CN111803591A - Application of dry dendrobium aqueous extract in preparation of obesity treatment drug - Google Patents
Application of dry dendrobium aqueous extract in preparation of obesity treatment drug Download PDFInfo
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- CN111803591A CN111803591A CN202010793227.1A CN202010793227A CN111803591A CN 111803591 A CN111803591 A CN 111803591A CN 202010793227 A CN202010793227 A CN 202010793227A CN 111803591 A CN111803591 A CN 111803591A
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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)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/898—Orchidaceae (Orchid family)
- A61K36/8984—Dendrobium
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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
- A61P3/04—Anorexiants; Antiobesity agents
Abstract
The invention belongs to the technical field of pharmacy, and relates to application of a dried dendrobium aqueous extract in preparation of a medicament for treating obesity. In particular to a dendrobium stem aqueous extract which has obvious inhibition effect on the synthesis of triglyceride under the high-fat state. Experimental results show that the dry dendrobium aqueous extract prepared by the invention can obviously inhibit the gene expression of fatty acid binding protein, further has a beneficial effect on fatty acid metabolism and inhibits the occurrence of obesity diseases, can obviously inhibit the gene expression of diacylglycerol acyltransferase in HUH7 cells, inhibits the synthesis of triglyceride, increases the glucose uptake rate and energy consumption, and further resists obesity caused by high-fat diet, so that the dry dendrobium aqueous extract prepared by the invention can be applied to the preparation of medicines for treating obesity.
Description
Technical Field
The invention relates to the technical field of pharmacy, in particular to application of a dried dendrobium aqueous extract in preparing a medicament for treating obesity.
Background
Obesity and its associated metabolic diseases are becoming one of the important threats affecting human health worldwide. Further, modern people have an increased visceral fat obesity accompanying changes in living environment, and the onset of metabolic syndrome (metabiolsyndrome) associated with diabetes, hypertension, lipid metabolism disorders, insulin resistance, and the like has been rapidly increased, and these diseases are associated with an increased risk of occurrence of each other, and are common diseases associated with diversified metabolic changes of living bodies such as aging, stress, and immune function reduction. As a typical conventional obesity therapeutic agent, sunitinib, which is a main raw material of Orlistat (Orlistat), has side effects such as diarrhea, flatulence, and abdominal distension, and nominatine has side effects such as headache, thirst, anorexia, insomnia, and constipation. Also, orlistat inhibits the absorption of vitamin E and vitamin D, phentermine and sibutramine have the side effects of causing heart rate increase/palpitation, dizziness. Therefore, with the limitation of western medicine in overcoming the side effects and chronic diseases of synthetic pharmaceuticals, new natural drugs are being developed.
Dendrobe is a common and rare traditional Chinese medicine, is sweet, light and slightly salty in taste and cold, and enters stomach, kidney and lung meridians. The dendrobium huoshanense, the dendrobium officinale, the dendrobium nobile and the dendrobium nobile are superior products in dendrobium nobile. Also called Linlan (Shennong Bencao Jing), Qiannian Ruan (Bencao gang mu), etc., it has higher ornamental value and medicinal function, recorded in Li Shizhen Bencao gang mu, dendrobe has the efficacies of nourishing five internal organs, fatigue slimming, nourishing yin and replenishing essence, taking thick intestines and stomach for a long time, calming stomach qi, growing muscle, fixing mind and removing fright, reducing weight and prolonging life, etc. According to the research of our literature, the dendrobium or the dendrobium extract has not been found to be applicable to the treatment of obesity diseases.
Disclosure of Invention
The invention aims to provide an application of a dry dendrobium aqueous extract in preparing a medicine for treating obesity, and is a new application of the dendrobium extract. At present, no report indicates that the dried dendrobium water extract can be applied to the treatment of obesity.
The application of the dry dendrobium aqueous extract in preparing the medicine for treating obesity is characterized in that the dry dendrobium aqueous extract is obtained by extracting dendrobium medicinal materials.
Further, the dendrobium stem is one or a combination of more than two of dendrobium huoshanense, dendrobium officinale, dendrobium nobile, dendrobium fimbriatum and dendrobium chrysotoxum.
Further, the preparation steps of the dried dendrobium aqueous extract are as follows: extracting herba Dendrobii dry powder with 18-20 times of distilled water in 55-65 deg.C water bath, filtering, collecting filtrate, repeatedly extracting, mixing extractive solutions, concentrating under reduced pressure to 1/6-1/8 of original total volume, and freeze drying to constant weight.
Furthermore, the dried dendrobium aqueous extract can be prepared into any pharmaceutically acceptable oral and non-oral preparation forms, such as one of granules, pills, capsules, tablets, powder, paste and oral liquid.
The invention has the beneficial effects that:
(1) the dry dendrobium aqueous extract prepared by the invention can obviously inhibit the gene expression of fatty acid binding protein, has beneficial effect on fatty acid metabolism, and further inhibits the occurrence of obesity;
(2) the dry dendrobium aqueous extract prepared by the invention can obviously inhibit gene expression of diacylglycerol acyltransferase in HUH7 cells, inhibit synthesis of triglyceride, increase glucose uptake rate and energy consumption, and further resist obesity caused by high-fat diet.
Therefore, the dry dendrobium aqueous extract prepared by the invention can be used for preparing a medicament for treating obesity and has good development prospect in the aspect of treating obesity diseases.
Drawings
FIG. 1 shows that the dry dendrobium aqueous extract inhibits the gene expression of fatty acid binding protein;
wherein, the data are expressed as means ± SD, FABP 4: the fatty acid binding protein4 is a protein that binds to fatty acids,**P＜0.01，***P<0.05; administration group vs blank pairGroup control; DWE: dry Dendrobium Water extract of Dendrobium officinale; fold: multiple times.
FIG. 2 shows that an aqueous extract of dried Dendrobium nobile inhibits the gene expression of diacylglycerol acyltransferase in HUH7 cells;
where data are expressed as means ± SD, DGAT 1: (ii) a diacylglycerol acyltransferase 1,**P＜0.01，***P<0.05; a administration group vs blank group; DWE: dry Dendrobium Water extract of Dendrobium officinale; fold: the number of times of the multiple,
FIG. 3 is a scheme of triglyceride synthesis.
Detailed Description
The present invention will be further illustrated with reference to the following specific examples.
Triglyceride (abbreviated as TG) is the main form of biological energy storage, when the energy taken in by human body exceeds the energy consumed by human body, the surplus energy is stored in the form of T G in fat tissue, and meanwhile, the excessive TG can be deposited in the tissues such as islet beta cells, liver and the like, and finally, the diseases such as obesity, diabetes and the like are caused. Triglyceride (TG) is the major biological energy storage in vivo, glycerol TG is synthesized primarily in the liver, adipose tissue, and small intestine, and there are two pathways for TG synthesis (see fig. 3): the 3-phosphoglycerate pathway (major pathway) and the monoacylglycerol pathway (minor pathway). First, glycerol-3-phosphate reacts with fatty acyl-CoA to produce phosphatidic acid, which is dephosphorylated by the action of a phosphatase to form Diacylglycerol (DAG), which is also produced via the secondary pathway monoacylglycerol and fatty acyl-CoA. DAG catalyzed TG synthesis last step: transfer of acyl-CoA acyl groups to DAG to form TG is the rate-limiting enzyme in TG synthesis. When the human body takes more energy than it consumes, the excess energy is stored in the adipose tissue in the form of TG, and at the same time, the excess TG is deposited in the liver tissue, eventually leading to obesity. Therefore, inhibition of TG synthesis in the above tissues can be used as a strategy for the treatment of obesity.
Diacylglycerol acyltransferase (DGAT) is a key enzyme catalyzing TG synthesis pathway, and the level of expression directly affects the final TG content. DGAT is an endoplasmic reticulum membrane integrin and has two subtypes: the two types of DGAT1 and DGAT2 show that the research shows that the inhibition of the activity of DGAT1 can reduce the synthesis of TG, improve the sensitivity of the body to insulin and leptin, increase the glucose uptake rate and energy consumption, resist obesity caused by high fat diet, and the like, and can be used as a target for treating obesity diseases.
Obesity is characterized by hyperplastic hypertrophy of adipocytes, which are now thought to secrete large amounts of biologically active substances associated with systemic islet cord sensitivity and energy metabolism, with accompanying infiltration and accumulation of macrophages in adipose tissue. Fatty acid binding proteins (fattyacid-binding protein4, FABP4) are secreted from adipocytes and are present in large amounts in human serum. FABP4 is a family of endogenous low molecular weight intracellular proteins, widely distributed in various cells of the small intestine, liver, fat, heart, brain and skeletal muscle of mammals, and has tissue specificity. FABP4 plays an important role in the regulation of the uptake, transport and metabolism of long chain fatty acids. The research result of FABP4 gene-deficient mice shows that the reduction of the expression of FABP4 can improve the peripheral insulin resistance, protect the function of islet beta cells, and produce beneficial effects on fatty acid metabolism, thereby inhibiting the occurrence of obesity, and therefore, the FABP4 gene-deficient mice can be used as a targeted drug for treating obesity.
According to the invention, the mRNA expression level of the TG synthesis and lipid metabolism related genes is detected, so that the effect of the dendrobium stem aqueous extract as a medicament applied to obesity treatment is researched.
Because the existing medicinal dendrobium has various types, the embodiment of the invention takes dendrobium huoshanense as an example, and researches the effect of the dried dendrobium aqueous extract as a medicament applied to obesity treatment. However, the dendrobium in the invention is not limited to dendrobium huoshanense, but also can be medicinal dendrobium of other same genus, such as dendrobium officinale, dendrobium nobile, dendrobium fimbriatum or dendrobium chrysotoxum.
The dendrobium huoshanense is originally produced in places such as Anhuoshan in China, is the first protective variety in more than 200 endangered rare varieties which are important special items of national modern traditional Chinese medicines, is an extreme product in medicinal dendrobium, is also a unique and famous traditional Chinese medicine material with geographical marks of original production places in Hohan county, and is historically known as the first of nine major Chinese immortals and the health soft gold, and is recorded in Shen nong Ben Cao and Ben Cao gang mu. The polysaccharide contained in the dendrobium huoshanense can greatly improve SOD (main substance for delaying senility) in a human body, and has obvious effects on enhancing immunity, resisting fatigue, delaying senility, promoting cancer cell apoptosis and the like, thereby achieving the effects of health care and life prolonging.
Example 1
The preparation method of the dried dendrobium aqueous extract comprises the following specific steps:
preparing a dried dendrobium aqueous extract: weighing 20g of dendrobium huoshanense dry powder, extracting with 400ml of distilled water in a water bath at 55 ℃, filtering, collecting filtrate, repeatedly extracting for 3 times, extracting for 1 hour each time, and combining the extracting solutions for 3 times. Concentrating the combined extractive solutions at 55 deg.C under reduced pressure to 1/6-1/8 of original liquid volume, and freeze drying to constant weight to obtain dried herba Dendrobii water extract with mass volume concentration of 100 mg/ml.
The dry dendrobium aqueous extract with the mass concentration of 100 mg/mL prepared in the example 1 is filtered and sterilized by a filter membrane with the thickness of 0.22 mu m before the test, and then diluted to prepare a series of dry dendrobium aqueous extract liquid medicines with the mass concentrations of 10 mu g/mL, 100 mu g/mL and 1000 mu g/mL in sequence, and the dry dendrobium aqueous extract liquid medicines are refrigerated and stored for the test for standby.
(I) detecting the expression of mRNA of lipid metabolism related gene FABP4 in RAW264.7 cells by the dried dendrobium water extract
RAW264.7 (mouse mononuclear macrophage) DMEM complete medium (with 10% fetal calf serum, 100U/mL penicillin, 100. mu.g/mL streptomycin added) at 37 ℃ with 5% CO by volume fraction2The cells were synchronized for 12h and randomly divided into 2 groups, namely: blank control group: no drug administration (0. mu.g/ml); (ii) administration group: high dose group of dried herba Dendrobii water extract (DWE: 1000 μ g/mL), medium dose group of dried herba Dendrobii water extract (DWE: 100 μ g/mL), and low dose group of dried herba Dendrobii water extract (DWE: 10 μ g/mL); dosing for 12h, fluorescent quantitative PCR: total RNA from RAW264.7 cells was extracted using RNAioso Plus, 1. mu.g of total RNA was reverse transcribed to cDNA by Oligo dT18, and primers were PCR amplified by quantitative PCR technique using Toyobo company Thunde BirdSYBR qPCR Mix reagent as per instructionsAnd (5) increasing. Each set of samples had 3 duplicate wells to ensure validity of experimental data. Using GAPDH as an internal control, the change in expression of the target gene FABP4 (fatty acid binding protein) mRNA relative to the internal control Gene (GAPDH) was detected for quantification. Data (i.e., fold change in expression of the gene of interest in the experimental group relative to the blank control group) was analyzed using Graphpad 7. The primers used were: FABP4 (mFABP 4-F: AAATCACCGCAGACGACAGG; mFABP 4-R: AACTCTTGTGGAAGTCACGCC).
The experimental results shown in the attached figure 1 show that the dry dendrobium aqueous extract prepared by the invention can obviously inhibit the mRNA expression level of fatty acid binding protein (FABP4) by comparing with a blank control group (the administration concentration is 0 mug/mL), and the administration of the high-dose and medium-dose dendrobium aqueous extract can obviously inhibit the fatty acid binding protein (FABP4), so that the beneficial effect on fatty acid metabolism can be produced, and further the occurrence of obesity can be inhibited. Therefore, the dried dendrobium aqueous extract can be used as a medicine for treating obesity.
(2) Detecting the expression of the dendrobium stem aqueous extract on the mRNA of the TG synthesis related gene DGAT1 in vitro HUH7 cells
Grouping: after HUH7 cells were recovered, they were transferred to a petri dish with a high-fat (HFD) culture medium containing 40% HFD and 5% CO at 37 deg.C2Cultured under the condition, the cells are synchronized for 12h and randomly divided into 2 groups, namely: blank control group: no drug administration (0. mu.g/ml); (ii) administration group: a dry dendrobium aqueous extract high dose group (DWE: 1000 mug/mL), a dry dendrobium aqueous extract medium dose group (DWE: 100 mug/mL) and a dry dendrobium aqueous extract low dose group (DWE: 10 mug/mL); after 12h of dosing, fluorescent quantitative PCR: total RNA from HUH7 cells was extracted using RNAiso Plus, 1. mu.g of total RNA was reverse transcribed to cDNA by Oligo dT18, and primers were PCR amplified by quantitative PCR technique using Toyobo company ThunderBirdSYBR qPCR Mix reagent as per instructions. Each set of samples had 3 duplicate wells to ensure validity of experimental data. Using GAPDH as a reference, the change in expression of the gene of interest diacylglycerol acyltransferase 1(DGAT1) relative to the reference Gene (GAPDH) was detected for quantification. Data (i.e., fold change in expression of the gene of interest in the experimental group relative to the blank control group) was analyzed using Graphpad 7. The primers used were: DGAT1 (hDGAT 1-f: GGTGCCCTGACGGAGCAGGC;hDGAT1-r：CAGTAAGACCACAGCCGCTG）。
the experimental results shown in fig. 2 show that the dry dendrobium aqueous extract prepared by the invention can obviously inhibit the mRNA expression level of diacylglycerol acyltransferase 1(DGAT1) by administration of high-dose and medium-dose dendrobium aqueous extracts compared with a white control group (administration concentration is 0 μ g/mL), so that the dry dendrobium aqueous extract can inhibit the activity of DGAT1, and the reduction of the activity of DGAT1 can inhibit the synthesis of TG, thereby increasing the glucose uptake rate and energy consumption, and resisting obesity and the like caused by high-fat diet, and therefore, the dry dendrobium aqueous extract can be used as a medicament for treating obesity diseases.
In conclusion, the dendrobium stem aqueous extract provided by the invention can obviously inhibit the mRNA expression level of diacylglycerol acyltransferase (DGAT) and fatty acid binding protein (FABP4), not only can inhibit the synthesis of TG, but also can effectively regulate the level of lipid metabolism, so that the dendrobium stem aqueous extract can be used as a medicine for treating obesity.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (4)
1. The application of the dry dendrobium aqueous extract in preparing the medicine for treating obesity is characterized in that the dry dendrobium aqueous extract is obtained by extracting dendrobium medicinal materials.
2. The application of the dendrobium stem aqueous extract in preparing a medicine for treating obesity according to claim 1, wherein the dendrobium stem is one or a combination of more than two of dendrobium huoshanense, dendrobium officinale, dendrobium nobile, dendrobium fimbriatum and dendrobium chrysotoxum.
3. The application of the dry dendrobium aqueous extract in preparing the medicine for treating obesity according to claim 1, wherein the preparation steps of the dry dendrobium aqueous extract are as follows: extracting herba Dendrobii dry powder with 18-20 times of distilled water in 55-65 deg.C water bath, filtering, collecting filtrate, repeatedly extracting, mixing extractive solutions, concentrating under reduced pressure to 1/6-1/8 of original total volume, and freeze drying to constant weight.
4. The use of the aqueous extract of dried dendrobium nobile according to any one of claims 1 to 3 in the preparation of a medicament for the treatment of obesity, wherein the aqueous extract of dendrobium nobile can be prepared into all pharmaceutically acceptable oral and non-oral preparations.
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| US20220265754A1 (en) * | 2021-02-19 | 2022-08-25 | Multistars Biotechnology Company Limited | Method of improving diseases using traditional chinese medicine polysaccharides and bacterial composition thereof |
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Application publication date: 20201023 |