CN107372323A - The method for building up of cold-dampness diarrhea animal model - Google Patents
The method for building up of cold-dampness diarrhea animal model Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/02—Breeding vertebrates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract
This case is related to a kind of method for building up of cold-dampness diarrhea animal model, and this method includes：Rat is placed in metabolic cage, freely drinks syrup；The next day fasting alternate-day feeding abundance feed and gavage D galactolipins, continuous 10 days；11st 15 day daily to fill frozen water and is placed in low temperature and high relative humidity environment, continuous 5 days；Shigella is injected intraperitoneally within 16th day, is injected again once after 48h, obtains cold-dampness diarrhea animal model.Animal model constructed by the present invention is similar to cold-dampness diarrhea disease clinic disease states, sign, and by repeating to model, prove that its repeatability and stability are good, the Chinese medicine screening suitable for treating cold-dampness diarrhea disease, experimental basis can be provided for the related new drug development of development.
Description
Technical field
The present invention relates to structure pharmaceutical experiment animal model technical field, more particularly to a kind of cold-dampness diarrhea animal model
Method for building up.
Background technology
Diarrhea system causes the dysfunction of the spleen in transport, large intestine conduction mistake department, wetness hyperactivity internal resistance, with big bowel frequency because of being invaded by exogenous pathogen,invasion of exogenous pathogen, or diet internal injury
Number increases, and matter is dilute half congealed or diarrhea with undigested food, very common during (beast) doctor is clinical in very if water sample is the illness that mainly shows.In recent years
Come, domestic and foreign scholars are research diarrhea mechanism, the medicine of exploitation treatment diarrhea, and numerous studies have been carried out to diarrhea animal model.
Its modeling method mainly includes：The diarrhea model of the inductions such as bacterium, virus, Chinese medicine, Western medicine.These models have ignored with mostly
(beast) doctor card type coincide, and modeling factor is single.Therefrom (beast) is cured in card type, and diarrhea is broadly divided into：Diarrhea due to damp-heat, tremble with fear
Wet diarrhea, indigestion diarrhea, splenasthenic diarrhea, kidney-yang deficiency let out multiply with liver depression it is the splenic diarrhea etc..
Ancient book《The hexa-atomic positive big opinion of recording of Plain Questions》Day " people's disease cold-dampness, hair muscle impotence ..., which moistens, to rush down, and blood overflows ", raiser is external
Feel cold-dampness, except the change of body four limbs, can also cause diarrhea；《Ling Shu Miraculous Pivot or Divine Axis all kinds of diseases and ailments begin to give birth to》：" more cold bad foods of then borborygmus are let out, and are eaten
Do not change." propose that experiencing cold-evil then easily causes ingesta to be difficult to digest and assimilate；《Plain Questions is to will very discuss greatly》Day：" sun it
Win, it is burnt under the people that trembles with fear, pass and rushed down to moisten." discuss the generation for arriving cold-dampness diarrhea, the pathogenic cold-damp of external cause duty, the insufficiency of both the spleen and the kindey of internal cause duty.
The actual situation fever and chills that in diarrhea first distinguishes diarrhea are being controlled from wet opinion in Ming Dynasty Dai Sigong states, it is indicated that：The duck person of letting out, it is clear chilly under,
Urine is pure, wet and cold；Ming Dynasty's ZHANG Jie bin proposes that " victory of the sun, it is burnt under cold people that water is evil to be contained ..., therefore passes and moistened for stool
Rush down ", Jiao under cold-evil's invasion is pointed out on the basis of former generation doctor, causes decline of kidney-YANG, it is impossible to which warm spleen soil, then spleen is precise and tiny not
Cloth, the wet interior life of water, cold-dampness aggregation is mingled, therefore diarrhea occurs；Cold-dampness diarrhea is mainly in autumn, and the interpretation of the cause, onset and process of an illness is cold-damp accumulating in the interior, and spleen loses strong
Fortune, is unable to distinguish between the clear and the muddy, and its clinical symptoms shows as thin under rushing down, very such as water sample, aversion to cold, borborygmus of suffering from abdominal pain, and press from both sides poor appetite, gastral cavity abdomen
Vexed swollen, tongue fur is white or white greasy, soft and moderate pulse.It is addicted to drink, strong tea delicious food, pungent etc., long also hurt stomach and intestine are pungent to help fire, delicious food life
Phlegm.Eat and drink immoderately in general or long-term voracious raw and cold, impairment of the spleen sun can be held back, thus it is raw in cold air, epigastric pain occurs, suffers from abdominal pain, let out
Rush down all disease.
It is the basis for carrying out its study mechanism to establish the reliable cold-dampness diarrhea animal model of science.For further further investigated
Diarrhea due to damp-heat essence and seek its effectively treatment Chinese medicine, it is necessary carry out cold-dampness diarrhea animal model replication research.Reason
The card type animal model replication method thought require it is objective, stably, it is reproducible, during the cause of disease, clinical manifestation, curative effect etc. meet (beast)
The guidance of medical knowledge opinion.
The most common are diarrhea due to damp-heat, the more animal models with heat type diarrhea disease of animal model of diarrhea illness in diarrhea
Based on, and the existing animal model of cold-dampness type diarrhea disease its modeling method of showing as has the following disadvantages：(1) model
" interior wet " is made using the method for feeding high glucose and high fat feed mostly, the factor that " irregular diet " is not lost to taste adds
Wherein, the time that this method causes damage to need to taste is longer, and accumulation is difficult in the short time enough " interior wet "；(2) model
The method for using Escherichia coli gavage mostly, Escherichia coli its lead to diarrhea ability after the digestion of stomach and low PH environment subtract significantly
It is weak.
The content of the invention
For technical problem present in prior art, this case provides a kind of method for building up of cold-dampness diarrhea animal model.
To achieve the above object, this case is achieved through the following technical solutions：
A kind of method for building up of cold-dampness diarrhea animal model, wherein, this method includes：Rat is placed in metabolic cage, from
By drinking syrup；The next day fasting alternate-day feeding abundance feed and gavage D- galactolipins, continuous 10 days；Fills ice in 11-15 days daily
Water is placed in low temperature and high relative humidity environment, continuous 5 days；Shigella is injected intraperitoneally within 16th day, is injected again once after 48h, obtains cold-dampness
Diarrhea animal model.
Preferably, the method for building up of described cold-dampness diarrhea animal model, wherein, the syrup concentration is 10%.
Preferably, the method for building up of described cold-dampness diarrhea animal model, wherein, the concentration of the D- galactolipins is
2%, gavage amount is 20ml/kg.
Preferably, the method for building up of described cold-dampness diarrhea animal model, wherein, the frozen water gavage amount is 50ml/
kg。
Preferably, the method for building up of described cold-dampness diarrhea animal model, wherein, the low temperature and high relative humidity environment temperature is
10 ± 2 DEG C, relative humidity is 90 ± 2%.
Preferably, the method for building up of described cold-dampness diarrhea animal model, wherein, the Shigella is sonnei will
Hayes bacterium, culture concentration are 1.5 × 108CFU/ml, the dosage of intraperitoneal injection are same day 1.0ml/kg, and the injection volume after 48h is
0.5ml/kg。
Preferably, the method for building up of described cold-dampness diarrhea animal model, wherein, rat used is SPF level SD rats,
7 week old, body weight 160-180g.
Preferably, the method for building up of described cold-dampness diarrhea animal model, wherein, the symptom of cold-dampness diarrhea includes stool
Thin, afterbody and crissum pollute, poor appetite, ear, nose, coolness of extremities, and tongue fur is in vain and at least one of greasy in vain.
The beneficial effects of the invention are as follows：
Animal model constructed by the present invention is similar to cold-dampness diarrhea disease clinic disease states, sign, and by repeating to build
Mould, it was demonstrated that its repeatability and stability are good, the Chinese medicine screening suitable for treating cold-dampness diarrhea disease, can be that the related new drug of development is ground
Hair provides experimental basis.
Brief description of the drawings
Fig. 1 is the changes of weight tendency chart of rat during the whole modeling of cold-dampness diarrhea.
Embodiment
With reference to embodiment, the present invention is described in further detail, to make those skilled in the art with reference to specification
Word can be implemented according to this.
1.1 animal：SPF level SD rats 30, male and female half and half, 7 week old, body weight 160-180g, it is big to be purchased from Hunan traditional Chinese medicine
Learn Experimental Animal Center, animal quality certification number：SCXK2004-2005.Room temperature adjustment at 20-25 DEG C, humidity 45% ± 5%,
12h illumination and 12h dark surrounds, standard diet, freely feed water.After adapting to 7 days, tested.
1.2 medicine：Hoisting powder proportion of composing：Rhizoma atractylodis 30g, Poria cocos 15g, wrinkled giant hyssop 10g, baked ginger 10g, cassia twig 20g, rhizoma alismatis
10g, dried orange peel 15g, rheum officinale 20g, it is purchased from the big pharmacies of Yantai Rui Kang.Whole medicines are put into 8 times of water first and soak 20min,
To boil by intense fire, to be cooked by slow fire 30min, filtered through gauze；The dregs of a decoction are separately added into 5 times and 3 times of water again, decoct 2 times；Merge 3 times
Filtrate, Buchner funnel filter, and calculate rat dosage, it is clinical equivalent dosage that water-bath, which is concentrated into concentration,.
1.3 strain：Bacillus ceylonensis A, purchased from Shanghai Shi Feng bio tech ltd, bacterium CVCC25931.
1.4 reagent：Yellow Jackets, article No.：P3761, Sigma Co., USA；Absolute ethyl alcohol, analyze pure, Qingzhou Hengsheng
Chemical Co., Ltd.；Glycerine, analyze pure, lot number 100117, Suzhou Long Yue Chemical Co., Ltd.s；Sodium dihydrogen phosphate, analysis is pure,
Tianjin recovery fine chemistry industry research institute；Disodium hydrogen phosphate, analyze pure, Suzhou Chemical Industry Science Co., Ltd of China Airlines Limited；Sodium chloride, point
Analyse pure, lot number 20150410, Chemical Reagent Co., Ltd., Sinopharm Group；Tryptone, lot number 20140608, the extensive and profound in meaning star life in Beijing
Thing technology Co., Ltd；Beef extract, lot number 20131018, the extensive and profound in meaning star biotechnology Co., Ltd in Beijing；Nutrition fine jade
Fat, lot number 20140807, Qingdao GaoKeYuan Hai Bo Bioisystech Co., Ltd；Formaldehyde, analyze pure, Tianjin great Mao chemical reagent
Factory；Rat IL-1 β, IL-10, TNF-α, DAO, D-LA ELISA kit, Beijing strangely-shaped pines bio tech ltd.
1.5 instrument：Electronic balance, Shanghai precision scientific instrument Co., Ltd；AXTG16G table model high speed centrifuges,
Shanghai Zhao Di bio tech ltd；QL-901 whirlpool instrument, its woods Bel's instrument manufacturing Co., Ltd of Haimen City；Table-type low-speed
High-performance refrigerated centrifuge, Changsha Xiang Yi centrifuges Instrument Ltd.；Thermostat water bath, Hengfeng instrument plant of Jintan City；KQ-
300DE type numerical control ultrasonic cleaners, Kunshan Ultrasonic Instruments Co., Ltd.；KDM-1000 type regulating temperature electrothermal covers, Jiangsu outstanding person are auspicious
That Electrical Appliances Co., Ltd；Portable pressure steam sterilizing pot, Guangzhou Kang Mai Medical Devices Co., Ltd.s；Leica slicers, Germany
Leica companies；OlympusDP-71 photomicrographic systems, Japanese Olympus companies；LRH-70 biochemical cultivation cases, Shanghai one are permanent
Scientific instrument Co., Ltd；IMark type ELIASAs, BIO-RAD companies of the U.S.；It is thin to step the auspicious classification of BC5300Vet five animal blood
Born of the same parents' analyzer, Mairui Biological Medical Electronic Co., Ltd., Shenzhen；LG-R-80 blood viscosity instruments, diligent generation Supreme Being science in Beijing
Instrument Ltd.；German Erba XL-640 automatic clinical chemistry analyzers, German Erba companies.
2. method
2.1 animal packets and model establish rat weight, are randomly divided into Normal group, cold-dampness diarrhea model group, lifting
Scattered treatment group, every group 10.Cold-dampness diarrhea model group rats freely drink syrup (10%), odd-numbered day fasting, and even-numbered days are fed with sufficient general
The logical feed and concentration for gavaging D- galactolipins 4ml/200g, D- galactolipin is 2%, common 10d；Then rat is placed in modeling case
(10 ± 2 DEG C, relative humidity 90 ± 2%), while the common 5d of frozen water 10ml/200g are filled daily；Again with bacillus ceylonensis A bacterium solution
(1.5×108CFU/ml (1ml/kg)) is injected intraperitoneally, is injected again 1 time (0.5ml/kg) after 48h.Rats in normal control group is fed with
Normal diet, light water is freely drunk, it is whole in natural environment (humid control 40%-50%, temperature is at 20-25 DEG C)
Gavage, the isometric physiological saline of intraperitoneal injection.Hoisting powder treatment group handles same model group early stage, and modeling terminates rear continuous gavage liter
Drop dissipates (6g/kg) 5 days.Model group and the isometric physiological saline of Normal group gavage.
2.2 cold-dampness diarrhea model property indices detect
2.2.1 after collection of specimens zoopery terminates, 1% yellow Jackets anesthetized rat (30mg/kg) is injected intraperitoneally,
Abdominal aorta is taken a blood sample, and 1ml is gathered in EDTA-K2 anticoagulant tubes, and 1ml is gathered in liquaemin anticoagulant tube, and remaining blood collection exists
(2h is stood in 37 DEG C of insulating boxs, 3500rmp centrifuges serum, saved backup at -20 DEG C) in conventional vacuum heparin tube.Rapidly
1.5cm ileum and 1.5cm colon of the clip since the ileocecus, are placed in soaking at room temperature 15 days in 10% formalin solution.
2.2.2 the general status index of rat, including the state of mind, active state, life are observed in general status observation daily
Long figure, hair, diet desire, excrement, urine, and recorded.Each group rat is quantified according to symptom integral listed in table 1
Clinical symptoms, every symptom is normally long-pending 0 point, and clinical pathological symptoms are more obvious, and integration is higher, is finally united according to the symptom table of quantization
Meter analysis.
2.2.3 body weight and the body weight of the daily electronic scale scale rat of temperature taking, it is front and rear, pretherapy and post-treatment to attack malicious Shigella
With the body temperature of digital clinical thermometer measure rat.
2.2.4 Scours index measure Scours index (DI)=loose stool rate × loose stools level, the loose stools of the rat of loose stool rate=every
The total degree of number/stool.Loose stools level is divided into level Four according to the size of stain scope on filter paper:1 grade is stain diameter<1cm, 2
Level for stain diameter in 1-1.9cm, 3 grades are stain diameter<2-3cm, 4 grades are stain diameter>3cm.
2.2.5 blood routine detection takes the blood of EDTA-K2 anti-freezings to carry out blood routine detection with automatic blood analyzer.
2.2.6 the blood of blood viscosity detection taking heparin sodium anti-freezing carries out blood viscosity detection with blood viscosity instrument.
2.2.7 serum cytokines detection takes the serum preserved at -20 DEG C, thaws, is said according to ELISA kit at room temperature
Bright progress IL-1 β, TNF-α, IL-10, DAO, D-LA measure.
2.3 data statistics represent that statistical software used is SPSS21.0 with analyze data using average ± standard deviation,
Average statistical analysis one-way analysis of variance method.P<0.05 is considered as statistically significant difference, P<0.01 is considered as difference
It is extremely notable.
3. result
3.1 general status index observings
All rat spirit are good before experiment, are quick on the draw, and active active, hair is submissive pure white glossy, and diet is intended to just
Often, black shaping, the urine firmly of defecating are yellowish.After experiment, Normal group Non Apparent Abnormality performance, body weight increasingly increases.Model group,
In the high glucose and high fat stage, preceding 1-5 days excrement is soft, full mellow and full, one day amount of drinking water of fasting, urine volume increase, and urine is yellowish；To food
One day urine volume of gavage D- galactolipins is relatively fewer, and excrement is partially white, has partial rat to have loose bowels just at the 6-8 days, then defecate by
Band of gradually becoming a cadre is yellow, and there is bleeding in nostril, and other are without significant change.In the low temperature and high relative humidity stage, early stage, the basic shaping of stool sticked greasy hair
Huang, later stage have part phenomenon of having loose bowels occur, into after low temperature bin, anus temperature decrease to some degree, and lassitude (P<0.01),
Body is unfolded, lazy move or motionless (P<0.01), hair is turned to be yellow (P in a jumble<0.01), lose the appetite (P<0.01) under, scrotum is loose
Hang down；After removal low temperature bin enters natural environment, body temperature is just started and has been slightly below normal body temperature, the state of mind, active state are bright later
It is aobvious to recover.After attacking malicious Shigella 5h, the highly depressed, photophobia of spirit, shiver with cold, roll up, flock together (P<0.01), eyelid is red and swollen and divides
Secretions, the even useless (P absolutely of anorexia<0.01), drink is intended to decline (P<0.01), there is diarrhea symptom in 5-8h, and hair is mixed and disorderly
Turn to be yellow (P<0.01), body temperature rise (P<0.01), the red and swollen even expansion (P of most of anus<0.01), diarrhea with loose stool or it is glutinous it is greasy,
Color Huang stench, partly there is mucopurulent bloody stool (P<0.01), crissum and the turbid dirty adhesion yellow loose stools (P of root of the tail<0.01), partly go out
Existing white and greasy tongue coating.The same model group of Hoisting powder treatment group modeling early stage symptom, spirit improvement (P after 24h is administered<0.05), dietary amount, work
Momentum gradually increases (P<0.05), symptom substantially mitigates after 2 days, temperature recovery to normal level, hair is gradually glossy (P<
0.05), diarrhea symptom mitigation (P<0.05), symptom returns to normal after 4 days.
3.2 Scours index
The cold-dampness diarrhea rat model of table 1 Scours index (N=10)
| Group | First day | Second day | 3rd day | 4th day | 5th day |
| Normal group | 0 | 0 | 0 | 0 | 0 |
| Model group | 1.51±0.27* | 1.57±0.26* | 1.46±0.26* | 1.01±0.16* | 0.91±0.23* |
| Hoisting powder group | 1.22±0.21* | 1.07±0.20* | 0.87±0.16*# | 0.45±0.21*# | 0.25±0.15*# |
Note：Compared with Normal group*P<0.05；Compared with model group#P<0.05。
Table 1 shows that model group was compared with Normal group in the 1-5 days, and each Scours index of model group is noticeably greater than normal
Control group (P<0.05)；Hoisting powder group is compared with Normal group within the 1-4 days, and each Scours index of Hoisting powder group is right better than normal
According to group (P<0.05)；Hoisting powder group is compared with Normal group within 5th day, no significant difference (P>0.05)；Rises for 2-5 days
Drop dissipates group and compared with model group, and each Scours index of Hoisting powder group is significantly less than model group (P<0.05).
3.2 body weights change
The changes of weight situation of rat during the whole modelings of Fig. 1
Fig. 1 is shown, during whole modeling, the rat body weight of Normal group is gradually increasing.Model group and root tuber of aromatic turmeric group
The 1-10 days high glucose and high fat stages, feed sufficient feed and fill one day body weight rising of oil, one day Body weight loss of fasting, but always
Trend body weight is gradually increasing；Low temperature and high relative humidity stage body weight is gradually reduced；Body weight is gradually increasing after medicine feed, and lift in Hoisting powder group
Degree is more than model group.
3.3 anus temperature change
The each group rat modeling of table 2 is front and rear, after treatment anus temperature change (N=10)
| Group | Before modeling | After modeling | After treatment |
| Normal group | 36.78±0.25 | 36.95±0.24 | 36.82±0.31 |
| Model group | 36.73±0.29 | 32.45±0.32**## | 35.53±0.27*&# |
| Hoisting powder group | 36.61±0.32 | 32.78±0.25**## | 36.71±0.21&&Δ |
Note：Compared with before this group of modeling,*P<0.05,**P<0.01；After treatment compared with after this group of modeling,&P<0.05,&&P
<0.01.The phase same time, compared with Normal group,#P<0.05,##P<0.01；Pulvis curcumae group compared with model group,ΔP<0.05。
Table 2 shows that rat temperature is before and after modeling, there occurs great changes before and after treatment：Whole experiment process is normally right
According to group rat anus temperature normally without significant change.Before modeling, each group rat temperature is normal, model group and Hoisting powder group after modeling
Body temperature pole significantly reduces (P<0.01)；Body temperature significantly raises (P after Hoisting powder group gavages Hoisting powder<0.01), basically reach normally,
And model group does not have the obvious (P of Hoisting powder group rise<0.05).
3.4 routine blood indexes
The each group rat of table 3 routine blood indexes (N=10)
Note：The different alphabetical person's significant difference (P of same column shoulder mark<0.05)
Table 3 shows that leukocyte count, neutrophil cell number, red blood cell number, packed cell volume, the hemoglobin in blood routine are dense
Degree is that model group is noticeably greater than Normal group (P<0.05), and Hoisting powder group is significantly less than model group (P<0.05), explanation
Cold-dampness diarrhea animal pattern body has inflammatory reaction and reaction of catching cold, and is significantly alleviated after Hoisting powder treatment.
3.5 blood viscosity indexs
The each group rat of table 4 blood viscosity (N=10)
| Group | 200s-1(mpa.s) | 30s-1(mpa.s) | 5s-1(mpa.s) | 1s-1(mpa.s) |
| Normal group | 3.52±0.34 | 4.61±0.31 | 9.53±1.24 | 10.57±1.68 |
| Model group | 5.21±0.46** | 7.17±0.64** | 14.58±2.85** | 15.41±0.39** |
| Hoisting powder group | 3.15±0.39## | 5.02±0.89## | 10.41±3.11## | 10.22±1.54## |
Note：Model group compared with Normal group,**P<0.01；Hoisting powder group compared with model group,##P<0.01.Table 4 is aobvious
Show, 200s-1、30s-1、5s-1、1s-1Under several shear rates, the blood viscosity of model group is all higher than Normal group (P<0.01)；
200s-1、30s-1、5s-1、1s-1Under four shear rates, Hoisting powder group pole is significantly less than model group (P<0.01), 5s-1Shear rate
Lower Hoisting powder group is significantly less than model group (P<0.05).Illustrate the blood viscosity rise of cold-dampness diarrhea animal pattern, Hoisting powder has
Significantly alleviate the blood viscosity effect of cold-dampness diarrhea animal pattern.
3.6 serum cytokines indexs
The each group rat of table 5 serum cytokines (N=10)
Note：Model group compared with Normal group,**P<0.01；Hoisting powder group compared with model group,#P<0.05,##P<
0.01。
Table 5 shows that the IL-1 β poles of model group are significantly higher than Normal group (P<0.01), Hoisting powder group pole is substantially less than
Model group (P<0.01)；The IL-10 poles of model group are substantially less than Normal group (P<0.01), Hoisting powder group is significantly higher than model
Group (P<0.05)；The TNF-α pole of model group is significantly higher than Normal group (P<0.01), pulvis curcumae group is substantially less than model group (P
<0.05).Illustrate the inflammatory reaction rise of cold-dampness diarrhea animal pattern, the inflammatory reaction decline after pulvis curcumae is treated.Model group
DAO poles are significantly higher than Normal group (P<0.01), Hoisting powder group is less than model group (P<0.01)；The D-L of model group is higher than just
Normal control group (P<0.01), Hoisting powder group is less than model group (P<0.05).Illustrate that the gut barrier of cold-dampness diarrhea animal pattern is received
To destruction, it is restored after Hoisting powder is treated.
Although embodiment of the present invention is disclosed as above, it is not restricted in specification and embodiment listed
With it can be applied to various suitable the field of the invention completely, can be easily for those skilled in the art
Other modification is realized, therefore under the universal limited without departing substantially from claim and equivalency range, it is of the invention and unlimited
In specific details and shown here as the legend with description.
Claims (8)
1. a kind of method for building up of cold-dampness diarrhea animal model, it is characterised in that this method includes：Rat is placed in metabolic cage
In, freely drink syrup；The next day fasting alternate-day feeding abundance feed and gavage D- galactolipins, continuous 10 days；The the one 11-15 days daily
Fill frozen water and be placed in low temperature and high relative humidity environment, continuous 5 days；Shigella is injected intraperitoneally within 16th day, injects once, obtains again after 48h
Cold-dampness diarrhea animal model.
2. the method for building up of cold-dampness diarrhea animal model as claimed in claim 1, it is characterised in that the syrup concentration is
10%.
3. the method for building up of cold-dampness diarrhea animal model as claimed in claim 1, it is characterised in that the D- galactolipins it is dense
Spend for 2%, gavage amount is 20ml/kg.
4. the method for building up of cold-dampness diarrhea animal model as claimed in claim 1, it is characterised in that the frozen water gavage amount is
50ml/kg。
5. the method for building up of cold-dampness diarrhea animal model as claimed in claim 1, it is characterised in that the low temperature and high relative humidity environment
Temperature is 10 ± 2 DEG C, and relative humidity is 90 ± 2%.
6. the method for building up of cold-dampness diarrhea animal model as claimed in claim 1, it is characterised in that the Shigella is Song
Nei Shi Shigellas, culture concentration are 1.5 × 108CFU/ml, the dosage of intraperitoneal injection are same day 1.0ml/kg, the note after 48h
The amount of penetrating is 0.5ml/kg.
7. the method for building up of cold-dampness diarrhea animal model as claimed in claim 1, it is characterised in that rat used is SPF levels
SD rats, 7 week old, body weight 160-180g.
8. the method for building up of cold-dampness diarrhea animal model according to claim 1, it is characterised in that the symptom of cold-dampness diarrhea
Thin including stool, afterbody and crissum pollute, poor appetite, ear, nose, coolness of extremities, and tongue fur is in vain and at least one of greasy in vain.
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