CN109771456A - Purposes of the trametes robinioplila extract in the preparation of preparation treatment and/or prevention intestinal bacilli illness disease and related disease - Google Patents
Purposes of the trametes robinioplila extract in the preparation of preparation treatment and/or prevention intestinal bacilli illness disease and related disease Download PDFInfo
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Abstract
The present invention relates to a kind of fungi extracts in the purposes of purposes more particularly to trametes robinioplila extract in the preparation of preparation treatment and/or prevention intestinal bacilli illness disease and related disease in the preparation of preparation treatment and/or prevention intestinal bacilli illness disease and related disease.Of the invention preparation method is simple, operating cost is low, and the Extraction solvent in preparation method of the invention can be with recycling and reusing, and no organic reagent remains risk, while ensure that the natural quality of trametes robinioplila extract and environmentally friendly.Raw material extraction is more complete, takes full advantage of raw material, and raw material availability is high.Trametes robinioplila extract can not only be clearly helpful for dominant microflora structure in enteron aisle and restore, facilitate flora diversification, but also significantly treatment and/or preventive effect are shown to disease relevant to intestinal microflora, it can be used in the various fields such as drug, health care product, there is great economy and society value.
Description
Technical field
The present invention relates to a kind of fungi extracts in preparation treatment and/or prevention intestinal bacilli illness disease and related disease
Preparation in purposes more particularly to trametes robinioplila extract in preparation treatment and/or prevention intestinal bacilli illness disease and related disease
Preparation in purposes.
Background technique
In recent years, it is continued to bring out about the research of intestinal flora, effect of the intestinal flora in human health is also increasingly
Obtain people's concern.Intestinal flora be human body intestinal canal all microbial totals and, many kinds of (including bacterium, fungi and disease
Poison), number is even more surprising.It is estimated that enteric bacteria total weight is up to 1~1.5kg in normal adult's body.They can
Weight, digestion and absorbability are influenced, the risk of infection and autoimmune disease etc. is resisted；In addition to this, also with it is some
The generation of endocrine metabolism disease, kidney trouble, the nervous system disease, cardiovascular disease, respiratory disease and cancer
With close association.
Intestinal flora combines according to a certain percentage, conditions each other, depends on each other for existence between each bacterium, and a kind of life is formed on quality and quantity
State balance, once body internal and external environment changes, for example including prolonged application broad-spectrum antibiotic, sensitive intestines bacterium be suppressed and not
Repressed bacterium seizes the opportunity breeding, and so as to cause flora imbalance, normal physiological combination is destroyed, to generate pathologic group
It closes, causes clinical symptoms, referred to as intestinal bacilli illness disease.Intestinal bacilli illness disease is not only harmful in itself, but also intestinal flora
Deficiency disorder can also bring some relevant diseases, including for example because hypoimmunity caused by intestinal bacilli illness disease, hyperlipidemia,
High cholesterol, obesity, road tissue damage, septicemia, inflammation such as intestinal inflammatory, colitis, neuroinflamation, tumour inflammation, mistake
It is quick etc..Therefore, the harm of intestinal bacilli illness disease and its related disease is extensive.
Currently, what is researched and developed both at home and abroad has there are mainly two types of the substances for adjusting intestinal flora function: first is that beneficial to living
Bacteria preparation, mainly based on Bifidobacterium and various lactic acid bacterias, the promotor second is that beneficial bacterium rises in value is made by this substance
The intrinsic bacterium proliferation of the physiological of body itself, forms with the enteron aisle ecological environment of beneficial bacterium dominance.
In ancient times, just using lactic acid bacteria come after fermented food, especially microscope invention, some bacteriums start gradually
It is utilized by people, there is the blank of probiotics (probiotics) from that point on.People for probiotics definition there is
Different views, but be substantially all and contain some basic characteristics of probiotics, i.e. probiotics and its product is able to maintain after
The health of host body inhibits the generation of defective mode.
Probiotic composition refers to the product by bacterium for producer's fermentation, such as Yoghourt, cheese etc..Probiotics further includes simple
The thallus of thallus, killed bacterial living and some enzymes of bacterial secretory and metabolite etc..Probiotics is usually some belongs to
Or the compound of bacterium composition is not belonged to, it also include the probiotics of single bacterium.Probiotics is mainly double on belonging to categorization levels
Discrimination Bacillus and Lactobacillus bacteria, Bifidobacterium mainly include bifidobacterium longum, bifidobacterium breve, bifidobacterium adolescentis, baby
Youngster Bifidobacterium；Lactobacillus mainly includes Lactobacillus casei and lactobacillus acidophilus etc. in enteron aisle.
Since discovery helicobacter pylori can cause gastric ulcer, people come to realise intestinal flora also to be had with disease
It closes.Scientist and tradespeople become more and more interested as the control strategy of drug therapy target to intestinal flora.Enterobacteriaceae
Group has plasticity, and compared with the change of host's Disease-causing gene, adjustment intestinal microflora seems more simple and effective, such as excrement
Transplanting；Regulate and control the relevant disease of intestinal flora by reasonable diet；Instruct reasonable employment antibiotic to maintain enteric microorganism
Stable state；Even fixed point administration, discovery drug novel targets etc..
Another substance prebiotics (Prebiotics) are had found along with probiotics, it is a kind of dietary supplements,
By the growth for stimulating the bacterium in a kind of or a small number of kind of bacterium colonies of selectivity and activity to host generate wholesome effect from
And improve the food composition that can not be digested of host health.Successful prebiotics should be the major part when passing through upper digestive tract
It is not digested and can be fermented by intestinal flora.Most importantly it only stimulates the growth of profitable strain, rather than has latent
In pathogenic or corrupt active harmful bacteria.Therefore, prebiotic-supplemented is also to improve Bacterial community, increase probiotics abundance
One of effective ways.
The research and development of healthy food need to comprehensively consider the nutritional need of human body and intestinal flora.Research finds adjustment processing work
Skill increases food particle volume and matrix strength, can reduce human body to the bioavailability of nutriment, increasing intestinal flora can
The nutriment utilized reaches the state of human body and flora two-win；Precursor substance in food passes through the conversion of flora, can be formed
The beneficial metabolics products such as short chain fatty acids, equol, urolithin, isothiocyanates, AHR ligand are the new incisions of food research and development
Point, but flora can also generate unwanted metabolic products；By contacting for enteron aisle and other organs, food may influence liver, painstaking effort
Pipe, nerve, behavior and state of mind etc..Therefore, food processing technology is adjusted, reaches human body and flora to the utilization of nutriment
It is modern food processing technology factor in need of consideration to two-win state.
FAO (Food and Agriculture Organization of the United Nation) (FAO) it is proposed that, the best dietary structure of the 21st century mankind be " meat or fish, one element, one
Mushroom ", " mushroom " therein are actually exactly the edible mushroom of multiple types.Edible mushroom is the food with extensive healthcare function and pharmacological activity
Savor nutritional supplement.A large amount of pharmacology and clinical research confirmation, the edible mushrooms such as ganoderma lucidum, Hericium erinaceus, grifola frondosus, tremella have both at home and abroad
The physiological functions such as immune, anticancer are adjusted, are the novel base-material and excellent food nutrition health-care additive of healthy food.
China is edible mushroom species diversity height country abundant, and sum statistics is up to more than 1000 kinds.Fungi is as food
With, medicinal or health care product, there is long history in China, great advantage and what is common is that having no toxic side effect.Modern medicine
Studies have shown that they be not only traditional QI invigorating, salubrity, get rid of illness, promoting menstruation, the functions such as lengthen one's life, also tool enhances human immunity, resists
Tumour, anti-aging and other effects, the healthy food based on food, medicinal fungus have occupied a tiny space in the international market.With
The raising of consumer health's quality requires more very the fining of health-oriented products, but the existing most function of edible and medical fungi health-oriented products
Energy range and the market propaganda are chaotic, function factor and quality control index etc. be indefinite, function not enough significantly cannot medicinalization, therefore,
It is badly in need of function factor (active site) definitely, with the quality control system for capableing of medicinalization effect and precision.
Summary of the invention
Against the above deficiency, the present invention provide a kind of trametes robinioplila extract treat and/or prevent intestinal bacilli illness disease and
New application in related disease, function is significant clear, can medicinalization, have practicability.
The present invention reaches above-mentioned purpose by following scheme:
In the first aspect of the present invention, provide a kind of trametes robinioplila extract treat and/or prevent intestinal bacilli illness disease and
Purposes in related disease.
In the second aspect of the present invention, a kind of trametes robinioplila extract is provided in preparation treatment and/or prevention intestinal bacilli illness
Purposes in the preparation of disease and related disease.
In the third aspect of the present invention, a kind of system treated and/or prevent intestinal bacilli illness disease and related disease is provided
Agent, the preparation include trametes robinioplila extract.
In the fourth aspect of the present invention, a kind of medicine prevented and/or treat intestinal bacilli illness disease and related disease is provided
Object includes trametes robinioplila extract and pharmaceutically acceptable carrier.
In the fifth aspect of the invention, a kind of preparation method of trametes robinioplila extract is provided, comprising: crush trametes robinioplila fructification
Afterwards, it is successively mentioned with alcohol extracting, water, the extraction of sodium hydroxide (NaOH aqueous solution) buck, respectively obtains extraction solution, then respectively through dense
Contracting, freeze-drying, obtain trametes robinioplila alcohol extracting thing, trametes robinioplila water extract, trametes robinioplila alkali extract.
Compared with prior art, the invention has the following beneficial effects:
(1) of the invention preparation method is simple, operating cost is low, and the Extraction solvent in preparation method of the invention is only
Use ethyl alcohol, water, NaOH buck, ethyl alcohol that can remain risk with recycling and reusing, no organic reagent, ensure that trametes robinioplila extract
Natural quality while and it is environmentally friendly, belong to green manufacture technology.
(2) present invention successively uses ethyl alcohol, second alcohol and water, and NaOH buck extracts trametes robinioplila fructification, and raw material extraction is more complete,
Raw material is taken full advantage of, raw material availability is high.
(3) trametes robinioplila extract of the invention can not only be clearly helpful for dominant microflora structure recovery in enteron aisle, facilitate bacterium
Group's diversification, but also significantly treatment and/or preventive effect are shown to disease relevant to intestinal microflora, such as can
The deterioration of inflammation is effectively relieved, improves immune-related every physiopathological index, can be used for the various necks such as drug, health care product
In domain, there is great economy and society value.
Detailed description of the invention
Fig. 1 is small intestine HE coloring pathological section.
Specific embodiment
In the first aspect of the present invention, provide a kind of trametes robinioplila extract treat and/or prevent intestinal bacilli illness disease and
Purposes in related disease.
Wherein, the intestinal bacilli illness disease includes the intestinal bacilli illness disease as caused by improper diet.
Wherein, the related disease of the intestinal bacilli illness disease includes but is not limited to because being immunized caused by intestinal bacilli illness
Power is low, the damage of hyperlipidemia, high cholesterol, obesity, intestinal tissue, septicemia, inflammation, allergy.
Further, above-mentioned inflammation includes but is not limited to intestinal inflammatory, colitis, neuroinflamation, tumour inflammation.
In the second aspect of the present invention, a kind of trametes robinioplila extract is provided in preparation treatment and/or prevention intestinal bacilli illness
Purposes in the preparation of disease and related disease.
Wherein, the intestinal bacilli illness disease includes the intestinal bacilli illness disease as caused by improper diet.
Wherein, the related disease of the intestinal bacilli illness disease includes but is not limited to because being immunized caused by intestinal bacilli illness
Power is low, the damage of hyperlipidemia, high cholesterol, obesity, intestinal tissue, septicemia, inflammation, allergy.
Further, above-mentioned inflammation includes but is not limited to intestinal inflammatory, colitis, neuroinflamation, tumour inflammation.
For example, in one embodiment, a kind of trametes robinioplila extract is provided and is preparing the purposes in immunopotentiator.
In the third aspect of the present invention, a kind of system treated and/or prevent intestinal bacilli illness disease and related disease is provided
Agent, the preparation include trametes robinioplila extract.
Preferably, the preparation includes drug, health care product or food.
Wherein, the intestinal bacilli illness disease includes the intestinal bacilli illness disease as caused by improper diet.
Wherein, the related disease of the intestinal bacilli illness disease includes but is not limited to because being immunized caused by intestinal bacilli illness
Power is low, the damage of hyperlipidemia, high cholesterol, obesity, intestinal tissue, septicemia, inflammation, allergy.
Further, above-mentioned inflammation includes but is not limited to intestinal inflammatory, colitis, neuroinflamation, tumour inflammation.
In the fourth aspect of the present invention, a kind of medicine prevented and/or treat intestinal bacilli illness disease and related disease is provided
Object includes trametes robinioplila extract and pharmaceutically acceptable carrier.
Wherein, the intestinal bacilli illness disease includes the intestinal bacilli illness disease as caused by improper diet.
Wherein, the related disease of the intestinal bacilli illness disease includes but is not limited to because being immunized caused by intestinal bacilli illness
Power is low, the damage of hyperlipidemia, high cholesterol, obesity, intestinal tissue, septicemia, inflammation, allergy.
Further, above-mentioned inflammation includes but is not limited to intestinal inflammatory, colitis, neuroinflamation, tumour inflammation.
Heretofore described " pharmaceutically acceptable carrier " is being cured it is also understood that for " diluent " or " auxiliary material " etc.
It is known that and being described in such as Remington medical science (Remington ' s Pharmaceutical in medicine field
Sciences), Mack Publishing Company (Mack Publishing Co.) (AR lattice nano (A.R.Gennaro) compile,
1985).These substances are nontoxic for recipient under used dosage and concentration, including such as phosphate, citric acid
The buffers such as salt, acetate and other acylates；The antioxidants such as ascorbic acid；The low molecular weights such as poly arginine
(being less than about 10 residues) peptide；The protein such as seralbumin, gelatin or immunoglobulin；Such as polyvinylpyrrolidone
Equal hydrophilic polymers；The amino acid such as glycine, glutamic acid, aspartic acid or arginine；Monosaccharide, disaccharides and including cellulose
Or derivatives thereof, other carbohydrate of glucose, mannose or dextrin；The chelating agents such as EDTA；Such as mannitol or
The sugar alcohols such as sorbierite；The counter ion counterionsl gegenions such as sodium；And/or such as tween (Tween), Pluronic (Pluronics) or poly- second
The nonionic surface active agent such as glycol；Further including includes sterile water, salt water, glucose, oil (such as corn oil, peanut oil and class
Like object), acid.Further can also include preservative, wetting agent, emulsifier, and the like etc..
The dosage of trametes robinioplila extract or drug of the invention can with specific medication and treated object or
Type and change.The dosage is enough to generate expected beneficial effect, and can be formulated for single dose application or multiple dose
Amount application.
Heretofore described drug can be applied by any approach well known by persons skilled in the art, including oral,
The approach such as intramuscular, intravenous, intradermal, intralesional.Application can be part, surface or whole body.In any given feelings
Most suitable approach depends on many factors, such as property, the process of disease, the seriousness of disease of disease etc. under condition.As
It is preferred that trametes robinioplila extract of the invention or being administered orally comprising the drug including trametes robinioplila extract.
The trametes robinioplila extract of the invention can also be applied together with the second reactive compound/second therapeutic agent, simultaneously,
It applies in succession or parallel.When applying respectively, the trametes robinioplila extract of the invention or the drug comprising trametes robinioplila extract and
Two reactive compounds or second therapeutic agent can be applied with different medicine frequencies or interval.
In the purposes of aforementioned four aspect of the invention or preparation, the trametes robinioplila extract include trametes robinioplila water extract,
Trametes robinioplila alcohol extracting thing, trametes robinioplila alkali extract.
In one embodiment, the trametes robinioplila alcohol extracting thing is trametes robinioplila ethanol extract.
In one embodiment, the trametes robinioplila alkali extract is trametes robinioplila sodium hydroxide extract.
It is furthermore preferred that in the purposes of four aspects of aforementioned present invention or preparation, under the trametes robinioplila extract is preferably
State the trametes robinioplila extract that preparation method is prepared.
In the fifth aspect of the invention, a kind of preparation method of trametes robinioplila extract is provided, comprising: crush trametes robinioplila fructification
Afterwards, it is successively mentioned with alcohol extracting, water, the extraction of NaOH buck, respectively obtains extraction solution, then concentrated, freeze-drying respectively, obtain Chinese scholartree
Ear alcohol extracting thing, trametes robinioplila water extract, trametes robinioplila alkali extract.
Preferably, a kind of preparation method of trametes robinioplila extract, comprising: after crushing trametes robinioplila fructification, successively with 95% second
Alcohol extracting, water are extracted, 1%-2%NaOH buck extracts, and respectively obtain extraction solution, then concentrated, freeze-drying respectively, are obtained
Trametes robinioplila alcohol extracting thing, trametes robinioplila water extract, trametes robinioplila alkali extract.
Preferably, a kind of preparation method of trametes robinioplila extract, comprising: after crushing trametes robinioplila fructification, using alcohol reflux
It extracts, obtains the first filter residue and alcohol extracting solution, the first filter residue is used into water refluxing extraction solution, the second filter residue is obtained and water mentions
Solution is taken, then the second filter residue is used into 1%-2%NaOH buck refluxing extraction, third filter residue is obtained and alkali carries takes solution, by alcohol
It extracts solution, water extraction solution and obtains trametes robinioplila alcohol extracting thing, trametes robinioplila water extract by concentration, freeze-drying respectively；By alkali carries
Solution is taken to obtain trametes robinioplila alkali extract by concentration, alcohol precipitation, freeze-drying.
Preferably, it includes: that the ethyl alcohol that solid-liquid ratio is 1:8-1:15 is added that the alcohol reflux, which extracts, and refluxing extraction 2-4 is small
When, it filters, obtains filtrate, is concentrated under reduced pressure to give thick paste, -80 DEG C of freeze overnights of thick paste warp are freeze-dried to get trametes robinioplila alcohol extracting
Object.
Preferably, the water refluxing extraction includes: the filter residue after taking alcohol extracting, uses solid-liquid ratio for the water of 1:8-1:15, is returned
Stream extracts 1-3 hours, filters, obtains filtrate, is concentrated under reduced pressure to give thick paste, -80 DEG C of freeze overnights of thick paste warp, freeze-drying, i.e.,
Obtain trametes robinioplila water extract.
Preferably, the 1%-2%NaOH buck extract include: learn from else's experience alcohol and water mention after filter residue, use solid-liquid ratio for
The buck of 1:8-1:15, stirring is ultrasonic extraction 1-2 hours lower, filters, and obtains filtrate, and alcohol reagent precipitating is added, precipitates 12-24
Hour, precipitating is collected, through -80 DEG C of freeze overnights, is freeze-dried to get trametes robinioplila alkali carries object.
The 1%-2%NaOH or the 1%-2%NaOH buck refer to that weight percentage is the NaOH water of 1%-2%
Solution.
Preferably, the 1%-2%NaOH is 2%NaOH.
It is further preferred that the alcohol reagent of the precipitating includes ethyl alcohol, methanol.
It is further preferred that described be precipitated as that 4-6 times of the liquid ethyl alcohol measured is added, precipitate 12-24 hours.
In the sixth aspect of the present invention, a kind of trametes robinioplila extract that above-mentioned preparation method is prepared is provided.
Trametes robinioplila (ew'sEaronPagodaTCMLIBee), be On Polyporaceae Chinese scholartree bolt bacterium fructification, alias: Chinese scholartree Xiao,
Chinese scholartree bacterium, Chinese scholartree chicken, Chinese scholartree goose, Chinese scholartree moth, red chicken.Chinese scholartree bolt bacterium Trametes robiniophila Murr.Fructification stockless, cap semicircle
Shape is often in imbricate, and suberin, sepia, dipped beam is sliding, there is a small number of ring grains, and bacterial context yellow-white is savory after doing, wall thickness and light
Whole, aperture yellow-white, polygonal, 5~6 between every 1mm, spore is colourless, smooth, and spore print white often has cystidium.It is grown on
On the trunks such as Chinese scholartree and acacia, wingceltis.It is distributed in the ground such as Hebei, Shandong, Shaanxi, Sichuan, Chongqing.Wild resource is rare, Jiangsu etc.
Ground uses solid fermentation method culture Chinese scholartree bolt bacterium with hyoscine.Has the effects that antitumor, enhancing immune function and antiviral.
Heretofore described " imbalance " can be understood as " unbalance ", " disorder " or " disease " etc..
As used herein, term "comprising" or " comprising " refer to that method includes cited element, but are not excluded for other
Element.
Trametes robinioplila of the present invention is the fructification of On Polyporaceae Chinese scholartree bolt bacterium, and principal component is trametes robinioplila proteoglycan,
Hydrolysate (fucose) containing L-fucose, L-arabinose (arabinose), D- xylose (xylose), D-MANNOSE
(mannose), D- galactolipin (galactose), 6 kinds of monosaccharide such as D-Glucose (glucose) and aspartic acid (aspartic
Acid), threonine (threonine), serine (serine), glutamic acid (glutamicacid), proline (proline),
Glycine (glycine), alanine (alanine), cystine (cystine), valine (valine), methionine
(methionine), isoleucine (isoleucine), leucine (leucine), tyrosine (tyrosine), phenylalanine
(phenylalanine), lysine (lysine), histidine (histidine), tryptophan (Typtophane), arginine
(arginine) 18 kinds of amino acid compositions such as.This edible mushroom value is high.
Below in conjunction with specific embodiment, invention is further explained.
Embodiment 1: the preparation of trametes robinioplila extract, steps are as follows:
(1) trametes robinioplila fructification is crushed, uses solid-liquid ratio for the ethyl alcohol of 1:8-1:15, refluxing extraction 2-4 hours, filtered,
The thick paste for obtaining filtrate, being concentrated under reduced pressure to give debita spissitudo, -80 DEG C of freeze overnights of thick paste warp, enter freeze drier it is dry to get
Trametes robinioplila alcohol extracting thing.
(2) the above-mentioned filter residue after alcohol extracting is taken, uses solid-liquid ratio for the water of 1:8-1:15, refluxing extraction 1-3 hours, is taken out
Filter, the thick paste for obtaining filtrate, being concentrated under reduced pressure to give debita spissitudo, -80 DEG C of freeze overnights of thick paste warp enter freeze drier drying,
Up to trametes robinioplila water extract.
(3) the above-mentioned filter residue after alcohol and water mentions is taken, uses solid-liquid ratio for the 2%NaOH aqueous solution of 1:8-1:15, is stirred
Lower ultrasonic extraction 1-2 hours filters, and obtains filtrate, and debita spissitudo ethyl alcohol is added, and precipitates 12-24 hours；Precipitating is collected, through -80
It is dry to get trametes robinioplila buck extract to enter freeze drier for DEG C freeze overnight.
Determination of polysaccharide uses People's Republic of China's agricultural industry criteria NY/T 1676-2008 " thick more in edible mushroom
The measurement of sugared content " method；Protein content determination uses Kjeldahl's method.Thick many candies and protein content in trametes robinioplila extract
As shown in table 1.
Thick many candies and protein content in 1 trametes robinioplila extract of table
Embodiment 2: trametes robinioplila extract causes the improvement result of mouse intestinal flora imbalance index of correlation to high fat diet
1, reagent:
Total cholesterol (T-CHO) testing cassete: Bioengineering Research Institute is built up in Nanjing.
Triglycerides (TG) testing cassete: Bioengineering Research Institute is built up in Nanjing.
Low density lipoprotein cholesterol (LDL-C) testing cassete: Bioengineering Research Institute is built up in Nanjing.
High-density lipoprotein cholesterol (HDL-C) testing cassete: Bioengineering Research Institute is built up in Nanjing.
Female C57 mouse 30, is purchased from Guangdong Province's medical experiment animal by SPF grades, weight 12-14g, age in days 28-30d
Center, credit number are SCXK (Guangdong) 2013-0002.
Rats and mice maintains feed: Guangdong Medical Lab Animal Center, credit number SCXK (Guangdong) 2013-0002.
High cholesterol diet formula high in fat is as shown in table 2.
The main component and energy of the high cholesterol diet high in fat of table 2 are constituted
Trametes robinioplila extract is prepared by experimental example 1 of the present invention.
2, method
The preparation of high fat diet obesity-induced mice model: animal subject is raised in the zoopery of Institute of Micro-biology, Guangdong Province
The heart, raising temperature and humidity: 23 ± 1 DEG C, 55 ± 10%, using 12h intermittent illumination round the clock；Receptacle condition remains steady
It is fixed, with the reliability of guarantee test result.Mouse ad lib drinking-water.It is primary that period is checked daily animal, does not find unhealthy
Animal, take whole healthy mices to be tested.50 C57 mouse are randomly divided into normal group, model after adaptive feeding 7d
Group, trametes robinioplila alcohol extracting group, trametes robinioplila water propose group and trametes robinioplila alkali carries group, and every group 10.Normal group is fed with normal diet, model group and is given
Medicine group is fed with high cholesterol diet high in fat, continuous to feed 2 months.
After modeling success, start to be administered, administration group gives trametes robinioplila water extract, trametes robinioplila alcohol extracting thing and trametes robinioplila alkali carries respectively
Take object stomach-filling, drug concentration 6mg/ml (180g extract is dissolved in 30ml water), 3mg/ pcs/day of dosage, dosage 0.5ml/ only/
It, successive administration 35 days, while normal group mouse continues to be fed with normal diet, administration group and model group mouse use common feeding instead
Material is fed.
3, result is observed
The appearance posture (hair, color, shape) and dynamic of daily observation animal simultaneously make respective record, measure the weight of animals every 7d
1 time.
Blood: after being discontinued for 24 hours, animal materials testing index.Acquire blood sample, pluck eyeball and take blood, blood plasma through 3000RPM,
Serum is collected in 10min centrifugation twice, measures total cholesterol (TG), triglycerides (T-CHO), low-density lipoprotein in serum
(LDL-C) and high-density lipoprotein (HDL-C) is horizontal.Small intestine: it plucks after eyeball takes blood, puts to death and dissect mouse, take mouse
Small intestine is fixed in 4% paraformaldehyde.Small intestine is made into paraffin section, is dyed using HE method, carries out pathological section
Observation.
4, data processing
All data are with mean ± standard deviationIt indicates.The comparison of mean uses one-way analysis of variance between multiple groups
(One-Way ANOVA), mean compares two-by-two between group, and LSD method is used when variance is neat；Dunnett ' s T3 is used when heterogeneity of variance
Method.It is completed by SPSS software, α=0.05.
The changes of weight of each group is as shown in table 3.
The variation of 3 mouse weight of table
#P < 0.05,##P < 0.01,###Normal group of P < 0.001vs；*P < 0.05,**P < 0.01,***P < 0.001vs model group
It has been observed that the fur of the animal of administration group is more smoother than the fur of model group.
From table 3 it can be seen that the fur of the animal by treatment is more smoother than the fur of model group.Normal group and model group
Between average weight significant difference (P < 0.05), model group mouse start when average weight about 65.45g, terminate in experiment
When average weight be 72.76g, illustrate high fat diet obesity-induced mice model modeling success；Compared with model group, 3 administrations
Group average mice body weight is remarkably decreased (P < 0.001) (table 3).This shows: can to the suitable trametes robinioplila extract of obesity mice stomach-filling
To significantly slow weight gain.Especially trametes robinioplila alcohol extract slows down weight significant effect.
The four items of blood lipid tests testing result of each group is as shown in table 4.
4 mice serum biochemical indicator (mmol/L) of table
#P < 0.05,##P < 0.01,###Normal group of P < 0.001vs；*P < 0.05,**P < 0.01,***P < 0.001vs model group
From table 4, it can be seen that in model group total cholesterol, triglycerides and ldl-cholesterol ratio normally organize it is aobvious
It writes and increases (P < 0.01 or P < 0.001), high-density lipoprotein cholesterol significantly reduces (P < 0.001) than normal group；With model group
It compares, triglycerides, total cholesterol, low-density lipoprotein cholesterol level are remarkably decreased (P < 0.05 or P < 0.01 in administration group
Or P < 0.001), High-density Lipoprotein-cholesterol significantly increases (P < 0.05).This shows: suitable to obesity mice stomach-filling
Total cholesterol, triglycerides and low-density lipoprotein white level in serum can be significantly reduced in trametes robinioplila extract.Especially trametes robinioplila
The effect of water extract is particularly significant.
Small intestine HE coloring pathological section result as shown in Figure 1, it can be seen from figure 1 that normally group small intestine's villus it is long and
Completely, model group intestinal villus falls off seriously, illustrates that high fat diet can cause to damage to the enteron aisle of mouse, illustrates that model is success
's.Compared with model group, after drug-treated, administration group can alleviate damage, promote the reparation of enteron aisle.
Embodiment 3: improvement result of the trametes robinioplila extract to high fat diet obesity-induced mice intestinal bacilli illness
1, experimental method
Stool in mice is stored in -80 DEG C of ice after liquid nitrogen is rapidly frozen at the end of zoopery in acquisition embodiment 2
In case.DNA is extracted from fecal specimens, and DNA is detected by 0.8% agarose gel electrophoresis and extracts quality, while using purple
Outer spectrophotometer quantifies DNA.Using forward primer 5 '-ACTCCTACGGGAGGCAGCA-3 ' and reverse primer 5'-
GGACTACHVGGGTWTCTAAT-3' expands the area the V3-V4 gene of mouse 16S rRNA.Pcr amplification product passes through 2%
Agarose gel electrophoresis is detected, and carries out gel extraction to target fragment, and recycling is recycled using the gel of AXYGEN company
Kit.Referring to the preliminary quantitative result of electrophoresis, PCR amplification recovery product is subjected to fluorescent quantitation, fluorescent reagent Quant-iT
PicoGreen dsDNA Assay Kit, quantitative instrument are Microplate reader (BioTek, FLx800).According to fluorescence
Quantitative result is mixed each sample by corresponding proportion according to the sequencing amount demand of each sample.Using Illumina company
TruSeq Nano DNA LT Library Prep Kit prepare sequencing library.2 × 300bp is carried out using MiSeq sequenator
Both-end sequencing, corresponding reagent be MiSeq Reagent Kit V3 (600cycles).
1, experimental result
The results are shown in Table 5 for the intestinal flora diversity of each group.
5 mouse intestinal flora diversity indices of table
#P < 0.05,##P < 0.01,###Normal group of P < 0.001vs；*P < 0.05,**P < 0.01,***P < 0.001vs model group
As can be seen from Table 5, compared with normal group, the Alpha diversity indices Chao1 of model group mouse intestinal flora,
Shannon and Simpson is remarkably decreased (P < 0.05), illustrates that high fat diet changes mouse intestinal flora structure, makes its multiplicity
Property reduce；Compared with model group, administration group mouse intestinal flora diversity indices ACE, Chao1, Shannon and Simpson are aobvious
It writes and increases (P < 0.05), after illustrating that the obesity mice intestinal flora of high fat diet induction gives trametes robinioplila extract for treating, enterobacteriaceae
Group's diversity dramatically increases.The OTU enrichment analysis of each group is as shown in table 6 and table 7.
Relative abundance of 6 mouse intestinal flora of table in door level
Group；*P < 0.05,**P < 0.01,***P < 0.001vs model group
7 mouse intestinal flora of table is belonging to the relative abundance in level
#P < 0.05,##P < 0.01,###Normal group of P < 0.001vs；*P < 0.05,**P < 0.01,***P < 0.001vs model group
As can be seen from Table 6, in door level, compared with normal group, model group mouse intestinal flora Bacteroidetes
Significant changes have occurred with Firmicutes relative abundance, Bacteroidetes relative abundance significantly reduces (P < 0.05),
Firmicutes relative abundance dramatically increases (P < 0.05)；Compared with model group, administration group Bacteroidetes relative abundance is aobvious
It writes and increases (P < 0.05 or P < 0.01), Firmicutes relative abundance significantly reduces (P < 0.05 or P < 0.01).
As can be seen from Table 7, on belonging to level, compared with normal group, model group mouse intestinal flora Bacteroides,
Lactobacillus and Odoribacter relative abundance significantly reduces (P < 0.05), and Ruminococcus relative abundance significantly increases
Add (P < 0.05)；Compared with model group, administration group Bacteroides, Lactobacillus and Odoribacter are dramatically increased
(P < 0.05), Ruminococcus relative abundance significantly reduce (P < 0.05).
This explanation, trametes robinioplila extract of the invention have the function of adjusting intestinal bacilli illness.
To sum up it can be seen that, trametes robinioplila extract can not only be clearly helpful for dominant microflora structure in enteron aisle and restore, and facilitate
Flora diversification, but also significantly treatment and/or preventive effect are shown to disease relevant to intestinal microflora, can have
Effect alleviates the deterioration of inflammation, improves immune-related every physiopathological index, can be used for the various fields such as drug, health care product
In, there is great economy and society value.
The above, preferable specific embodiment only of the invention, but scope of protection of the present invention is not limited thereto,
Anyone skilled in the art in the technical scope disclosed by the present invention, according to the technique and scheme of the present invention and its
Design is subject to equivalent substitution or change, should be covered by the scope of protection of the present invention.
Claims (10)
1. purposes of the trametes robinioplila extract in the preparation of preparation treatment and/or prevention intestinal bacilli illness disease and related disease.
2. purposes according to claim 1, which is characterized in that the related disease of the intestinal bacilli illness disease includes but not
Hypoimmunity caused by being limited to because of intestinal bacilli illness, hyperlipidemia, high cholesterol, obesity, intestinal tissue damage, septicemia,
Inflammation, allergy；Preferably, the inflammation includes but is not limited to intestinal inflammatory, colitis, neuroinflamation, tumour inflammation.
3. purposes according to claim 1, which is characterized in that trametes robinioplila extract is preparing the purposes in immunopotentiator.
4. the preparation of a kind for the treatment of and/or prevention intestinal bacilli illness disease and related disease, which is characterized in that the preparation includes
Trametes robinioplila extract.
5. preparation according to claim 4, which is characterized in that the related disease of the intestinal bacilli illness disease includes but not
Hypoimmunity caused by being limited to because of intestinal bacilli illness, hyperlipidemia, high cholesterol, obesity, intestinal tissue damage, septicemia,
Inflammation, allergy；Preferably, the inflammation includes but is not limited to intestinal inflammatory, colitis, neuroinflamation, tumour inflammation.
6. the drug of a kind of prevention and/or treatment intestinal bacilli illness disease and related disease, which is characterized in that extracted comprising trametes robinioplila
Object and pharmaceutically acceptable carrier.
7. drug according to claim 6, which is characterized in that the trametes robinioplila extract includes trametes robinioplila water extract, trametes robinioplila
Alcohol extracting thing, trametes robinioplila alkali extract.
8. a kind of preparation method of trametes robinioplila extract characterized by comprising by trametes robinioplila fructification crush after, successively with alcohol extracting,
Water mentions, NaOH buck extracts, and respectively obtains extraction solution, then concentrated, freeze-drying respectively, obtains trametes robinioplila alcohol extracting thing, Chinese scholartree
Ear water extract, trametes robinioplila alkali extract.
9. preparation method according to claim 8 characterized by comprising after crushing trametes robinioplila fructification, successively use
95% ethyl alcohol extracts, water extracts, 1%-2%NaOH buck extracts, and respectively obtains extraction solution, then concentrated, freezing is done respectively
It is dry, obtain trametes robinioplila alcohol extracting thing, trametes robinioplila water extract, trametes robinioplila alkali extract.
10. preparation method according to claim 8 characterized by comprising after crushing trametes robinioplila fructification, using second
Alcohol reflux extracts, and obtains the first filter residue and alcohol extracting solution, and the first filter residue is used water refluxing extraction solution, obtains the second filter residue
And water extraction solution, then the second filter residue is used into 1%-2%NaOH buck refluxing extraction, obtain third filter residue and alkali carries take it is molten
Alcohol extracting solution, water extraction solution are obtained trametes robinioplila alcohol extracting thing, trametes robinioplila water extract by concentration, freeze-drying respectively by liquid；
Solution is taken to obtain trametes robinioplila alkali extract by concentration, alcohol precipitation, freeze-drying alkali carries.
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