CN109876020A - Purposes of the cicada fungus buck extract in the preparation of preparation treatment and/or prevention intestinal bacilli illness disease and related disease - Google Patents
Purposes of the cicada fungus buck 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 cicada fungus buck 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 has only used buck, and no organic reagent remains risk, while ensure that the natural quality of cicada extract and environmentally friendly, belongs to green manufacture technology.Cicada fungus buck extract of the invention 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, such as the deterioration of inflammation can be effectively relieved, improve immune-related every physiopathological index, 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 cicada fungus buck extract in preparation treatment and/or prevention intestinal bacilli illness disease and correlation
Purposes in the preparation of disease.
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. are indefinite, function not enough significantly cannot medicinalization.
Therefore, it is badly in need of function factor (active site) definitely, with capableing of medicinalization effect and precision
Quality control system.
Summary of the invention
Against the above deficiency, the present invention provides a kind of cicada fungus buck extract and is treating and/or preventing intestinal bacilli illness
New application in disease and 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, a kind of cicada fungus buck extract is provided and is treating and/or preventing intestinal bacilli illness
Purposes in disease and related disease.
In the second aspect of the present invention, a kind of cicada fungus buck extract is provided in preparation treatment and/or prevention intestinal flora
Purposes in deficiency disorder and the preparation of 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 cicada fungus buck 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 cicada fungus buck extract and pharmaceutically acceptable carrier.
In the fifth aspect of the invention, a kind of preparation method of cicada fungus buck extract is provided, comprising:
After cicada fungus fructification is crushed, the aqueous alkali measured with 12-16 times ultrasonic extraction 1-2 hours, obtains extracting molten
Liquid is concentrated under reduced pressure, and adjusts pH to 4~5, and precipitating is precipitated in refrigeration, takes pellet frozen dry, obtains cicada fungus buck 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
Using buck, no organic reagent remains risk, and it is while ensure that the natural quality of cicada extract and environmentally friendly, belong to
In green manufacture technology.
(2) present invention extracts cicada fungus fructification using buck, and raw material extraction is more complete, takes full advantage of raw material.
(3) cicada fungus buck extract of the invention can not only be clearly helpful for dominant microflora structure recovery in enteron aisle, help
Significantly treatment and/or preventive effect, example are shown in flora diversification, but also to disease relevant to intestinal microflora
As can the deterioration of inflammation is effectively relieved, improves immune-related every physiopathological index, it is each to can be used for drug, health care product etc.
In kind field, there is great economy and society value.
Detailed description of the invention
Fig. 1 is the small intestine HE coloring pathological section figure (observation of 200 power microscopes) of embodiment 2.
Specific embodiment
In the first aspect of the present invention, a kind of cicada fungus buck is provided, object is taken to treat and/or preventing intestinal bacilli illness disease
And the 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 cicada fungus buck extract is provided in preparation treatment and/or prevention intestinal flora
Purposes in deficiency disorder and the preparation of 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 cicada fungus buck 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 cicada fungus buck 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 cicada fungus buck 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 cicada fungus buck extract or drug of the invention can be with specific medication and treated visitor
Body or type and change.The dosage is enough to generate expected beneficial effect, and can be formulated for single dose application or more
Relatively heavy 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 cicada fungus buck extract of the invention or being administered orally comprising the drug including cicada fungus buck extract.
The cicada fungus buck 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 cicada fungus buck extract of the invention or include cicada fungus buck extract
The drug of object can be applied from the second reactive compound or second therapeutic agent with different medicine frequencies or interval.
It is furthermore preferred that the cicada fungus buck extract is preferably following preparations in the purposes and preparation of aforementioned four aspect
The cicada fungus buck extract that method is prepared.
In the fifth aspect of the invention, a kind of preparation method of cicada fungus buck extract is provided, comprising:
After cicada fungus fructification is crushed, the aqueous alkali measured with 12-16 times ultrasonic extraction 1-2 hours, obtains extracting molten
Liquid is concentrated under reduced pressure, and adjusts pH to 4~5, and precipitating is precipitated in refrigeration, takes pellet frozen dry, obtains cicada fungus buck extract.
Preferably, the aqueous alkali be include NaOH and/or NaHCO that pH is 9-103Aqueous solution.
Preferably, the tune pH is to add acetic acid tune pH.
Preferably, the refrigeration is refrigeration 24 hours.
A kind of preparation method of cicada fungus buck extract, comprising:
After cicada fungus fructification is crushed, the pH measured with 12-16 times is 9~10 sodium hydroxide aqueous alkalis, ultrasonic extraction 1-2
Hour, it obtains extracting solution, be concentrated under reduced pressure, add acetic acid tune pH to 4~5, refrigerate 24 hours, precipitating is precipitated, take pellet frozen dry
It is dry, obtain extract.
In the sixth aspect of the present invention, a kind of cicada fungus buck extract that above-mentioned preparation method is prepared is provided.
Cicada fungus (Isaria cicadae Miquel) is commonly called as big cordyceps sinensis, belongs to entomogenous fungi;The long kidney shape of sclerotium shape, it is micro-
Bending, is about 2.5-3.5cm, diameter 1-1.4cm is similar to the larva of cicada.Polypide head has 1-2 pieces of stroma and is also referred to as falx
Beam, strip or curling, branch or not branch, long 3-7cm, diameter 3-4mm, ecosystem egg white color, milky after drying also have
For dark brown, top is slightly expanded, and there is powdery cicada fungus conidia powder on surface, is similar to flower.Coremium is also referred to as fructification, stroma, by posting
Main front end is born, white when fresh, high 1.5-6cm;Handle branch or not branch, diameter 0.1-0.2cm.Cicada fungus mainly appears on
Sichuan, the Jiangsu, Zhejiang, Fujian of China, but Anhui, Yunnan Province east are distributed allly.With improve immunity, resist it is tired
Labor protects kidney, improves the multiple actions such as sleep, antitumor, liver protection, anti-radiation and improving eyesight, is magical ancient Chinese medicine.
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.
Cicada fungus of the present invention is containing mannitol, polysaccharide, amino acid, cordycepin, adenosine etc., with natural cordyceps phase
Seemingly.Below in conjunction with specific embodiment, invention is further explained.
Below in conjunction with specific embodiment, invention is further explained.
Embodiment 1: the preparation method of cicada fungus buck extract, steps are as follows:
After cicada fungus fructification is crushed, with 12-16 times of 9~10 sodium hydroxide aqueous alkali of the pH measured, ultrasonic extraction 1-2
Hour, it obtains extracting solution, is concentrated under reduced pressure into proper volume plus acetic acid tune pH to 4~5, refrigerate 24 hours, precipitating is precipitated, takes
Pellet frozen is dried, and obtains cicada fungus buck extract.
Protein content determination uses Kjeldahl's method.Table 1 is protein content in cicada fungus buck extract.
Protein content in 1 cicada fungus buck extract of table
Protein (%) | |
Cicada fungus buck extract group | 55.55±2.31 |
Embodiment 2: cicada fungus buck 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 28-30 days
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
Cicada 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 12 hours intermittent illuminations round the clock;Receptacle condition remains
Stablize, with the reliability of guarantee test result.Mouse ad lib drinking-water.It is primary that period is checked daily animal, does not find not to be good for
The animal of health takes whole healthy mices to be tested.30 C57 mouse after adaptive feeding 7 days, be randomly divided into normal group,
Model group, cicada fungus buck extract group, every group 10.Normal group is fed with normal diet, model group and administration group height high in fat
Cholesterol feed is fed, continuous to feed 2 months.
After modeling success, start to be administered, administration group gives cicada fungus buck extract, and (180g is extracted drug concentration 6mg/ml
Object is dissolved in 30ml buck), 3mg/ pcs/day of dosage, 0.5ml/ pcs/day of dosage, successive administration 35 days, while normally organizing mouse
Continue to be fed with normal diet, administration group and model group mouse use normal diet nursing instead.
3, result is observed
The appearance posture (hair, color, shape) and dynamic of daily observation animal simultaneously made respective record, every 7 days measurement animal bodies
It weighs 1 time.
Blood: after being discontinued 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, and it is solid to measure total cholesterol (T-CHO), triglycerides (TG), low-density lipoprotein gallbladder in serum
Alcohol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) are horizontal.
Small intestine: it plucks after eyeball takes blood, puts to death and dissect mouse, take mouse small intestine tissue, be fixed on 4% paraformaldehyde
In.Small intestine is made into paraffin section, is dyed using HE method, carries out pathology section examination.
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.
It has been observed that the fur of the animal of administration group is more smoother than the fur of model group.
The changes of weight of each group is as shown in table 3.
The variation of 3 mouse weight of table
Group | Mouse weight (g) when beginning | At the end of mouse weight (g) |
Normal group | 52.05±1.04 | 54.64±1.23 |
Model group | 65.45±1.15 | 72.76±1.08# |
Cicada fungus buck extract group | 55.18±0.95 | 44.24±1.36*** |
#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 3 it can be seen that the fur of the animal by treatment is more smoother than the fur of model group.At the end of experiment, just
Average weight significant difference (P < 0.05) between normal group and model group, average weight is about when starting for model group mouse
65.45g, average weight is 72.76g at the end of experiment, illustrates high fat diet obesity-induced mice model modeling success;With mould
Type group is compared, and administration group average mice body weight is remarkably decreased (P < 0.001) (table 3).This shows: suitable to obesity mice stomach-filling
Cicada fungus buck extract can significantly slow weight gain.
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 < 0.001).This shows: appropriate to obesity mice stomach-filling
Cicada fungus buck extract total cholesterol, triglycerides and low-density lipoprotein white level in serum can be significantly reduced.
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 cicada fungus buck 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 public affairs
The TruSeq Nano DNA LT Library Prep Kit of department prepares sequencing library.Using MiSeq sequenator carry out 2 ×
The both-end of 300bp is sequenced, and corresponding reagent is 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 Alpha diversity indices of table
Group | Chao1 | Shannon | Simpson |
Normal group | 1520.9811±229.3747 | 6.5011±0.5484 | 0.9633±0.0245 |
Model group | 1365.3829±168.9735# | 5.92±0.4757# | 0.9086±0.0121# |
Cicada fungus buck extract group | 1494.9171±91.2415* | 6.3143±0.3478* | 0.9657±0.0151* |
#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 Simpsonn are aobvious
It writes and increases (P < 0.05), after illustrating that the obesity mice intestinal flora of high fat diet induction gives cicada fungus buck extract for treating, intestines
Road bacterial 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 | Bacteroidetes | Firmicutes |
Normal group | 0.3365±0.1224 | 0.6293±0.1156 |
Model group | 0.2165±0.0867# | 0.7523±0.0896# |
Cicada fungus buck extract group | 0.4665±0.1198*** | 0.4842±0.1352*** |
#P < 0.05,##P < 0.01,###Normal group of P < 0.001vs;*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
It dramatically increases (P < 0.001), Firmicutes relative abundance significantly reduces (P < 0.001).
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, cicada fungus buck extract of the invention have the function of adjusting intestinal bacilli illness.
To sum up it can be seen that, cicada fungus buck extract can not only be clearly helpful for dominant microflora structure in enteron aisle and restore, and have
Help flora diversification, but also significantly treatment and/or preventive effect shown to disease relevant to intestinal microflora,
The deterioration of inflammation can be effectively relieved, improve immune-related every physiopathological index, it is various to can be used for drug, health care product etc.
In field, 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. use of the cicada fungus buck extract in the preparation of preparation treatment and/or prevention intestinal bacilli illness disease and related disease
On the way.
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 cicada fungus buck extract is preparing the use in immunopotentiator
On the way.
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
Cicada fungus buck 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 include cicada fungus buck
Extract and pharmaceutically acceptable carrier.
7. drug according to claim 6, 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.
8. a kind of preparation method of cicada fungus buck extract, feature exist, comprising:
After cicada fungus fructification is crushed, the aqueous alkali measured with 12-16 times ultrasonic extraction 1-2 hours, obtains extracting solution, subtract
PH to 4~5 is adjusted in pressure concentration, and precipitating is precipitated in refrigeration, is taken pellet frozen dry, is obtained cicada fungus buck extract.
9. preparation method according to claim 8, which is characterized in that the aqueous alkali be include NaOH that pH is 9-10
And/or NaHCO3Aqueous solution.
10. the cicada fungus buck extract that preparation method described in a kind of claim 8 or 9 is prepared.
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CN115669932A (en) * | 2021-07-29 | 2023-02-03 | 浙江济公缘药业有限公司 | Composition with function of regulating intestinal flora and preparation method and application thereof |
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