CN115281344A - Ganoderma lucidum polysaccharide composition for regulating female intestinal flora and preparation method thereof - Google Patents
Ganoderma lucidum polysaccharide composition for regulating female intestinal flora and preparation method thereof Download PDFInfo
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- CN115281344A CN115281344A CN202210935355.4A CN202210935355A CN115281344A CN 115281344 A CN115281344 A CN 115281344A CN 202210935355 A CN202210935355 A CN 202210935355A CN 115281344 A CN115281344 A CN 115281344A
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- ganoderma lucidum
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Abstract
The invention provides a ganoderma lucidum polysaccharide composition for regulating female intestinal flora and a preparation method thereof, wherein the ganoderma lucidum polysaccharide composition comprises the following raw materials of ganoderma lucidum polysaccharide, procyanidine of sea buckthorn seeds, mulberry leaf extract, houttuynia cordata extract, raspberry fruit powder, red date extract and chia seed powder, and the raw materials are weighed according to the weight part ratio and are uniformly mixed to obtain the ganoderma lucidum polysaccharide composition; wherein the ganoderan consists of 7 monosaccharides including fucose, galactose, glucose, xylose, mannose, glucuronic acid and glucosamine. The ganoderma lucidum polysaccharide composition furthest exerts the effect of regulating female intestinal flora by ganoderma lucidum polysaccharide, and exerts the complementary optimization of each raw material on the effects, aroma and taste by compounding each raw material.
Description
Technical Field
The invention belongs to the technical field of functional foods, and particularly relates to a ganoderan composition for regulating female intestinal flora and a preparation method thereof.
Background
The intestinal tract is the most important nutrient absorption organ of human body, participates in various physiological activities such as food digestion, nutrient absorption, energy supply, essential vitamin generation, immunoregulation, gastrointestinal tract homeostasis and the like, and is closely related to the occurrence and development of various metabolic diseases and inflammations. The intestinal tract is also the largest immune organ of the human body, intestinal epithelial cells also form a mucosal immune barrier besides the digestive metabolic function, and more than 70 percent of immune cells of the human body exist in the intestinal mucosa. A huge number of microbial communities exist in the intestinal tract, are the 'second set of genome' of a human body, and have an important role in establishing an intestinal mucosa immune system, and after the intestinal flora is fixedly planted and perfected, a mycoderm with an important physiological function is formed on the intestinal mucosa. Anaerobic bacteria such as bifidobacterium, lactobacillus and the like are combined with specific receptors on the surface of the intestinal mucosa epithelium and are tightly adhered to the intestinal mucosa epithelium to form a biomembrane structure and a biological barrier with relatively fixed components, so that the intestinal flora colonization resistance is formed, and the invasion of exogenous substances to organisms can be effectively resisted. The intestinal flora and the intestinal mucosa immune system are co-evolved to jointly maintain the intestinal homeostasis. The intestinal flora is closely related to the change of the barrier function of the intestinal tract, and plays an important role in promoting the development of the intestinal mucosa immune system, regulating and controlling the intestinal mucosa immunity, resisting infection and the like. The normal flora plays an important role in regulating the development of various lymphocyte subsets in the intestinal tract, and the interaction of the nutrition and the metabolism of the intestinal tract obviously influences the health and the immune condition of the organism.
The selective administration of prebiotics, probiotics and synbiotics is an important way to maintain the balance of the intestinal flora. Prebiotics are substances that are not utilized by harmful pathogenic bacteria in the intestinal tract, but are utilized in large quantities by probiotics, thereby enhancing the competitive advantage of the probiotics. The concept was first proposed by Gibson in 1995, and then as related research advances and applications expand, gibson, food and agriculture organization of the united nations, etc. revised prebiotic definitions many times. The 2016 international association of probiotics and prebiotics science (ISAPP) expert group, based on the latest scientific and clinical evidence, updated the definition of prebiotics (a substrate that can be selectively utilized by microorganisms and confers health benefits to the host), and products that clearly indicate the ability to be selectively utilized by host microorganisms to produce health benefits are all called prebiotics, meaning that research in this field is entering a new stage of development. At present, the more mature prebiotics are mainly various oligosaccharides including fructo-oligosaccharide, galacto-oligosaccharide, inulin, breast milk oligosaccharide and the like, and certain polysaccharides are regarded as high-quality sources of novel prebiotics due to the comprehensive effects of promoting the proliferation of probiotics, improving the intestinal health of organisms, improving the immunity, preventing tumors and the like, but the research of the novel prebiotics is just started, the interaction research between the prebiotics and the intestinal flora is not deep enough, and the systematicness is not strong.
The source and structure of the polysaccharide have a great influence on the efficacy of the polysaccharide, and the chain length, the type of glycosidic bond, the connection mode and the molecular mass all have influence on the overall efficacy and the regulation effect of intestinal flora. At present, a complete system is not formed in the screening and efficacy evaluation of the probiotics polysaccharide, the product variety of the comprehensive efficacy polysaccharide can not meet the requirement, and the screening of the novel probiotics polysaccharide is in the vogue. Wherein, the ganoderma lucidum is an important traditional Chinese medicinal material in Zhejiang, the content of polysaccharide is high, and the ganoderma lucidum polysaccharide is an important bioactive substance, and the influence of the ganoderma lucidum polysaccharide on the human intestinal flora is not clear.
Disclosure of Invention
Aiming at the problems in the prior art, the invention aims to provide a ganoderan composition for regulating female intestinal flora and a preparation method thereof, which furthest exert the effect of regulating female intestinal flora by ganoderan and exert the complementary optimization of the effects, aroma and taste of all raw materials by compounding all raw materials.
The purpose of the invention is realized by the following technical scheme:
a ganoderan composition for regulating female intestinal flora comprises ganoderan, fructus Hippophae procyanidine, a mulberry leaf extract, a houttuynia cordata extract, raspberry fruit powder, a red date extract and chia seed powder, and the weight parts of the raw materials are as follows:
weighing the raw materials according to the weight part ratio, and uniformly mixing to obtain the ganoderma lucidum polysaccharide composition.
Further, the method for purifying the ganoderma lucidum polysaccharide comprises the following steps: drying Ganoderma lucidum fruiting body at 40-70 ℃, sieving with a 50-mesh sieve after crushing, soaking for 24h according to 8mL volume times of 95% ethanol of 1g dry powder, removing lipids and small molecular substances, drying filter residues under the above conditions, uniformly mixing according to the proportion of 10-30mL water of 1g powder, carrying out 500w ultrasonic treatment for 30min, carrying out 40-90 ℃ water bath treatment for 60-120min, centrifuging to obtain supernatant, precipitating for 4-8h with 5 volume times of 95% ethanol, centrifuging to collect precipitate, dissolving in deionized water again according to the concentration of 5mg/mL, dialyzing for 72h with a 5000Da bag, passing through a sephadex column to obtain a single component, carrying out vacuum concentration on the obtained eluate, and carrying out freeze drying to obtain purified Ganoderma lucidum polysaccharide.
Further, the ganoderma lucidum polysaccharide is composed of 7 monosaccharides, namely fucose, galactose, glucose, xylose, mannose, glucuronic acid and glucosamine.
Further, the preparation method of the seabuckthorn seed procyanidin comprises the following steps: taking the seabuckthorn seed meal after extracting the seabuckthorn seed oil as a raw material, carrying out flash extraction by using an ethanol solution at a low temperature, and filtering to obtain an extracting solution; recovering ethanol from the extract at low temperature under negative pressure by using a heat pump device, and concentrating the extract to obtain an extract; and (3) the extract is subjected to a belt drying device at a low temperature and under a negative pressure condition to obtain powdery seabuckthorn seed procyanidine.
Further, the preparation method of the mulberry leaf extract comprises the following steps: cutting fresh mulberry leaves into strips, soaking the strips by using an acetic acid solution, filtering, collecting filtrate, centrifuging the obtained filtrate, collecting supernatant, concentrating and drying the supernatant to solid so as to obtain the mulberry leaf extract.
Further, the preparation method of the houttuynia cordata extract comprises the following steps: cutting fresh houttuynia cordata to obtain houttuynia cordata powder, decocting the powder with water under the condition of microwave and ultrasonic wave combined auxiliary dissolution, filtering to obtain houttuynia cordata extract, concentrating the extract under reduced pressure under the vacuum condition to obtain thick paste, and preparing the thick paste into the houttuynia cordata extract by a spray drying machine.
Further, the preparation method of the raspberry fruit powder comprises the following steps: cleaning fresh raspberry fruits without pedicels, crushing to prepare raspberry fruit pulp, adding an enzyme preparation, and performing enzymolysis at room temperature for 1-4h; passing the raspberry pulp after enzymolysis through a homogenizer for 2-3 times to obtain homogenized raspberry pulp; and (3) passing the raspberry slurry through a high-pressure homogenizer to obtain the raspberry slurry after high-pressure homogenization, and performing low-temperature spray drying on the raspberry slurry to obtain raspberry fruit powder.
Further, the preparation method of the red date extract comprises the following steps: taking dry red dates as raw materials, washing, soaking, crushing into red date pieces, adding an enzyme preparation, carrying out enzymolysis for 1-4h at 40-50 ℃, filtering to obtain supernatant, and carrying out vacuum concentration on the supernatant to obtain semi-solid red date paste; and (3) carrying out vacuum drying, crushing and sieving treatment on the semi-solid red date paste to obtain the red date extract.
Further, the preparation method of the chia seed powder comprises the following steps: removing impurities from seeds of chia sold in the market, cleaning, airing, grinding into powder, and sieving with a 200-mesh sieve to obtain the chia seed powder.
Another aspect of the invention:
the application of the ganoderan composition is used for preparing foods, medicines or health-care products for preventing and/or regulating female intestinal flora disorder.
Further, the food, the medicine or the health-care product comprises other acceptable auxiliary materials in the food, the medicine or the health-care product, and the auxiliary materials are one of powder, tablets, granules, capsules or pills.
Another aspect of the invention:
the application of the mixed components of chia seed powder, raspberry fruit powder and red date extract in improving bitterness and fishy taste and covering fishy smell is as follows:
5-10 parts of raspberry fruit powder
5-10 parts of red date extract
3-6 parts of chia seed powder.
Compared with the prior art, the invention has the beneficial effects that:
1. according to the invention, the influence of the ganoderan composition on the composition of intestinal flora and important metabolites thereof of healthy adults is researched, so that the ganoderan composition can reduce the abundance of proteobacteria and increase the relative abundance of bacteroides; and analysis on a subordinate level shows that the addition of the ganoderan composition can reduce the relative abundance of escherichia coli-shigella regardless of men and women and regardless of age groups, and particularly the reduction range of women is greater than that of men; after the ganoderma lucidum polysaccharide composition is added, the abundance of bifidobacteria of women shows an increasing trend, the original abundance of middle-aged women is lower, and the increase amplitude is higher; therefore, the ganoderan composition is suitable for preparing foods, medicines or health-care products for preventing and/or regulating the female intestinal flora disorder;
2. according to the invention, the ganoderma lucidum polysaccharide composition is prepared by selecting several raw materials of ganoderma lucidum polysaccharide, sea-buckthorn seed procyanidine, mulberry leaf extract, houttuynia cordata extract, raspberry fruit powder, red date extract and chia seed powder, and the complementary optimization of the raw materials in terms of efficacy, fragrance and taste is exerted; the mulberry leaf extract and the houttuynia cordata extract have good health-care effects of dispelling wind and heat, clearing away the lung-heat, moistening dryness, resisting bacteria and viruses and repairing intestinal mucosa, but have bitter and astringent tastes and fishy smells, the application of the mulberry leaf extract and the houttuynia cordata extract in foods or health-care products is greatly influenced, the raspberry fruit powder and the red date extract have special sour and sweet tastes and strong fragrance, the chia seed powder is smooth in taste, rich in dietary fibers and capable of moistening intestines and expelling toxins, the bitter and astringent tastes and the fishy smells of the mulberry leaf extract and the houttuynia cordata extract can be wrapped and passivated, the bitter and fishy smells of the mulberry leaf extract and the houttuynia cordata extract are well improved after mixing, the fishy smells are covered, and meanwhile, the sea buckthorn seed procyanidin and the raspberry fruit powder are rich in phytocyanidin, and the effects of resisting oxidation and cleaning blood are added to the composite product;
3. in the ganoderma lucidum polysaccharide composition, the mulberry leaf extract and the houttuynia cordata extract are added, so that the effects of diminishing inflammation and repairing intestinal mucosa are exerted, and the beneficial effects of the ganoderma lucidum polysaccharide in regulating intestinal flora disorder and repairing intestinal cell damage are assisted and enhanced;
4. from the aspect of food supplement of traditional Chinese medicine, the red date extract and chia seed powder in the ganoderma lucidum polysaccharide composition belong to warm food, the addition of the red date extract and chia seed powder can neutralize the cold property of the mulberry leaf extract and the houttuynia cordata extract, and the composition is smooth in nature after the raw materials are combined, and is suitable for women to drink all the year round; meanwhile, the chia seed powder is rich in dietary fiber, vitamins and polyunsaturated fatty acid, is matched with natural antioxidant seabuckthorn seed procyanidin, can obviously reduce the generation of prostaglandin PGF2 alpha and prostaglandin LT4 and free radicals which cause female dysmenorrheal, is suitable for being eaten by women during or before menstrual period, and has the function of relieving dysmenorrheal.
Drawings
The invention is further illustrated by the following examples in conjunction with the accompanying drawings:
FIG. 1 is a graph of total sugar standards;
FIG. 2 is a diagram of the molecular weight distribution of ganoderan;
FIG. 3 is a HPLC chart of the standard, wherein the left is other monosaccharides and the right is glucosamine;
FIG. 4 is a graph showing the results of chao1 and shannon index;
FIG. 5 is a graph of PCA for different population combinations;
FIG. 6 is a graph of the results of the effect of ganoderan compositions on gut flora composition (phylum level);
FIG. 7 is a graph showing the effect of ganoderan composition on the composition of intestinal flora (genus level);
FIG. 8 is a graph showing the effect of ganoderan composition on the production of short chain fatty acids by intestinal flora;
FIG. 9 is a graph showing the effect of ganoderan composition on gas production by gut flora.
Detailed Description
Example 1 ganoderan compositions and preparation thereof
The embodiment provides a ganoderan composition for regulating female intestinal flora, which consists of ganoderan, seabuckthorn seed procyanidine, a mulberry leaf extract, a houttuynia cordata extract, raspberry fruit powder, a red date extract and chia seed powder, wherein the ganoderan composition comprises the following raw materials in parts by weight:
weighing the raw materials according to the weight part ratio, and uniformly mixing to obtain the ganoderma lucidum polysaccharide composition which is mixed powder. When the ganoderma lucidum polysaccharide composition is eaten, a proper amount of ganoderma lucidum polysaccharide composition is taken, warm water is added, and the ganoderma lucidum polysaccharide composition is stirred and dissolved, so that the ganoderma lucidum polysaccharide composition can be eaten.
The method for purifying the ganoderma lucidum polysaccharide comprises the following steps: drying Ganoderma lucidum fruiting body at 40-70 deg.C, pulverizing, sieving with 50 mesh sieve, soaking in 8mL volume times of 95% ethanol for 24h according to 1g dry powder, removing lipid and small molecular substances, drying the filter residue under the above conditions, mixing uniformly according to the proportion of 10-30mL water of 1g powder, performing 500w ultrasonic treatment for 30min, performing water bath treatment at 40-90 deg.C for 60-120min, centrifuging to obtain supernatant, precipitating with 5 volume times of 95% ethanol for 4-8h, centrifuging to collect precipitate, dissolving in deionized water again according to 5mg/mL concentration, dialyzing with 5000Da bag for 72h, passing through Sephadex column to obtain single component, vacuum concentrating the obtained eluate, and freeze drying to obtain purified Ganoderma lucidum polysaccharide.
The total sugar and the reducing sugar are measured by two methods, namely an absorbance method and an enzyme-labeling method, and an intermediate value is taken. The absorbance method and the enzyme-linked immunosorbent assay are both used for determining the purity of the ganoderma lucidum polysaccharide, and the purity is 75%. The total sugar standard curve is shown in figure 1. The molecular weight of ganoderan was determined by HPLC method, and is shown in FIG. 2 and Table 1.
TABLE 1 molecular weight information of ganoderan
As can be seen from FIG. 2 and Table 1, the weight average molecular weight of ganoderan is lower, 21043. The monosaccharide composition is determined by ion chromatography, 11 monosaccharides of fucose, arabinose, rhamnose, galactose, glucose, xylose, fructose, mannose, galacturonic acid, glucuronic acid and glucosamine are used as standard control, and HPLC chart and information are shown in FIG. 3 and Table 2.
TABLE 2 Standard substance types and related information (concentration 5 mg/L)
TABLE 3 monosaccharide composition of ganoderan
As can be seen from table 3, the ganoderan has a relatively complex composition, and consists of 7 monosaccharides, i.e., fucose, galactose, glucose, xylose, mannose, glucuronic acid, and glucosamine, wherein the glucose content is the highest and is 67%, the glucuronic acid content is lower and is about 3.4%, and the content of the remaining 5 monosaccharides is not much different and is between 5 and 7%, which increases many difficulties for the analysis of the fine structure.
The preparation method of the seabuckthorn seed procyanidin comprises the following steps: taking semen Hippophae dregs after extracting oleum Hippophae as raw material, extracting with 60% ethanol solution at low temperature of 15 deg.C for 2-3 times with flash extraction for 10min each time, mixing 2-3 times of extractive solutions, and filtering with 300 mesh filter cloth to obtain extractive solution; recovering ethanol from the filtered extractive solution at low temperature of 15 deg.C and negative pressure of-0.098 MPa by heat pump device, and concentrating the extractive solution to obtain extract; and (3) the extract is processed by a belt type drying device at the low temperature of 15 ℃ and under the negative pressure of-0.098 MPa to obtain the powdery seabuckthorn seed procyanidine.
The preparation method of the mulberry leaf extract comprises the following steps: cutting fresh mulberry leaves into strips with the width of 1cm, soaking for 1-3h at the temperature of 45 ℃ by using an acetic acid solution with the concentration of 0.05M, filtering by using gauze, collecting filtrate, centrifuging the obtained filtrate for 10-20min at 3000r/M, collecting supernatant, concentrating and drying to solid, thereby obtaining the mulberry leaf extract.
The preparation method of the houttuynia cordata extract comprises the following steps: cutting fresh houttuynia cordata to obtain houttuynia cordata powder, decocting the powder with water for 2 to 4 hours under the condition of microwave (power of 400 to 500W) and ultrasonic (power of 600 to 750W) combined auxiliary dissolution, filtering to obtain houttuynia cordata extract, concentrating the extract under reduced pressure under the vacuum condition (vacuum degree of 0.06MPa to 0.08 MPa) to form thick paste, and preparing the houttuynia cordata extract from the thick paste through a spray drying machine.
The preparation method of the raspberry fruit powder comprises the following steps: cleaning fresh raspberry fruits without pedicels, crushing to prepare raspberry fruit pulp, adding an enzyme preparation, and performing enzymolysis at room temperature for 1-4h; passing the enzyme-hydrolyzed raspberry pulp through a homogenizer for 2-3 times to obtain homogenized raspberry pulp; and (4) passing the raspberry slurry through a high-pressure homogenizer to obtain the raspberry slurry after high-pressure homogenization, and carrying out low-temperature spray drying on the raspberry slurry to obtain the raspberry fruit powder.
The raspberry has full fruits and ruddy color, can be picked directly for eating, tastes sour and sweet, and has higher nutritional value. The raspberry contains various functional components such as flavonoid compounds, ellagic acid, anthocyanin, salicylic acid, raspberry ketone and the like besides abundant amino acids, minerals and vitamins; has various biological effects of removing chloasma, improving dysmnesia, treating osteoporosis, treating gynecological diseases and the like, and has the reputation of life fruit. The raspberry fruit powder obtained after enzymolysis has special sour and sweet taste and strong fragrance.
The preparation method of the red date extract comprises the following steps: taking dry red dates as raw materials, washing, soaking, crushing into red date pieces, adding an enzyme preparation, carrying out enzymolysis for 1-4h at 40-50 ℃, filtering to obtain supernatant, and carrying out vacuum concentration on the supernatant to obtain semi-solid red date paste; and (3) carrying out vacuum drying, crushing and sieving treatment on the semi-solid red date paste to obtain the red date extract.
In the preparation of the raspberry fruit powder and the red date extract, the enzyme preparations are mixed enzyme preparations of cellulase and pectinase.
The preparation method of the chia seed powder comprises the following steps: removing impurities from seeds of chia sold in the market, cleaning, airing, grinding into powder, and sieving with a 200-mesh sieve to obtain the chia seed powder.
Comparative examples 1 to 5
The comparative example provides a ganoderan composition, and the components and parts by weight of the raw materials are shown in the following table 4:
TABLE 4 COMPARATIVE EXAMPLES 1-5 TABLE FOR COMPARATIVE EXAMPLES OF LINGZHITAN POLYSACCHARIDE COMPOSITIONS
Raw materials | Comparative example 1 | Comparative example 2 | Comparative example 3 | Comparative example 4 | Comparative example 5 |
Ganoderma lucidum polysaccharide | 45 | 45 | 45 | 45 | 45 |
Mulberry LeavesExtract of plant | 6 | 6 | 6 | 6 | 6 |
Houttuynia cordata extract | 3.5 | 3.5 | 3.5 | 3.5 | 3.5 |
Sea-buckthorn seed procyanidine | 7 | 7 | 7 | 7 | 7 |
Chia seed powder | / | 24 | / | / | 4.5 |
Raspberry fruit powder | / | / | 24 | / | 19.5 |
Red date extract | / | / | / | 24 | / |
Weighing the raw materials according to the weight part ratio in the table above, and uniformly mixing to obtain the ganoderma lucidum polysaccharide composition which is mixed powder. When the ganoderma lucidum polysaccharide composition is eaten, a proper amount of ganoderma lucidum polysaccharide composition is taken, warm water is added, and the ganoderma lucidum polysaccharide composition is stirred and dissolved, so that the ganoderma lucidum polysaccharide composition can be eaten.
The preparation of each starting material was as described in example 1.
The trained personnel perform sensory evaluation on the ganoderan composition described in example 1 and comparative examples 1-5, wherein the fishy smell evaluation score of comparative example 1 (without chia seed powder, raspberry fruit powder and red date extract added) is set to 10.
The sensory evaluation results of the examples and comparative examples are shown in table 5. The results show that the three mixed components of chia seed powder, raspberry fruit powder and red date extract have better regulating effect on the fishy smell than the combination of any one or two of the components. The raspberry fruit powder and the red date extract have special sour and sweet taste and strong fragrance, the chia seed powder is soft and smooth in taste, is rich in dietary fiber, has the effects of moistening intestines and expelling toxin, and can wrap and inactivate bad taste including bitter and astringent taste and fishy smell.
TABLE 5 sensory evaluation results of ganoderan compositions
Numbering | Comparative example 1 | Comparative example 2 | Comparative example 3 | Comparative example 4 | Comparative example 5 | Example 1 |
Sensory evaluation score | 10 | 8.0 | 6.5 | 6.0 | 6.5 | 2.0 |
Example 2 Regulation of the intestinal flora by a ganoderan composition
32 healthy adults were divided into four groups, namely 20-30 years old, named YM (yang male), YF (yang male), 40-50 years old, named MM (middle male), MF (middle male), GC (control group) without ganoderan composition, LZ (lingzhi) with ganoderan composition (5 g/L), and GC-YM, LZ-YF, GC-MM, and LZ-MF.
1. Influence on diversity
The Chao1 index is an index for measuring the abundance of a species, and is independent of abundance and uniformity, but is sensitive to rare species. The Shannon index is related to the number and uniformity of individual distribution (uniformity or evenness) among the populations, and the more uniform the individual distribution among the populations, the greater the Shannon index. The diversity index is maximal if each individual belongs to a different species; if each individual belongs to the same species, its diversity index is minimal. The larger the shannon index, the better the sample homogeneity is reflected. As can be seen from FIG. 4, the ganoderan composition has no significant effect on the abundance and diversity of human intestinal flora; the flora abundance of middle-aged men is increased; the flora diversity of women is slightly reduced.
PCA (Principal Components Analysis) is used to characterize beta-diversity, a projection technique that refers to mapping data in a high-dimensional space to a low-dimensional space. For example, a sphere in a three-dimensional space is projected in the coordinate axis direction to form a circle, and in the projection process of changing the sphere into the circle, part of the properties of the original object are lost and retained, so that the method is helpful for reducing multi-dimensional information into two-dimensional information and visually understanding data.
As can be seen from fig. 5, the ganoderan composition has no significant effect on the composition of male and female flora, i.e. beta-diversity, regardless of the young and middle aged groups, indicating that it has no significant effect on the stability of the intestinal flora of healthy people and has potential for application as a daily health food.
2. Influence on changes in intestinal flora
The human intestinal flora mainly comprises actinomycetomycota, bacteroidetes, firmicutes, proteobacteria and the like, wherein the bacteroidetes generally refers to bacteria which grow in human intestinal tracts and stomachs and play a positive role in human health, the abundance of the bacteroides is generally increased to be beneficial to the human health, some potential probiotics, namely the bacteroides parapsilosis also belong to the phylum at present, and a large amount of bacteroides in fecal intestinal flora of some pathogenic bacteria infected patients is reduced or even disappears. The phylum firmicutes includes Bacillus, enterococcus, lactobacillus, lactococcus, leuconostoc, acetobacter, etc., including many probiotic strains; furthermore, the higher the ratio of firmicutes to bacteroides, the more efficient the absorption of nutrients in food, and conversely the potential for weight loss. Proteobacteria comprise various pathogenic bacteria, such as escherichia coli, salmonella, cholera navy, helicobacter pylori and the like, the microecological imbalance process during metabolic disorder is accompanied by the increase of the Proteobacteria, and the dominant Proteobacteria also exist in the intestinal tracts of children with malnutrition, so that immune imbalance and inflammation are caused, and researchers think that the Proteobacteria is a potential identification standard for occurrence of microecological imbalance and diseases.
As can be seen from fig. 6, in the scope of the present experiment, from the phylum level, after the ganoderan composition described in example 1 is added, the relative abundance of bacteroides is significantly increased in the case of both men and women, and the abundance of proteobacteria is significantly decreased. The relative abundance of male firmicutes is not large in change range, the relative abundance of the firmicutes of young women is slightly increased, the relative abundance of the firmicutes of middle-aged women is higher, and the abundance is slightly reduced after the ganoderma lucidum polysaccharide composition is added. The relative abundance of actinomycete phyla in males has a descending trend; the relative abundance of actinomycete phyla in women has little change, and the middle-aged group has slightly increased.
As can be seen from fig. 7, analysis at a subordinate level, addition of the ganoderan composition described in example 1 reduced the relative abundance of escherichia coli-shigella regardless of male and female, but the reduction was greater in female than male; the relative abundance of the four bacteroidetes is obviously increased, and the increase range of the middle-aged people is larger than that of the young people; the abundance of bifidobacteria in young men is increased, the original relative abundance of bifidobacteria in middle-aged men is higher, the relative abundance of bifidobacteria in women is increased when the ganoderan described in example 1 is added, and the original abundance of bifidobacteria in middle-aged women is lower and the increase amplitude is higher; four groups of Prevotella, coorales and enterococcus showed a remarkable increase. The ganoderma lucidum polysaccharide of example 1 can regulate the intestinal flora structure of human body and develop towards a healthier direction.
3. Influence on short-chain fatty acid production by intestinal flora
This example separately investigated the effect of the ganoderan composition described in example 1 on the metabolism of the gut flora to produce acetic, propionic, butyric, isobutyric, valeric, isovaleric 6 short chain fatty acids and total acids. It was found that the ganoderan composition of example 1 significantly promotes the production of acetic acid in both male and female groups, both young and middle age groups; the trend of propionic acid and butyric acid is also the same, and the trend is obviously increased; valeric acid changes were insignificant and total short chain fatty acid content was significantly increased. Therefore, the ganoderan composition in example 1 can obviously increase the content of short-chain fatty acids in the intestinal tract, and has important significance in reducing the pH value of the intestinal tract, improving the composition of the intestinal flora and maintaining a healthy environment.
4. Influence on gas production of intestinal flora
This example investigated the metabolism of intestinal flora by CH produced by the ganoderan composition described in example 1 4 、NH 3 、H 2 S、H 2 、CO 2 The influence of five gases. It was found that the ganoderan composition described in example 1 was able to significantly reduce CH in 4 groups 4 Yield, H of young male group 2 S content also tends to decrease, excessive CH 4 And H 2 S has adverse effects on the composition of intestinal epithelial cells and flora; NH 3 The content of (A) is not changed greatly and has an ascending trend; h 2 With CO 2 The two neutral gas contents are obviously increased, and probably are related to the improvement of the expected flora composition, and the influence and the principle of the two neutral gas contents need to be further studied deeply.
And (4) conclusion:
the ganoderma lucidum polysaccharide is further purified in the embodiment 1 of the invention, and the purity reaches 75%; the average molecular weight of the polysaccharide is 21KD, the composition of the polysaccharide is relatively complex, the polysaccharide consists of 7 monosaccharides including fucose, galactose, glucose, xylose, mannose, glucuronic acid and glucosamine, wherein the glucose content is the highest and accounts for 67%, the glucuronic acid content is lower and accounts for about 3.4%, and the content of the other 5 monosaccharides is not greatly different and is between 5 and 7%.
Example 2 the effect of the composition of ganoderan according to example 1 on the composition of the intestinal flora and the composition of important metabolites thereof (8 people are group 1, and are classified according to age and gender, and are respectively GC-YM, LZ-YF, GC-MM and LZ-MF groups) of 32 healthy adults was studied, and from the phylum level, the addition of the ganoderan composition according to example 1 can reduce the abundance of proteobacteria and increase the relative abundance of Bacteroides, with slightly different changes in male and female; analysis on the subordinate level, addition of the ganoderan composition described in example 1 was able to reduce the relative abundance of escherichia coli-shigella, both in men and in women, but the reduction was greater in women than in men; the relative abundance of 4 groups of bacteroidetes is obviously increased, and the increase range of middle-aged people is larger than that of young people; the abundance of bifidobacteria in young men is increased, the original relative abundance of bifidobacteria in middle-aged men is higher, the relative abundance of bifidobacteria is reduced after the ganoderan composition in example 1 is added, the abundance of bifidobacteria in women tends to increase, and the original abundance of bifidobacteria in middle-aged women is lower and has higher increase amplitude; four groups of Prevotella, coula and enterococcus showed a remarkable increase.
The ganoderan composition in example 1 has a relatively obvious effect of promoting the generation of important short-chain fatty acids such as acetic acid, propionic acid and butyric acid, and is suitable for male or female, young or middle-aged people; the total short chain fatty acid content is obviously increased. The ganoderan composition described in example 1 can significantly reduce CH 4 、H 2 Production of harmful gases such as S, but to H 2 With CO 2 The increase in the content of the two neutral gases has a promoting effect, which may be related to the improvement of the flora composition thereof, for reasons to be further investigated.
Finally, it should be noted that the above only illustrates the technical solution of the present invention, but not limited thereto, and although the present invention has been described in detail with reference to the preferred arrangement, it will be understood by those skilled in the art that modifications or equivalent substitutions may be made thereto without departing from the spirit and scope of the technical solution of the present invention.
Claims (10)
1. The ganoderma lucidum polysaccharide composition for regulating female intestinal flora is characterized by comprising the following raw materials in parts by weight:
ganoderma polysaccharide 25-50
Fructus Hippophae procyanidine 5-10
5-10 of mulberry leaf extract
Herba Houttuyniae extract 3-6
5-10 parts of raspberry fruit powder
5-10 of red date extract
3-6 parts of chia seed powder;
weighing the raw materials according to the weight part ratio, and uniformly mixing to obtain the ganoderma lucidum polysaccharide composition.
2. The ganoderan composition for regulating female intestinal flora according to claim 1, wherein the ganoderan composition comprises the following raw materials in parts by weight:
ganoderma lucidum polysaccharide 30-40
Fructus Hippophae procyanidine 6-8
5-7 parts of mulberry leaf extract
Herba Houttuyniae extract 3-4
8-10 parts of raspberry fruit powder
6-9 parts of red date extract
4-5 parts of chia seed powder.
3. The ganoderan composition for regulating female intestinal flora according to claim 1 or 2, characterized in that the ganoderan is extracted and purified by the method comprising: drying Ganoderma lucidum fruiting body at 40-70 deg.C, pulverizing, sieving with 50 mesh sieve, soaking in 8mL volume times of 95% ethanol for 24h according to 1g dry powder, removing lipid and small molecular substances, drying the filter residue under the above conditions, mixing uniformly according to the proportion of 10-30mL water of 1g powder, performing 500w ultrasonic treatment for 30min, performing water bath treatment at 40-90 deg.C for 60-120min, centrifuging to obtain supernatant, precipitating with 5 volume times of 95% ethanol for 4-8h, centrifuging to collect precipitate, dissolving in deionized water again according to 5mg/mL concentration, dialyzing with 5000Da bag for 72h, passing through Sephadex column to obtain single component, vacuum concentrating the obtained eluate, and freeze drying to obtain purified Ganoderma lucidum polysaccharide.
4. The ganoderan composition for modulating female gut flora as claimed in claim 1 or 2, wherein the seabuckthorn procyanidins are prepared by: taking the seabuckthorn seed meal after extracting the seabuckthorn seed oil as a raw material, carrying out flash extraction by using an ethanol solution at a low temperature, and filtering to obtain an extracting solution; recovering ethanol from the extract at low temperature and under negative pressure by using a heat pump device, and concentrating the extract to obtain extract; and (3) the extract is subjected to a belt drying device at a low temperature and under a negative pressure condition to obtain powdery seabuckthorn seed procyanidine.
5. The ganoderan composition for modulating female gut flora according to claim 1 or 2, wherein the mulberry leaf extract is prepared by the method comprising: cutting fresh mulberry leaves into strips, soaking the strips in acetic acid solution, filtering, collecting filtrate, centrifuging the obtained filtrate, collecting supernatant, concentrating, and drying to obtain solid to obtain the mulberry leaf extract.
6. The ganoderan composition for modulating the female gut flora of claim 1 or 2, wherein the houttuynia cordata extract is prepared by the method comprising: cutting fresh houttuynia cordata to obtain houttuynia cordata powder, decocting the powder with water under the condition of microwave and ultrasonic wave combined auxiliary dissolution, filtering to obtain houttuynia cordata extract, concentrating the extract under reduced pressure under the vacuum condition to obtain thick paste, and preparing the thick paste into the houttuynia cordata extract by a spray drying machine.
7. The ganoderan composition for regulating female intestinal flora according to claim 1 or 2, characterized in that the raspberry fruit powder is prepared by: cleaning fresh raspberry fruits without pedicels, crushing to prepare raspberry pulp, adding enzyme preparation, and performing enzymolysis at room temperature for 1-4h; passing the enzyme-hydrolyzed raspberry pulp through a homogenizer for 2-3 times to obtain homogenized raspberry pulp; and (3) passing the raspberry slurry through a high-pressure homogenizer to obtain the raspberry slurry after high-pressure homogenization, and carrying out low-temperature spray drying on the raspberry slurry to obtain the raspberry fruit powder.
8. The ganoderan composition for regulating female intestinal flora according to claim 1 or 2, characterized in that the preparation method of the red date extract comprises: taking dry red dates as raw materials, cleaning, soaking, crushing into red date pieces, adding an enzyme preparation, performing enzymolysis for 1-4 hours at 40-50 ℃, filtering to obtain supernatant, and performing vacuum concentration on the supernatant to obtain semi-solid red date paste; and (3) carrying out vacuum drying, crushing and sieving treatment on the semi-solid red date paste to obtain the red date extract.
9. Use of the ganoderan composition according to any one of claims 1 to 8 for the preparation of a food, a pharmaceutical or a nutraceutical for the prevention and/or regulation of disorders of the female intestinal flora.
10. The use of the ganoderan composition according to claim 9, wherein the food, drug or health product comprises other acceptable adjuvants in food, drug or health product, and is one of powder, tablet, granule, capsule or pill.
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