CN114128885A - Dendrobium compound preparation with functions of regulating immunity, promoting intestinal peristalsis and regulating intestinal flora and preparation method and application thereof - Google Patents
Dendrobium compound preparation with functions of regulating immunity, promoting intestinal peristalsis and regulating intestinal flora and preparation method and application thereof Download PDFInfo
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- CN114128885A CN114128885A CN202111418056.5A CN202111418056A CN114128885A CN 114128885 A CN114128885 A CN 114128885A CN 202111418056 A CN202111418056 A CN 202111418056A CN 114128885 A CN114128885 A CN 114128885A
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Abstract
The invention discloses a dendrobe compound preparation with the functions of regulating immunity, promoting intestinal peristalsis and regulating intestinal flora and a preparation method thereof, wherein the dendrobe compound preparation comprises the following components in parts by weight: 5-15 parts of dendrobium devoninum paxt, 2-10 parts of fructus cannabis, 2-8 parts of perilla seeds, 1-5 parts of poria cocos, 2-8 parts of Chinese dates and 2-8 parts of xylo-oligosaccharide. The preparation method of the dendrobium compound preparation comprises the main key process technologies of wet grinding of the purple dendrobium, dry grinding of fructus cannabis, perilla seeds, poria cocos and Chinese dates, torrent wall breaking extraction, plate and frame filter pressing, low-temperature concentration, freeze drying and the like, and various deep-processed products are prepared by adding auxiliary materials according to the needs. The herba Dendrobii compound preparation has effects of regulating immunity, promoting enterokinesia and regulating intestinal flora, and can be used for preparing functional food or medicine with immunity regulating and bowel relaxing effects.
Description
Technical Field
The invention relates to the technical field of dendrobe compound preparations, in particular to a dendrobe compound preparation with functions of regulating immunity, promoting enterokinesia and regulating intestinal flora, and a preparation method and application thereof.
Background
Immunity is a physiological function of the human body, by which the human body recognizes and destroys any foreign body (virus, bacteria, etc.) invaded from the outside, treats aged, damaged, dead, denatured self cells, and recognizes and treats mutant cells and virus-infected cells in the body. People with low immunity are easy to be infected, easily suffer from various diseases, are difficult to cure after repeated attack, and even induce serious diseases. Therefore, it is important to improve immunity for preventing various diseases and improving life quality, and it is considered as one of the best methods to improve immunity by taking functional foods or medicines having an immunoregulatory effect.
Constipation is a common complex symptom in clinic, and mainly refers to reduction of defecation frequency, reduction of stool quantity, dry and hard stool, difficult defecation and the like. Frequent constipation can cause adverse effects on human health, and at the moment, garbage in the intestinal tract cannot be timely excreted, so that the intestinal tract health environment can be possibly affected. The causes of long-term constipation are various, including the lack of beneficial bacteria in the body, which can cause damage to intestinal mucosa, damage to the intestinal environment, slow gastrointestinal peristalsis function, dryness of the intestinal tract and water shortage. The external manifestations are that the old excrement can not be discharged out of the body in time, the excrement is dry, the lower abdomen is prominent, the flatulence, the face is dark yellow, the mouth is dry and the halitosis is caused, and if the treatment is not carried out in time, the hemorrhoids are also caused. Therefore, the body should be regulated by proper diet to promote intestinal motility, regulate and improve intestinal flora, and prevent constipation.
In recent years, with the development of science and technology and the improvement of the living standard of Chinese people, the rational use of disease-preventing and body-building functional food has gradually become a conscious requirement of people, and the research and development in the field are more and more paid attention. The search for a functional food which is healthy, effective, free of toxic and side effects and has the functions of regulating immunity and relaxing bowel is a hot spot of current research and development.
Chinese patent application with application publication number CN 103719494B (application number 201310752822.0) discloses a burdock fruit health tea for relaxing bowels and moistening intestines and a preparation method thereof, belonging to the field of health products containing plant components for relaxing bowels and moistening intestines. The invention provides a burdock fruit health tea capable of relaxing bowels and moistening intestines, which comprises the following raw materials in parts by weight: 20-50 parts of burdock, 10-20 parts of artemisia anomala, 10-20 parts of radix rehmanniae, 10-20 parts of cortex lycii radicis, 10-20 parts of radix angelicae, 10-20 parts of poria cocos, 10-20 parts of salvia miltiorrhiza, 10-20 parts of codonopsis pilosula, 10-20 parts of eupatorium, 10-20 parts of gynostemma pentaphylla, 10-20 parts of white mulberry root-bark, 10-20 parts of trichosanthes kirilowii maxim, 10-20 parts of coix seed, 10-20 parts of perilla fruit, 10-20 parts of hawthorn, 10-20 parts of rhizoma alismatis and 10-20 parts of patrinia scabiosaefolia. Great burdock achene is rich in grease and is smooth and favorable in property. Modern pharmacological research shows that the main components of burdock are arctiin and fatty oil, and the components have the effects of relaxing bowels and moistening intestines. Clinical experience shows that the burdock is bitter and cool and has strong intestine smoothing and purgation effects, so that microcirculation can be improved for patients with constipation, and the burdock traditional Chinese medicine is light in taste, does not influence the taste of the burdock, is easy to accept by people and can better achieve the effects of relaxing bowels and moistening intestines.
The application publication number CN 109619487A (application number 201811272913.3) of Chinese invention patent application discloses a composition with the function of relaxing bowel and a preparation method thereof, relating to the technical field of health food. The invention discloses a composition with a bowel relaxing function and a preparation method thereof, and the composition with the bowel relaxing function comprises 4 parts of solid microcapsule powder containing hemp seed oil, 5-20 parts of xylo-oligosaccharide, 2-10 parts of Plantago ovata seed husk powder, 1-5 parts of konjak powder and 10-30 parts of lactitol by weight. Uniformly mixing solid microcapsule powder containing hemp seed oil, xylo-oligosaccharide, Plantago ovata seed shell powder, konjaku flour and lactitol; thus obtaining the composition with the function of relaxing bowel. The composition is prepared by compounding the fructus cannabis solid microcapsule powder and other materials with the same effect of relaxing bowel according to a specific proportion, achieves a better synergistic effect, and avoids resistance caused by long-term use of a single component.
Chinese invention patent application with application publication number CN 110269163A (application number 201810205567.0) discloses a bowel relaxing composition and a preparation method and application thereof, and a bowel relaxing beverage, which is mainly prepared from the following raw materials in parts by weight: 30-60 parts of rose, 0.5-10 parts of fruits and vegetables, 0.01-16 parts of Chinese herbal medicines, 30-60 parts of fructo-oligosaccharide, 1-5 parts of xylo-oligosaccharide, 0.1-2 parts of maltodextrin, 0-1 part of isomalto-oligosaccharide, 0-1 part of polydextrose, 0-1 part of bean peptide, 0-0.1 part of konjac flour, 0-0.01 part of vitamin E and 0-0.1 part of calcium lactate. The composition for relaxing bowel comprises various natural components for relaxing bowel, can promote intestinal peristalsis, soften excrement and improve intestinal microenvironment, can achieve the effect of improving constipation for a long time after being eaten for a proper period of time, and has no toxic or side effect.
The Chinese invention patent application with the application publication number of CN 109363187A (the application number of 201811076320.X) discloses a composition for regulating immunity, resisting fatigue and relaxing bowel and a preparation method thereof. The invention discloses a composition for regulating immunity, resisting fatigue and relaxing bowel and a preparation method thereof, wherein the composition comprises the following components in parts by weight: 800 parts of black bean protein 300, 50-250 parts of soybean protein, 50-250 parts of pea protein, 10-200 parts of walnut protein and 50-250 parts of fructo-oligosaccharide; the preparation method mainly adopts a wet spray granulation process and combines a dry mixing process if necessary. The composition provided by the invention achieves the effects of regulating immunity, resisting fatigue and relaxing bowel by reasonably combining and sharing various components, nourishing the five internal organs, tonifying the kidney and benefiting marrow, soothing the nerves and benefiting intelligence and regulating the balance of intestinal flora; the raw materials are safe to eat and easy to absorb, and experiments prove that the composition provided by the invention has excellent effects on immunity saving, fatigue resistance and bowel relaxing, and has very important practical popularization value.
Disclosure of Invention
The invention aims to provide a dendrobium compound preparation and a preparation method and application thereof, wherein each component of the dendrobium compound preparation plays its own role and forms a close and inseparable unified whole.
A dendrobium compound preparation comprises the following components by weight:
preferably, the dendrobium compound preparation consists of the following components in parts by weight:
preferably, in the dendrobium compound preparation, the mass ratio of the dendrobium devonianum to the fructus cannabis to the perilla fruit to the poria cocos to the Chinese date to the xylo-oligosaccharide is 8: 5: 4: 2: 4: 4. the above effective components have stronger synergistic effect in the mass ratio. The dendrobium compound preparation consists of the following components in parts by weight:
a preparation method of a dendrobium compound preparation comprises the following steps:
(1) wet-pulverizing and sieving Dendrobium devonianum to obtain powdery Dendrobium devonianum;
(2) respectively pulverizing fructus Cannabis, fructus Perillae, Poria, and fructus Jujubae by dry method, and sieving to obtain powdered fructus Cannabis, powdered fructus Perillae, powdered Poria, and powdered fructus Jujubae;
(3) putting the powdery dendrobium devoninum paxt obtained in the step (1), the powdery fructus cannabis, the powdery perilla fruit, the powdery poria cocos and the powdery Chinese date obtained in the step (2) into an extractor, adding water, and carrying out torrent wall breaking extraction to obtain an extracting solution;
(4) filtering the extracting solution obtained in the step (3) by plate-and-frame filter pressing to obtain filtrate;
(5) carrying out low-temperature concentration/drying integration technology on the filtrate obtained in the step (4), and crushing at low temperature to obtain dry powder;
(6) and (5) adding xylo-oligosaccharide and required auxiliary materials into the material obtained in the step (5), and preparing finished preparations with different forms according to the requirements.
In the step (1), the purple dendrobium is ground by a wet method and sieved by a 40-100 mesh sieve (preferably a 60 mesh sieve), so that powdery purple dendrobium is obtained. The method can effectively relieve the problem that the Dendrobium devonianum powder is easy to agglomerate when meeting water, and the powder Dendrobium devonianum below the size is suitable for extracting effective components.
In the step (2), the fructus cannabis, the perilla seeds, the poria cocos and the Chinese dates are respectively ground by a dry method and screened by a screen with 20-60 meshes (preferably a screen with 30 meshes) to obtain powdery fructus cannabis, powdery perilla seeds, powdery poria cocos and powdery Chinese dates. Materials below this size are suitable for extraction of active ingredients.
In the step (3), adding 40-60 times (preferably 50 times) of water to the mass of the materials in the step (1) and the step (2), namely 50 times of the mass of the raw materials of the powdery dendrobium devonianum in the step (1), the powdery fructus cannabis, the powdery perilla seed, the powdery poria cocos and the powdery Chinese date in the step (2), carrying out torrent wall breaking extraction at the temperature of 35-40 ℃ for 30-60 minutes. The extraction conditions are suitable for extracting effective components without damage.
And (4) filtering by adopting plate-and-frame filter pressing, wherein the filter cloth material comprises vinylon with a particle size of more than 100 meshes, and checking the transparency during filtering until the transparency meets the requirement. The method has high efficiency and is suitable for mass production.
The low-temperature concentration/drying integrated technology in the step (5) has the concentration temperature of 25-50 ℃, and the freeze drying technology can prevent the active ingredients from being damaged by high temperature.
And (3) adding xylo-oligosaccharide and different auxiliary materials according to requirements into the material obtained by extracting, filtering, concentrating and drying in the step (6), and preparing finished preparations with different forms according to requirements, so that different requirements can be met.
Although it is in the large intestine, constipation is closely related to the zang-fu organs, meridians, qi, blood, body fluids, spirit and emotion, and is a local manifestation of imbalance of yin and yang, zang-fu organs, qi, blood and emotion. For chronic habitual constipation or senile deficiency constipation caused by old and weak body and weakness after illness, deficient yin and blood, lusterless essence and blood, dry and astringent intestinal tract, difficulty in dredging and descending, and difficulty in running of dregs, dry stool and hard stool, and unsmooth discharge. The traditional Chinese medicine considers that aiming at constipation caused by intestine dryness due to deficiency of true yin and blood and body fluid, the traditional Chinese medicine is mainly used for nourishing yin, promoting the production of body fluid, tonifying qi and lubricating intestines.
The dendrobium is a traditional and rare traditional Chinese medicine in China, has the effects of nourishing yin and clearing heat, tonifying stomach and promoting fluid production, moistening lung and quenching thirst, moistening throat and improving eyesight, prolonging life and the like, has definite record on peculiar effects of dendrobium by medical works in past generations, is listed as a top-quality product in Shen nong herbal Jing, and is called sweet, mild and nontoxic. Mainly injures middle energizer, removes arthralgia, expels qi, tonifies five-treasure, asthenia, emaciation and strengthens yin. Taking the medicine for a long time to make intestines and stomach thick, and reducing weight and prolonging life; in compendium of materia medica, it records "Buyin Yijing" of Dendrobii. It can be taken for a long time to nourish intestines and stomach. Tonifying the interior, calming the stomach, strengthening the muscles, arresting will, eliminating convulsion, reducing weight and prolonging life. Dendrobium nobile is the key herb for nourishing yin, clearing heat, benefiting stomach and promoting fluid production, and is the best product for nourishing yin and tonifying deficiency. The dendrobium plants have various varieties, wherein the dendrobium devonianum (namely dendrobium devonianum) is a common variety in dendrobium medicinal materials, is also a dendrobium variety with published food standards, and has the property of medicine and food dual use.
The invention is based on the traditional Chinese medicine health preserving theory, and adopts the dendrobium devoninum paxt, fructus cannabis, perilla seeds, Chinese dates and poria cocos to form a new whole. Shi Pi Shi is sweet in flavor, slightly cold in nature, enters stomach and kidney meridians, excels in strengthening yin and replenishing vital essence, and thickens intestines and stomach, and is the monarch drug. Fructus cannabis is sweet in taste and neutral in nature, enters spleen, stomach and large intestine channels, has the effect of relaxing bowel, and is commonly used for treating blood deficiency, body fluid deficiency, intestinal dryness and constipation; perilla seed, fructus Perillae, being pungent and warm in flavor, enters lung meridian, lowers qi, relieves asthma, moistens intestines and relieves constipation, and the perilla seed are used as ministerial drugs together. Poria cocos is used as an adjuvant, has sweet and mild properties, has the effects of tonifying spleen and calming heart, has enough qi due to spleen health, and has enough qi to promote the production of body fluid of the dendrobium devonianum, and helps the generation and distribution of the body fluid of the dendrobium devonianum; the Chinese dates have the functions of tonifying middle-jiao and Qi, nourishing blood and tranquillizing mind and are guided by the rice. The dendrobium devoninum paxt, the fructus cannabis, the perilla seeds, the Chinese dates and the poria cocos are used together to play the roles of nourishing yin, promoting the secretion of saliva and body fluid, tonifying qi and lubricating intestines, so that the intestines are moistened by enough fluid and moved by enough qi. Can be used for treating constipation due to intestinal dryness caused by deficiency of true yin, blood deficiency and body fluid. The invention has the effects of promoting intestinal peristalsis, regulating immunity and the like according to modern pharmacology and health food functions.
According to the experimental structure during design, the dendrobium compound preparation has the functions of enhancing the immunity of RAW264.7 cells and promoting the increase of macrophages of model zebra fish, and the preparation has the function of regulating the immunity; the zebra fish bowel relaxing experiment shows that the invention has obvious intestinal peristalsis promoting effect; the mouse experiment shows that the composition has the function of regulating intestinal flora of constipation mice. Therefore, in the invention, the dendrobium devonianum has the effects of nourishing yin, enriching essence and thickening intestines and stomach, is matched with the fructus cannabis and the perilla fruit to nourish the intestines and relax the bowels, is matched with the poria cocos to invigorate the spleen and calm the heart, and is matched with the Chinese date to tonify the middle-jiao and qi, nourish the blood and calm the nerves, and the five components are combined to generate a synergistic effect, so that the dendrobium devonianum can consolidate the specimens and has good effects of regulating immunity, promoting intestinal peristalsis and regulating intestinal flora. The dendrobium compound preparation can be used for preparing functional foods and medicines with the functions of regulating immunity, promoting intestinal peristalsis and regulating intestinal flora.
Compared with the prior art, the method has the following advantages:
the invention is based on the traditional Chinese medicine health preserving theory, adopts the formula of the dendrobium devoninum paxt, the fructus cannabis, the perilla fruit, the Chinese date and the tuckahoe, each component plays its own role respectively, and forms a dense and inseparable unified whole. Shi Pi Shi is sweet in flavor, slightly cold in nature, enters stomach and kidney meridians, excels in strengthening yin and replenishing vital essence, and thickens intestines and stomach, and is the monarch drug. Fructus cannabis is sweet in taste and neutral in nature, enters spleen, stomach and large intestine channels, has the effect of relaxing bowel, and is commonly used for treating blood deficiency, body fluid deficiency, intestinal dryness and constipation; perilla seed, fructus Perillae, being pungent and warm in flavor, enters lung meridian, lowers qi, relieves asthma, moistens intestines and relieves constipation, and the perilla seed are used as ministerial drugs together. Poria cocos is used as an adjuvant, has sweet and mild properties, has the effects of tonifying spleen and calming heart, has enough qi due to spleen health, and has enough qi to promote the production of body fluid of the dendrobium devonianum, and helps the generation and distribution of the body fluid of the dendrobium devonianum; the Chinese dates have the functions of tonifying middle-jiao and Qi, nourishing blood and tranquillizing mind and are guided by the rice. The dendrobium devoninum paxt, the fructus cannabis, the perilla seeds, the Chinese dates and the poria cocos are used together to play the roles of nourishing yin, promoting the secretion of saliva and body fluid, tonifying qi and lubricating intestines, so that the intestines are moistened by enough fluid and moved by enough qi. Can be used for treating constipation due to intestinal dryness caused by deficiency of true yin, blood deficiency and body fluid. The invention has the functions of relaxing bowel and regulating immunity as described by referring to modern pharmacology and health food functions. Compared with the prior art, the traditional Chinese medicine composition is prepared according to the traditional Chinese medicine health preserving theory, is more rigorous and scientific in formula, gives consideration to the effects of relaxing bowel and regulating immunity, and has creative and obvious progress.
The components used in the invention are both medicinal and edible traditional Chinese medicines or food-grade ingredients, and can be directly used for producing common functional food without registering health-care food or medicines. Compared with the prior art, the use is more convenient and the conversion is easier.
The production process adopts a modern advanced production technology, the extraction process adopts a torrent wall breaking technology, the cell wall is completely broken under the action of water pressure, and the effective components in the plants are extracted at constant low temperature; the filtering process adopts plate-and-frame filter pressing, is suitable for filtering the crushed medicinal materials, has high efficiency and is convenient for industrial production; the concentration process is low-temperature concentration in a vacuum environment, so that the active ingredients are not damaged by high temperature; the drying process adopts freeze drying technology. The whole process does not need high-temperature treatment, so that the effective active ingredients can be effectively reserved, and the production process can save time and production cost. Extracting raw materials, filtering, concentrating, drying, adding adjuvants, and making into various dosage forms. The preparation method can more fully extract the effective components of the medicinal materials and retain the effective components, thereby achieving better effects of regulating immunity, promoting enterokinesia and regulating intestinal flora. The preparation method of the dendrobium compound preparation is advanced, is convenient for industrial production, and has wide application prospect. Compared with the prior art, the preparation method has high extraction yield, can better retain active substances without high-temperature treatment in the whole process, is suitable for industrial production, and has remarkable progress.
Drawings
FIG. 1 is a graph of the effect on the diversity of the intestinal flora.
Detailed Description
The present invention is further illustrated by the following examples, which should not be construed as limiting the scope of the invention.
Example 1
The dendrobium compound preparation comprises the following components in parts by weight:
80g of dendrobium devoninum paxt, 50g of fructus cannabis, 40g of perilla fruit, 20g of poria cocos, 40g of Chinese date and 40g of xylo-oligosaccharide.
The preparation is carried out according to the following specific mode:
(1) wet grinding the purple dendrobium, and sieving with a 60-mesh sieve to obtain powdery purple dendrobium;
(2) respectively crushing fructus Cannabis, fructus Perillae, Poria and fructus Jujubae by dry method, and sieving with 30 mesh sieve to obtain powdered fructus Cannabis, powdered fructus Perillae, powdered Poria and powdered fructus Jujubae;
(3) putting the powdery purple dendrobium officinale in the step (1), the powdery fructus cannabis, the powdery perilla seeds, the powdery poria cocos and the powdery Chinese date in the step (2) into an extractor, adding 50 times of water of the total mass of the raw materials, carrying out torrent wall breaking extraction at the temperature of 35-40 ℃, and extracting for 60 minutes to obtain an extracting solution;
(4) filtering the extracting solution obtained in the step (3) by plate-and-frame filter pressing (filter cloth material component vinylon, over 100 meshes) to obtain filtrate;
(5) concentrating the filtrate obtained in the step (4) at a low temperature of 40 ℃, freeze-drying, and crushing at a low temperature to obtain dry powder;
(6) and (4) adding xylo-oligosaccharide into the dry powder obtained in the step (5) according to the requirement, and granulating by a fluidized bed to prepare granules.
The dendrobium compound granules are taken by adults for 3g each time 2 times a day.
Example 2
The dendrobium compound preparation comprises the following components in parts by weight:
80g of dendrobium devoninum paxt, 50g of fructus cannabis, 40g of perilla fruit, 20g of poria cocos, 40g of Chinese date, 10g of xylo-oligosaccharide and a proper amount of magnesium stearate.
The preparation is carried out according to the following specific mode:
(1) wet grinding the purple dendrobium, and sieving with a 60-mesh sieve to obtain powdery purple dendrobium;
(2) respectively crushing fructus Cannabis, fructus Perillae, Poria and fructus Jujubae by dry method, and sieving with 30 mesh sieve to obtain powdered fructus Cannabis, powdered fructus Perillae, powdered Poria and powdered fructus Jujubae;
(3) putting the powdery purple dendrobium officinale in the step (1), the powdery fructus cannabis, the powdery perilla seeds, the powdery poria cocos and the powdery Chinese date in the step (2) into an extractor, adding 50 times of water of the total mass of the raw materials, carrying out torrent wall breaking extraction at the temperature of 35-40 ℃, and extracting for 30 minutes to obtain an extracting solution;
(4) filtering the extracting solution obtained in the step (3) by plate-and-frame filter pressing (filter cloth material component vinylon, over 100 meshes) to obtain filtrate;
(5) concentrating the filtrate obtained in the step (4) at low temperature of 30 ℃, freeze-drying, and crushing at low temperature to obtain dry powder;
(6) adding a proper amount of xylo-oligosaccharide into the dry powder obtained in the step (5) according to the requirement, and performing one-step granulation and particle finishing by a fluidized bed;
(7) and (4) uniformly mixing the granules prepared in the step (6) with a proper amount of magnesium stearate, and tabletting to obtain the dendrobium compound tablet.
The dendrobium compound tablet is taken by adults 2 times a day, 3 tablets each time.
Example 3
The dendrobium compound preparation comprises the following components in parts by weight:
60g of dendrobium devoninum paxt, 40g of fructus cannabis, 30g of perilla fruit, 10g of poria cocos, 30g of Chinese date, 10g of xylo-oligosaccharide and a proper amount of silica gel micropowder.
The preparation is carried out according to the following specific mode:
(1) wet grinding the purple dendrobium, and sieving with a 60-mesh sieve to obtain powdery purple dendrobium;
(2) respectively crushing fructus Cannabis, fructus Perillae, Poria and fructus Jujubae by dry method, and sieving with 30 mesh sieve to obtain powdered fructus Cannabis, powdered fructus Perillae, powdered Poria and powdered fructus Jujubae;
(3) putting the powdery purple dendrobium officinale in the step (1), the powdery fructus cannabis, the powdery perilla seeds, the powdery poria cocos and the powdery Chinese date in the step (2) into an extractor, adding 50 times of water of the total mass of the raw materials, carrying out torrent wall breaking extraction at the temperature of 35-40 ℃, and extracting for 30 minutes to obtain an extracting solution;
(4) filtering the extracting solution obtained in the step (3) by plate-and-frame filter pressing (filter cloth material component vinylon, over 100 meshes) to obtain filtrate;
(5) concentrating the filtrate obtained in the step (4) at a low temperature of 45 ℃, freeze-drying, and crushing at a low temperature to obtain dry powder;
(6) adding xylo-oligosaccharide into the dry powder obtained in the step (5) according to the requirement, and granulating and finishing the granules by a fluidized bed in one step;
(7) and (4) uniformly mixing the granules prepared in the step (6) with a proper amount of superfine silica gel powder, and filling into capsules to obtain the dendrobium devoninum paxt capsules.
The dendrobium compound capsule of the embodiment is taken by adults 2 times a day, 3 capsules each time.
Comparative example 1
Comparative example a dendrobium common preparation, consisting of the following components by weight:
40g of dendrobium devoninum paxt, 50g of fructus cannabis, 40g of perilla fruit, 20g of poria cocos, 40g of Chinese date and 40g of xylo-oligosaccharide.
The preparation method is the same as example 1.
Comparative example 2
Comparative example a dendrobium common preparation, consisting of the following components by weight:
80g of dendrobium devoninum paxt, 50g of fructus cannabis, 40g of perilla fruit, 20g of poria cocos, 40g of Chinese date and 40g of xylo-oligosaccharide.
The preparation is carried out according to the following specific mode:
(1) putting dendrobium devoninum paxt decoction pieces, fructus cannabis decoction pieces, perilla fruit decoction pieces, poria cocos decoction pieces and Chinese date decoction pieces into an extractor, adding water which is 50 times of the total mass of the raw materials, and extracting for 60 minutes at 95-100 ℃ to obtain extracting solution;
(2) filtering the extracting solution obtained in the step (1) by plate-and-frame filter pressing (filter cloth material component vinylon, over 100 meshes) to obtain filtrate;
(3) concentrating the filtrate obtained in the step (2) at 60-70 ℃, and performing spray drying to obtain dry powder;
(4) and (4) adding xylo-oligosaccharide into the dry powder obtained in the step (3) according to the requirement, and granulating by a fluidized bed to prepare granules.
The dendrobium compound granules are taken by adults for 3g each time 2 times a day.
Test of drug efficacy
Experiments prove that the invention has the functions of regulating immunity, promoting intestinal peristalsis and regulating intestinal flora.
Experiment for regulating immunity
1 Material
1.1 sample: the dendrobium compound preparation prepared in example 1; the dendrobium common preparation prepared in the comparative examples 1 and 2.
1.2 test cells: RAW264.7 cells were obtained from Shanghai bioscience institute of Chinese academy of sciences in DMEM supplemented with 10% heat-inactivated fetal bovine serum, 100U/mL penicillin and 100. mu.g/mL streptomycin at 37 ℃ with 5% CO2Culturing under the condition.
1.3 Experimental animals: the melanin allele mutant Albino strain zebrafish was bred in a natural pair mating breeding manner. 2 days (2dpf) after fertilization, feeding in 28 deg.C fish-farming water (water quality: 200mg of instant sea salt is added to per 1L of reverse osmosis water, conductivity is 480-510 μ S/cm, pH is 6.9-7.2, hardness is 53.7-71.6 mg/L CaCO)3) The license number for experimental animals is as follows: SYXK (Zhe) 2012-0171. The feeding management meets the requirements of international AAALAC certification.
1.4 Main reagents: RPMI1640 medium; DMEM high-glucose type; dual-antibody (penicillin and streptomycin) fetal bovine serum; 0.25% trypsin; tetrazolium blue (MTT); dimethyl sulfoxide (DMSO); an RNA extraction and purification kit; cDNA synthesis (reverse transcription) kit; a PCR kit; TNF-alpha, IL-6ELISA kit; vinorelbine; methyl cellulose; nile red.
1.5 Main instruments: CO 22An incubator; an ultra-clean bench; a microscope; a gradient PCR instrument; an ultraviolet-visible spectrophotometer; a low temperature bench centrifuge; an oscillator; an enzyme linked immunosorbent assay (ELISA) detector; a dissecting microscope; a camera connected to the microscope; 6, a hole plate; a precision electronic balance; a microinjection apparatus; a needle drawing instrument.
2 method
2.1 Effect on immune function of RAW264.7 cells
2.1.1 Effect on RAW264.7 cell proliferation
RAW264.7 cells (5X 10)5Cells) were plated in 96-well plates for 24 hours, and samples of different concentrations (50, 150, 450. mu.g/mL) were added to a final volume of 2mL at 37 ℃ with 5% CO2After incubation in an incubator for 18 hours 20. mu.L of LMTT (5g/L) was added at 37 ℃ with 5% CO2And (3) incubating the incubator for 4 hours, centrifuging, discarding the clear liquid, absorbing residual liquid by using filter paper, adding 100 mu LDMSO into each hole, slightly shaking for 10 minutes, detecting absorbance by using an enzyme-labeling instrument at 490nm wavelength, and calculating the relative cell proliferation rate which is the test group absorbance average value/blank group absorbance average value.
2.1.2 Effect on secretion of RAW264.7 cytokines
RAW264.7 cells (5X 10)5Individual cells) were inoculated in a 24-well flat-bottom plate for 24 hours, then each concentration sample was added to a final volume of 2mL, followed by 5% CO at 37 ℃2After 24 hours of culture in the incubator, cell supernatant is taken and the content of cytokines such as IL-6, TNF-alpha and the like is measured by an ELISA kit.
2.2 Effect on macrophage depletion of Zebra Fish
Randomly selecting 2 days (2dpf) after fertilization of the zebra fish of the melanin allelic mutant type Albino strain in a six-hole plate, treating 30 zebra fish in each hole (experimental group), and establishing a zebra fish macrophage reduction model by giving a dose of 0.25 ng/tail of vinorelbine through intravenous injection. The model zebra fish is respectively dissolved in water and given with samples with different dosages, and a normal control group (zebra fish treated by water for fish culture) and a model control group are arranged at the same time, and the volume of each hole (experimental group) is 3 mL. After the zebra fish is treated for 2 days, adding a neutral red solution to carry out vital body staining on the zebra fish, and counting the number of macrophages at the head of the zebra fish after the staining is finished; NR staining was also used to observe macrophage recruitment from zebrafish yolk and to evaluate statistically the macrophage formation-promoting effect of the samples.
3 results
3.1 immune function Effect on RAW264.7 cells
As can be seen from Table 1, in the concentration range of the experiment, the medium-high dose group of the sample can obviously stimulate the proliferation of RAW264.7 macrophage and obviously promote the release of cytokine IL-6, and the three dose groups can obviously promote the release of cytokine TNF-alpha, so that the sample has the function of regulating the immune function of RAW264.7 macrophage, and the effect is obviously better than that of the comparative example 1 group and the comparative example 2 group.
TABLE 1 Effect on immune function of RAW264.7 cells Experimental results
Note: p <0.05, P <0.01, P <0.001, compared to the blank control group.
3.2 Effect on macrophage depletion of Zebra Fish
Comparing the number of the macrophages at the head of the zebra fish in the model control group (39.8) with the number of the macrophages at the head of the normal control group (61.6) to ensure that the P is less than 0.001, so that the successful construction of the zebra fish macrophage reduction model is prompted; the numbers of the zebra fish head macrophages in a middle dose group (333.3 mug/mL) and a high dose group (1000 mug/mL) of the sample are 49.9 and 54.7 respectively, compared with a model control group, the promotion effects of the middle dose group P <0.01 and the high dose group P <0.001 on the zebra fish macrophage formation are 25.4% and 37.4% respectively, the result shows that the sample has obvious promotion effects on the macrophage formation, and the effect is obviously better than that of the comparative examples 1 and 2, and the result is shown in Table 2 in detail.
TABLE 2 evaluation of the results of the macrophage formation-promoting action
Note: p <0.01, P <0.001 compared to model controls.
Second, the experiment for promoting the intestinal peristalsis
1 Material
1.1 sample: the dendrobium compound preparation prepared in example 1; the dendrobium common preparation prepared in the comparative examples 1 and 2.
1.2 Experimental animals: wild type AB strain zebrafish, in a natural mated mating breeding mode. 5 days (5dpf) after fertilization, feeding in 28 deg.C fish-farming water (water quality: 200mg of instant sea salt is added to per 1L of reverse osmosis water, conductivity is 480-510 μ S/cm, pH is 6.9-7.2, hardness is 53.7-71.6 mg/L CaCO)3) The license number for experimental animals is as follows: SYXK (Zhe) 2012-0171. The feeding management meets the requirements of international AAALAC certification.
1.3 Main reagents: methyl cellulose; nile red; dimethyl sulfoxide; domperidone tablets.
1.4 Main instruments: a fluorescence microscope; a precision electronic balance; six orifice plates.
2 method
And randomly selecting wild type AB strain zebra fish 5 days (5dpf) after fertilization, dissolving in water, and giving Nile red to establish a zebra fish intestinal motility evaluation model. The test samples of the model zebra fish are subjected to water dissolving administration at concentrations of 111.1, 333.3 and 1000 mug/mL and the concentration of the positive drug domperidone is 30 mug/mL, meanwhile, a model control group is arranged, 30 zebra fish are treated in each hole (experimental group), and the volume of liquid medicine in each hole is 3 mL. After each group is treated for 24 hours, 10 zebra fishes are randomly selected from each group to be photographed under a fluorescence microscope and pictures are stored, NIS-Elements D3.10 advanced image processing software is used for image analysis and data collection, the intensity (S) of the zebra fish intestinal fluorescence signals is analyzed and counted, the promotion effect of the samples on the intestinal peristalsis is evaluated according to the statistical analysis result of the intensity of the intestinal fluorescence signals, and the statistical processing result is expressed by mean plus or minus SE. Statistical analysis using analysis of variance and Dunnett's T-test indicated significant differences with P < 0.05.
3 results
Zebra fish intestinal fluorescence signal intensity (6.60X 10) of positive control drug domperidone 30 mug/mL concentration group5Pixel) and model control group (12.76 × 10)5Pixel) comparison P<0.001, 48% of the intestinal peristalsis promoting effect, which indicates that the domperidone has obvious intestinal peristalsis promoting effect. The fluorescence signal intensities of the intestinal tracts of zebra fish in the groups with the sample concentrations of low, medium and high 111.1, 333.3 and 1000 mu g/mL are respectively 6.56 multiplied by 105、6.14×105And 6.86X 105Pixels, comparison with model control group P<0.001, the intestinal motility promoting effects were 49%, 52% and 46%, respectively. The samples are shown to have obvious intestinal peristalsis promoting effect under the experimental concentration condition, and the effect is obviously better than that of the groups of comparative examples 1 and 2. See table 3.
Table 3. influence experiment result on zebra fish intestinal fluorescence signal intensity
Note: p <0.001 compared to model control group.
Experiment for regulating intestinal flora
1 Material
1.1 sample: the dendrobium compound preparation prepared in the embodiment 1.
1.2 Experimental animals: clean grade ICR mice, male, weight 18 ~ 22 g. Provided by the experimental animal center in Zhejiang province, animal qualification number: SCXK (Zhe) 2019-. The license number for experimental animals is as follows: SYXK (Zhe) 2019-.
1.3 Main reagents: loperamide hydrochloride capsules;
pool DNA Kit (feces DNA extraction Kit);
fastpfu DNA Polymerase (ultra-high fidelity PCR enzyme); AxyPrepDNA gel recovery kit.
1.4 Main instruments: GeneAmp model 9700 PCR instrument; QuantiFluorTM-ST blue fluorescence quantification system; illumina MiSeq sequencer.
2 method
The method referred to in "technical Specification for health food inspection and evaluation (2003 edition)" is used for establishing a constipation model of mice by using loperamide hydrochloride. The experiment comprises a normal control group, a model control group, a low-dose test group (0.5g/kg), a medium-dose test group (1.0g/kg) and a high-dose test group (3.0g/kg), wherein each group contains 10 animals, the animals are administrated by gastric gavage for 10 days continuously for 1 time every day, and fresh feces are collected and are conveniently stored for later use at-80 ℃. The method adopts a 16S rRNA sequencing method to detect and analyze intestinal flora, obtains information such as diversity abundance of intestinal flora species through Alpha diversity analysis, and discusses the influence of the method on the intestinal flora of the constipation model mouse.
3 results
Constipation mice intestinal flora was detected by 16S rRNA gene sequencing method, Alpha diversity analysis showed: compared with a normal control, the number of the flora at the genus level of the model control group is obviously reduced by P <0.05, while the number of the flora at the genus level of each test group has no statistical difference, namely, the intestinal flora of the mouse at the model group is obviously reduced, and the test group can improve the reduction condition of the flora. The effect on the diversity of the intestinal flora is shown in FIG. 1.
Fourth, conclusion
Experimental results show that the dendrobium compound preparation can enhance the immune function of RAW264.7 cells, promote the increase of macrophages of model zebra fish, and has the function of regulating immunity; zebra fish experiments show that the invention has obvious intestinal peristalsis promoting effect, and the effect is obviously better than that of the groups of comparative examples 1 and 2; the mouse experiment shows that the composition has the function of regulating intestinal flora of constipation mice. Therefore, in the invention, the dendrobium devonianum has the effects of nourishing yin, enriching essence and thickening intestines and stomach, is matched with the fructus cannabis and the perilla fruit to nourish the intestines and relax the bowels, is matched with the poria cocos to invigorate the spleen and calm the heart, and is matched with the Chinese date to tonify the middle-jiao and qi, nourish the blood and calm the nerves, and the five components are combined to generate a synergistic effect, so that the dendrobium devonianum can consolidate the specimens and has better effects of regulating immunity, promoting intestinal peristalsis and regulating intestinal flora. The dendrobium compound preparation can be used for preparing functional foods and medicines with the functions of regulating immunity, promoting intestinal peristalsis and regulating intestinal flora.
Claims (10)
1. The dendrobe compound preparation is characterized by comprising the following components in parts by weight:
2. the dendrobium compound preparation of claim 1, which is characterized by comprising the following components in parts by weight:
3. the dendrobium compound preparation of claim 1, which is characterized by comprising the following components in parts by weight:
4. the preparation method of the dendrobium compound preparation according to any one of claims 1-3, which is characterized by comprising the following steps:
(1) wet-pulverizing and sieving Dendrobium devonianum to obtain powdery Dendrobium devonianum;
(2) respectively pulverizing fructus Cannabis, fructus Perillae, Poria, and fructus Jujubae by dry method, and sieving to obtain powdered fructus Cannabis, powdered fructus Perillae, powdered Poria, and powdered fructus Jujubae;
(3) putting the powdery dendrobium devoninum paxt obtained in the step (1), the powdery fructus cannabis, the powdery perilla fruit, the powdery poria cocos and the powdery Chinese date obtained in the step (2) into an extractor, adding water, and carrying out torrent wall breaking extraction to obtain an extracting solution;
(4) filtering the extracting solution obtained in the step (3) by plate-and-frame filter pressing to obtain filtrate;
(5) carrying out low-temperature concentration/drying integration technology on the filtrate obtained in the step (4), and crushing at low temperature to obtain dry powder;
(6) and (5) adding xylo-oligosaccharide and required auxiliary materials into the material obtained in the step (5), and preparing finished preparations with different forms according to the requirements.
5. The preparation method of the dendrobium compound preparation according to claim 4, wherein in the step (1), the dendrobium devonianum is subjected to wet grinding and sieved by a 40-100-mesh sieve to obtain powdery dendrobium devonianum.
6. The preparation method of the compound dendrobe preparation according to claim 4, wherein in the step (2), the fructus cannabis, the perilla fruit, the poria cocos and the Chinese date are respectively ground by a dry method and sieved by a 20-60-mesh sieve to obtain powdery fructus cannabis, powdery perilla fruit, powdery poria cocos and powdery Chinese date.
7. The preparation method of the dendrobium compound preparation according to claim 4, wherein in the step (3), the water adding amount is 40-60 times of the mass of the material, the torrent wall breaking extraction temperature is 35-40 ℃, and the extraction time is 30-60 minutes.
8. The preparation method of the compound dendrobium preparation according to claim 4, wherein in the step (4), the component of the plate-and-frame filter press cloth material, namely vinylon, is more than 100 meshes.
9. The method for preparing the compound preparation of dendrobium nobile lindl as claimed in claim 4, wherein in the step (5), the low-temperature concentration/drying integration technology comprises the following steps: the concentration temperature is 25-50 ℃, and the drying technology is a freeze drying technology.
10. The use of the compound preparation of dendrobium according to any one of claims 1 to 3 in the preparation of functional foods and medicines with the effects of regulating immunity, promoting intestinal peristalsis and regulating intestinal flora.
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