CN107496435B - Application of Anoectochilus formosanus glycoside in preparation of anti-fatigue medicine - Google Patents
Application of Anoectochilus formosanus glycoside in preparation of anti-fatigue medicine Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
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- 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
- 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
Abstract
The invention relates to the technical field of medicines, in particular to application of anoectochilus formosanus glycoside in preparing anti-fatigue medicines or foods. The invention provides a new medical application of the kinsenoside, and pharmacodynamic studies show that the kinsenoside can obviously prolong the swimming time of loaded mice, has obvious anti-fatigue effect, and can be used for preparing anti-fatigue medicaments or foods. The invention provides a new source for seeking new anti-fatigue drugs.
Description
Technical Field
The invention relates to the technical field of medicines, in particular to application of Anoectochilus roxburghii glycoside prepared from plants of the genus Kaliptae and the kindred plants thereof in anti-fatigue medicines or foods.
Background
Fatigue is a physiological and psychological phenomenon caused by excessive mental and physical activities of the body, mental stimulation and other factors. Generally, it can be classified into peripheral fatigue and central fatigue. With the development of economy, the rhythm of life is accelerated, and people working for a long time cannot effectively sleep or have irregular life, so that more and more people feel tired, muscle pain, inattention, memory loss, low mood, sleep disorder and the like. These symptoms, clinically commonly diagnosed as fatigue, encompass peripheral and central fatigue, with peripheral fatigue often occurring in physical workers and central fatigue often occurring in mental workers. Fatigue, especially central fatigue, can be converted into chronic fatigue and even 'fatigue death' if the central fatigue cannot be effectively treated for a long time, and the daily life and work of a patient are seriously interfered. The existing clinical anti-fatigue drugs are strictly limited due to adverse reactions such as side effects, addiction, excitability and the like. Therefore, the search for anti-fatigue drugs with high efficiency and low toxicity is still a difficult task and problem faced by the world medical community.
Anoectochilus roxburghii is a characteristic Chinese herbal medicine of Fujian, is derived from Anoectochilus Blume plant leaves and Laeliocattleya patens of Orchidaceae (Orchidaceae), is used as a medicine by the whole herb, has the effects of clearing heat and cooling blood, dispelling wind and promoting diuresis, detoxifying and the like (Fujian Chinese herbal medicine standard, Fuzhou: Haifeng Press, 2006: 154), is mainly used for treating hypertension, diabetes, hepatitis, tumors and the like, and has excellent curative effect, so the Anoectochilus roxburghii has the reputations of medicine king, gold grass and the like in folk. Except for Kalipira floribunda, various congeneric and kindred plants such as Taiwan chloranthus glaber, Xingren Anoectochilus roxburghii and beautiful anoectochilus roxburghii are frequently used as anoectochilus roxburghii medicinal materials in folk (Zhang Fe, Wanjing, Muhui Jianhua, preliminary investigation of Anoectochilus roxburghii germplasm resources in Shang region, academic report of Special schools such as Shang Shi et Shen et high, 2005,18(1): 26-28; Zheng pure, Huang Fang, Jilian Fang, literature examination certificate of Anoectochilus roxburghii, original plant and commodity investigation, Chinese herbal medicine 1996,27(3):169 one 171.). The herba Anoectochili Roxburghii medicinal material mainly contains lactone glycosides (such as herba Anoectochili Roxburghii glycoside), polysaccharides and flavonoids, especially contains high content of herba Anoectochili Roxburghii glycoside and polysaccharides. Modern pharmacological studies have shown that Anoectochilus roxburghii glycoside is one of the most active ingredients of Anoectochilus roxburghii, having immunomodulatory effects (Xiaoang M, Liu T, Tan W, Ren H, Li H, Liu J, Cao H, Cheng Q, Liu X, Zhu H, Tuo Y, Wang J, Zhang Y. effects of kinsenoside, a potential immunological delivery drug for autoimmunity drugs, on cognitive cells/CD8+ T cell communication in mice, Heaptology.2016 (64) (6): 2135. HARA 2150.), hypoglycemic effects (Zhang Y, Cai J, Ruan H, Pi H, Wu J. anatomical compliance of liver, a tissue culture, polysaccharide J. polysaccharide D.135, polysaccharide J.A. polysaccharide D.A. cholesterol, polysaccharide J. polysaccharide D.A. cholesterol, polysaccharide D.A. cholesterol, polysaccharide D.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.B.A.A.A.A.A.A.B.A.A.A.A.A.B.A.A.A.A.A.A.B.A.A.A.A.A.A.A.A.A.A.B.A.A.A.A.A.B.A.A.B.B.A.A.A.A.B.B.A.A.A.A.A.A.B.A.B.B.A.A.A.A.A.B.B.B.A.A.A.A.B.B.A.A.A.B.A.A.A.A.A.A.A.A.A.A.A.A.B.B.A.B.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.A.B.A.A.A.A.A.B.B.A.A.A.A.A.A.B.A.B.A.B.B.B.A.B.B.B.A.B.B.B.B.B.A.A.A.B.B.A.A.A.B.B.A.A.A.A.A.A.A.B.A.A.A.A.A.A.A.A.A.A.A.A.B.A.A.A.A.A.B.B.B.B.B.A.A.B.B.A.B.B.A.B.B.A.A.B.B.B.B.B.B.B.B.A.B.B.B.B.B.B.B.B.B.B.B.B.B.B.B.A.B.B.A.A.A.A.A.A.A.B.B.B., Hypolipidemic (Rehman SU, Choi MS, Kim IS, Luo Z, Xue Y, Yao G, Zhang Y, Yoo HH. in Vitro Association of CYP-Mediated Drug Interactions for kinase, an anti-inflammatory peptide of molecular kinase 2016,21(6) pii: E800.), anti-inflammatory (Hsiao HB1, Hsieh CC, Wu, Lin H1, Lin WC. kinase inhibitors of collagen promoter type-II collagen induced expression of molecular kinase, bone regulation peptide of BMC, skeleton Alternan, 2016, 80, and anti-bone activity of bone graft peptide of kinase, collagen promoter, skeleton promoter, collagen promoter, collagen promoter, collagen promoter, collagen, however, reports on the use of Anoectochilus roxburghii and Anoectochilus roxburghii glycoside as a monomer compound for resisting fatigue are not found so far.
Disclosure of Invention
The invention aims to provide a new medical application of the Anoectochilus roxburghii glycoside.
In a first aspect of the invention, the application of the Anoectochilus roxburghii glycoside in preparing an anti-fatigue medicine or food is provided.
The chemical structure of the anoectochilus formosanus glycoside is shown as follows:
in a second aspect of the invention, an anti-fatigue drug or food is provided, wherein the active ingredient in the anti-fatigue drug or food is the roxburgh anoectochilus terminal bud glycoside.
Preferably, the anti-fatigue medicament or food further comprises pharmaceutically acceptable auxiliary materials.
The preparation method of the Anoectochilus roxburghii glycoside comprises the following steps:
(A) preparing an extracting solution: crushing medicinal materials of the plants of the genus Labiatae and the kindred plants thereof, putting the crushed medicinal materials into an extraction tank, carrying out hot extraction for 2-3 times by using 40-90% ethanol, wherein the using amount of a solvent is about 8-10 times of the crude drug amount each time, and the extraction time is 1-2 hours each time, and combining extracting solutions;
(B) refining, concentrating and drying: concentrating the extracting solution obtained in the step A, extracting the concentrated solution by ethyl acetate to remove impurities, and concentrating and drying a water layer to obtain a crude extract;
(C) separation and purification: and B, dissolving the crude extract obtained in the step B in water, loading the crude extract into a macroporous adsorption resin, collecting 0-10% ethanol elution part, concentrating and drying to obtain a crude product of the kinsenoside, loading the sample into a forward silica gel column, eluting by adopting a dichloromethane-methanol system, and collecting 6:1 elution part to obtain the kinsenoside (the purity is more than 90%).
Preferably, the labiate and its kindred plants in step a are selected from one or more of liptae mosaic, Taiwan silver orchid, Zhejiang gold orchid, Xinnao gold orchid, Yunnan gold orchid, long-sheet gold orchid, Plumbum gold orchid, Lily bud gold orchid, Goodyera anoectochilus, Anoectochilus labris, Spotted leaf orchid, big flower spot leaf orchid, small spot leaf orchid, Maochytenus glabra, Tianquan spot leaf orchid and Xueyleaf orchid.
Preferably, the concentration in step B is a recovery of the solvent under reduced pressure.
Preferably, the macroporous resin in step C is selected from one of HP-20, AB-8, D-101, ZTC-1, HPD-100 or DA 201.
The invention adopts a load-bearing Mouse forced swimming model (Liu Y, Li L, An S, Zhang Y, Feng S, ZHao L, Teng L, Wang D. anti facial Effects of anti-classic cornea A Current Cultured Mycelium vita modulus of Oxidative Stress signalling in a Mouse model. biomed Res int.2017:9374026.) to observe the anti-fatigue effect of the perianthrin intragastric administration provided by the invention. The animal model is a widely accepted anti-fatigue animal model, and experiments mainly aim at the exercise-induced fatigue, including central fatigue and peripheral fatigue (Wang jun, Zhou Jun, some thinking of the animal model in the research of the exercise-induced fatigue of traditional Chinese medicine, China journal of experimental prescriptions, 2009,15(3): 83-85; Zheng lan, Lu Aiyun, the research of the exercise-induced fatigue animal model, China sports science and technology, 2003,39(2): 20-23; Tianfeng, recognitions of the central and peripheral exercise-induced fatigue, China tissue engineering research, 2015,19(42): 6849-6854).
The experimental result shows that after the continuous gavage administration of 100mg/kg of the anoectochilus formosanus glycoside for 18 days, the normal physiological condition and the weight change of a mouse are not obviously influenced; the weight swimming time of the mice in the blank group is 1577.7 +/-303.1 seconds, the weight swimming time of the mice in the anoectochilus formosanus glycoside administration group is 2775.2 +/-770.3 seconds, and the significant difference (p is less than 0.01) exists between the mice in the blank group and the mice in the anoectochilus formosanus glycoside administration group; furthermore, the kinsenoside can also obviously reduce the BLD (lactic acid) content in the serum of a load bearing swimming mouse and improve the activity of LDH (lactate dehydrogenase) in the serum of the mouse, which shows that the kinsenoside has good anti-fatigue effect.
The anti-fatigue of the invention means preventing or treating peripheral fatigue or central fatigue, improving the working capacity of the organism, reducing the content of lactic acid in serum, improving the activity of lactate dehydrogenase in serum and the like, and improving symptoms of muscle pain, attention deficit, memory decline and the like.
The invention finds an ideal anti-fatigue medicine. The invention provides a new medical application of the kinsenoside, and pharmacodynamic studies show that the kinsenoside can obviously prolong the swimming time of loaded mice, has obvious anti-fatigue effect, and can be used for preparing anti-fatigue medicaments or foods. The invention provides a new source for seeking new anti-fatigue drugs.
Drawings
FIG. 1 is a graph of animal body weight over time;
fig. 2 is the effect of nefarin on swimming time for weight bearing mice (mean ± SD, n ═ 10), where ×. vs. blank control, p < 0.01;
fig. 3 is the effect of kinsenoside on BLD (lactic acid) levels in serum of swimming-weight mice (mean ± SD, n ═ 10), where ×/vs blank control, p < 0.01;
fig. 4 is the effect of ectoin on LDH (lactate dehydrogenase) levels in serum of swimming-weight mice (mean ± SD, n ═ 10), where vs. blank control, p < 0.05.
Detailed Description
The following detailed description of the present invention will be made with reference to the accompanying drawings.
Example 1: preparation of Anoectochilus formosanus glycoside
(1) Preparing an extracting solution: crushing medicinal materials of the plants of the genus Labiatae and the kindred plants thereof, putting the crushed medicinal materials into an extraction tank, carrying out hot extraction for 2-3 times by using 40-90% ethanol, wherein the using amount of a solvent is about 8-10 times of the crude drug amount each time, and the extraction time is 1-2 hours each time, and combining extracting solutions;
(2) refining, concentrating and drying: concentrating the above extractive solution, extracting the concentrated solution with ethyl acetate to remove impurities, concentrating the water layer, and drying to obtain crude extract;
(3) separation and purification: dissolving the crude extract in water, loading the crude extract on macroporous adsorption resin, collecting 0-10% ethanol elution part, concentrating and drying to obtain a crude product of the kinsenoside, loading the sample on a positive silica gel column, eluting by adopting a dichloromethane-methanol system, and collecting 6:1 elution part to obtain the kinsenoside (the purity is more than 90%).
Example 2: anoectochilus formosanus glycoside anti-fatigue pharmacodynamic experiment
2.1 materials of the experiment
18-22 g ICR mice (provided by the experimental animal center of the second army medical university) 20 mice, and a mouse LDH (lactate dehydrogenase) and BLA (blood lactate) kit (Nanjing institute of bioengineering). The main apparatus is as follows: a 5L beaker (Shanghai cereal steam glass instruments, Inc.), a microplate reader (BIO-RAD 550), a centrifuge (Labofuge 400R, Heraeus), and an electronic balance (FA 110A, Shanghai precision scientific instruments, Inc.).
2.2 animal grouping and Experimental treatment
Before the experiment, 20 mice were weighed, and randomly divided into 2 groups according to body mass, 10 mice in each group, free diet and water intake. Is divided into blank group and anoectochilus formosanus glycoside group. Blank group: the same volume of physiological saline as the administration group was administered; anoectochilus formosanus glycoside group: the preparation is administered by intragastric administration once daily at a dose of 100mg/kg for 18 days.
2.3 Observation index
(1) The change in body mass of the mice in each group was recorded and weighed every 9 days.
(2) After the last day of intragastric administration for 30min, 6 percent of lead is loaded at the tail root of the mouse, and the mouse starts swimming in a 5L beaker until the head can not float out of the water surface within 7s, and the timing is stopped, and the exhaustion time is counted.
(3) The mouse is taken out, and the eyeball is pulled out to take blood. Centrifuging the blood at 4 deg.C and 3500r/min for 10 min, collecting the supernatant, and storing at-80 deg.C. BLA, LDH were determined by performing the procedures according to the kit instructions.
2.4 results of the experiment
Two-sample t-test statistical analysis was performed on both sets of data using SPSS13.0, and differences were considered statistically significant when P < 0.05.
(1) Body weight changes in mice
The body weight of the mice in the blank group gradually increased, the body weight of the mice treated by the anoectochilin administration was basically stable, and the difference from the blank group was not significant, which indicates that the anoectochilin had no influence on the normal physiological condition of the mice, as shown in fig. 1.
(2) Mouse weight bearing swimming time
As shown in figure 2, the negative gravity weight exhaustive swimming time of the mice in the blank group is 1577.7 +/-303.1 seconds, the negative gravity weight exhaustive swimming time of the mice in the anoectochilus formosanus glycoside administration group is 2775.2 +/-770.3 seconds, and the significant difference (p <0.01) exists between the negative gravity weight exhaustive swimming time and the weight exhaustive swimming time of the mice in the blank group, which indicates that the anoectochilus formosanus glycoside can significantly increase the weight swimming time of the mice.
(3) Serum index
As shown in figures 3 and 4, compared with the blank group, the kinsenoside group has p <0.05, and the kinsenoside can obviously reduce the content of lactic acid BLA in the serum of mice and improve the activity of lactate dehydrogenase LDH in the serum of the mice, so that the kinsenoside 100m g/kg administered by intragastric administration can obviously improve the level of fatigue-related factors in the serum of the animal model of the weight-bearing mice.
In view of the pharmacodynamic experimental results, the Anoectochilus roxburghii glycoside has obvious anti-fatigue activity, so that the Anoectochilus roxburghii glycoside can be used for preparing anti-fatigue medicines or foods.
While the preferred embodiments of the present invention have been described in detail, it will be understood by those skilled in the art that the invention is not limited thereto, and that various changes and modifications may be made without departing from the spirit of the invention, and the scope of the appended claims is to be accorded the full range of equivalents.
Claims (2)
1. Application of Anoectochilus roxburghii glycoside in preparing antifatigue medicine or food is provided.
2. The preparation method of the anoectochilus formosanus glycoside is characterized by comprising the following steps:
(A) preparing an extracting solution: crushing medicinal materials of the plants of the genus Labiatae and the kindred plants thereof, putting the crushed medicinal materials into an extraction tank, carrying out hot extraction for 2-3 times by using 40-90% ethanol, wherein the using amount of a solvent is about 8-10 times of the crude drug amount each time, and the extraction time is 1-2 hours each time, and combining extracting solutions; the labiate and the kindred plants thereof are selected from one or more than two of mosaic labiate, Taiwan silver line orchid, Zhejiang gold line orchid, Xingren gold line orchid, Yunnan gold line orchid, long-sheet gold line orchid, Plumbum gold line orchid, beautiful bud gold line orchid, high gold line orchid, short lip gold line orchid, spotted leaf orchid, large spotted leaf orchid, small spotted leaf orchid, Maotai spotted leaf orchid, Tianquan spotted leaf orchid and Xueyleaf orchid;
(B) refining, concentrating and drying: concentrating the extracting solution obtained in the step A, extracting the concentrated solution by ethyl acetate to remove impurities, and concentrating and drying a water layer to obtain a crude extract; the concentration is to recover the solvent under reduced pressure;
(C) separation and purification: b, dissolving the crude extract obtained in the step B in water, loading the crude extract into macroporous adsorption resin, collecting 0-10% ethanol elution part, concentrating and drying to obtain a crude product of the kinsenoside, loading the sample into a forward silica gel column, eluting by adopting a dichloromethane-methanol system, and collecting 6:1 elution part to obtain the kinsenoside; the macroporous resin is selected from one of HP-20, AB-8, D-101, ZTC-1, HPD-100 or DA 201.
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| CN112843177A (en) * | 2021-02-24 | 2021-05-28 | 中国人民解放军海军军医大学 | Application of anoectochilus formosanus extract in preparation of medicine or health-care product for treating or relieving pain |
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