic or side effect and has obvious advantage in application of reducing homocysteine.
Claims (3)
1. Application of Inonotus obliquus extract in preparing medicine and/or functional food for treating or preventing hyperhomocysteinemia is provided.
2. Application of Inonotus obliquus extract in preparing medicine and/or functional food for reducing homocysteine in hyperhomocysteinemia serum is provided.
3. Application of Inonotus obliquus extract in preparing medicine and/or functional food for reducing homocysteine in blood serum of cardiovascular diseases accompanied with hyperhomocysteinemia is provided.
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Citations (3)
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|---|---|---|---|---|
| CN110430865A (en) * | 2016-12-21 | 2019-11-08 | 阿克生物科技公司 | Cosmetic composition and application thereof comprising Inonotus obliquus extract |
| CN110585321A (en) * | 2019-10-14 | 2019-12-20 | 黑龙江中医药大学 | Traditional Chinese medicine composition for preventing and treating anemia and preparation method and application thereof |
| US20200000136A1 (en) * | 2017-02-28 | 2020-01-02 | Cg-Bio Genomics, Inc. | Healthful supplement food |
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| CN110430865A (en) * | 2016-12-21 | 2019-11-08 | 阿克生物科技公司 | Cosmetic composition and application thereof comprising Inonotus obliquus extract |
| US20200000136A1 (en) * | 2017-02-28 | 2020-01-02 | Cg-Bio Genomics, Inc. | Healthful supplement food |
| CN110585321A (en) * | 2019-10-14 | 2019-12-20 | 黑龙江中医药大学 | Traditional Chinese medicine composition for preventing and treating anemia and preparation method and application thereof |
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