CN110150229A - A kind of production method of diabetes tendon degeneration necrosis disease model rodent - Google Patents
A kind of production method of diabetes tendon degeneration necrosis disease model rodent Download PDFInfo
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Classifications
-
- 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
- A01K67/027—New or modified breeds of vertebrates
-
- 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
- A01K2207/00—Modified animals
- A01K2207/20—Animals treated with compounds which are neither proteins nor nucleic acids
-
- 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
- A01K2207/00—Modified animals
- A01K2207/25—Animals on a special diet
-
- 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
- A01K2227/00—Animals characterised by species
- A01K2227/10—Mammal
- A01K2227/105—Murine
-
- 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
- A01K2267/00—Animals characterised by purpose
- A01K2267/03—Animal model, e.g. for test or diseases
- A01K2267/035—Animal model for multifactorial diseases
- A01K2267/0362—Animal model for lipid/glucose metabolism, e.g. obesity, type-2 diabetes
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Medicines Containing Plant Substances (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of production methods of diabetes tendon degeneration necrosis disease model rodent, comprising giving rodent high lipid food, ambient humidity 70~100%, 28~38 DEG C of environment temperature are kept, continuous 28 days, streptozotocin is injected, diabetes model rodent is obtained；The surface of a wound is manufactured in the foot of the diabetes model rodent, and avoids blood vessel injection staphylococcus aureus 3~8 × 10 along tendon fascia layer in the foot surface of a wound6CFU, after 3 days to obtain the final product.The diabetes tendon degeneration necrosis disease model rodent of method production of the invention can be used for studying the treatment of hot and suffocating card patients with type Ⅰ DM foot gangrene (muscle subcutaneous ulcer) of phlegm wet.
Description
Technical field
The invention belongs to field of medicaments, more particularly to a kind of production of diabetes tendon degeneration necrosis disease animal model
Method.
Background technique
Diabetic foot gangrene (diabetic foot, DF) full name diabetes acromelic gangrene is a kind of different with local nerve
Often and Lower Distal Extremities peripheral vascular disease becomes relevant foot infection, ulcer and (or) deep tissues destruction, be vascular change at lower extremities,
Neuropathy and infection are coefficient as a result, being the most common terminal of diabetic complication development.
National famous physician professor Xi Jiuyi is studied for a long period of time discovery, the tendon necrosis of the Ischemic in diabetic foot gangrene
Type harm is maximum.It is gangrenous (gangrene of finger or toe) that the pathological tissues, treatment and prognosis of this disease are different from ischemic.This disease diseased region begins
Ischemic withered boundary obscission does not occur eventually, is not inconsistent with Chinese medicine " gangrene of finger or toe (ischemic gangrene) " really.Tendon tissue Chinese medicine
It is attributed to " muscle ", therefore why professor intends corresponding names of disease of tcm is " muscle subcutaneous ulcer (tendon degeneration necrosis disease) " (why nine first-class, diabetes flesh
The clinical research of tendon degeneration necrosis disease (muscle subcutaneous ulcer), Shanghai Chinese magazine, the 5th phase in 1996).
Summary of the invention
The present invention provides a kind of production methods of diabetes tendon degeneration necrosis disease model rodent, comprising giving
Rodent high lipid food keeps ambient humidity 70~100%, 28~38 DEG C of environment temperature, and continuous 28 days, injection chain urea helped bacterium
Element obtains diabetes model rodent；The surface of a wound is manufactured in the foot of the diabetes model rodent, and is created in the foot
Blood vessel injection staphylococcus aureus 3~8 × 10 is avoided along tendon fascia layer in face6CFU, after 3 days to obtain the final product.
In a preferred example, the model rodent is rat.
In another preferred example, the ambient humidity is 85%.
In another preferred example, the environment temperature is 33 DEG C.
In another preferred example, the step of production diabetes model rodent includes: to give institute at the 29th day
It states rodent fasting, is injected intraperitoneally streptozotocin, stomach-filling glucose solution after 4h, second of intraperitoneal injection chain urea after 72h
Rhzomorph is helped, selects random blood sugar > 16.7mmol/L rodent after second of injection 72h.
In another preferred example, the size of the foot surface of a wound is 5 × 5mm.
In another preferred example, the dosage of the staphylococcus aureus is 5 × 106cFU。
The details of various aspects of the present invention will be able to detailed description in subsequent chapters.By hereafter and claim
Description, the features of the present invention, purpose and advantage will become apparent from.
Specific embodiment
Muscle subcutaneous ulcer according to Xi Jiuyi professor is theoretical, and it is bad enough that the present invention uses complex factors to establish the hot and suffocating card patients with type Ⅰ DM of phlegm wet
Subcutaneous ulcer (muscle subcutaneous ulcer) animal model, specific modeling thinking are as follows: high fat diet and environment high humidity high temperature cause Organism of Rats phlegm wet hot and suffocating
Card, streptozotocin (Strepozotocin) can selective injury B cells of pancreas, cause meronecrosis, cause blood insulin not
Decline with degree with blood glucose rise.Defect of insulin secretion and insulin resistance are the main pathophysiological bases of diabetes B
And notable feature, lead to the disorder of internal glucose and lipid-metabolism, good diabetes B animal model should have this two side
The characteristic in face.
Studies have shown that high lipid food nursing can cause susceptible type Obesity of Rats and insulin resistance, along with low dose
Multiple injection streptozotocin causes pancreas islet β born of the same parents minor injury, and most animals is made to generate pathoglycemia, forms phlegm wet stagnated heat type II
Patients with type Ⅰ DM rat model.The present invention is first fed with high lipid food, after environment high humidity high fever sifts out phlegm wet stagnated heat type obese rat,
Again with the rat model of low dose of streptozotocin 2 times production of intraperitoneal injection, fat, hyperglycemia is shown with hyperlipidemia, height
Insulinemia and insulin resistance, it is consistent with the feature of diabetes B.By phlegm wet stagnated heat type diabetes B rat foot system
The surface of a wound is made, and is allowed to infect S. aureus L-forms, meets the performance of muscle subcutaneous ulcer morbidity local swelling phase inflammatory reaction, flush, scorching hot, central part
Existing skin lesion necrosis, mostly companion's corruptibility foul gas are separated, purulence liquid is very few after ulceration, and swelling is not easy to decline.The present invention is based on doctors trained in Western medicine
Onset diabetes mechanism and Basic Theories of Chinese Medicine are established the hot and suffocating card patients with type Ⅰ DM foot gangrene of phlegm wet (muscle subcutaneous ulcer) rat model, are made up
The vacancy of Chinese medicine muscle subcutaneous ulcer model, the animal model for meeting theory of traditional Chinese medical science is provided for diabetes Chinese medicine class research.
In turn, the present invention provides a kind of method for building up of hot and suffocating card patients with type Ⅰ DM foot gangrene (muscle subcutaneous ulcer) model of phlegm wet, including
By Wi star rat under hot and humid environment, carries out high glucose and high fat diet 4 weeks, establish phlegm wet stagnated heat type obese rat
Model.Damp heat syndrome of diabetes model was established to rat in 29th day, i.e., whole Rat Fasts be can't help into water 12h, fasting 12h (does not surpass
Cross 15h, normal water), it is injected intraperitoneally (i.p.) streptozotocin (Streptozocin, STZ), stomach-filling glucose is molten after 4h
Liquid, second of i.p. injects STZ after 72h, injects 3 days rear molding venous blood samplings second, and selection measures random through glucometer
Blood glucose > 16.7mmol/L rat.The rat foot surface of a wound is carried out using clipper for surgical use to construct, building foot surface of a wound size about 5 ×
5mm is 1 × 10 with concentration8The S. aureus L-forms suspension of cFU/mL injects 30~80 μ L of surface of a wound muscle, and injection should avoid blood vessel.To hand
After art 3 days, the as hot and suffocating card patients with type Ⅰ DM foot gangrene of phlegm wet (muscle subcutaneous ulcer) animal model.
The method for building up of above-mentioned hot and suffocating card patients with type Ⅰ DM foot gangrene (muscle subcutaneous ulcer) animal model of phlegm wet, specifically includes following step
It is rapid:
(1) Wistar health male rat is chosen, weight is weight 100-120g, pre- to feed 1 week.Free diet drinking-water.
(2) qualified rat is subjected to 40% High-fat diet, while improves air humidity, increase environment temperature (raising
Humid environment (85 ± 15%), and 4h, 33 ± 5 DEG C of temperature are irradiated with heater daily), obese rat is selected after 4 weeks.
(3) the 29th days, obese rat was deprived of food but not water 12h, and row intraperitoneal injection carries out streptozotocin
The injection of (Streptozocin, STZ).STZ is ready-to-use STZ solution, and the sterling of STZ is crystalline powdery object, pH 4.2-
4.5 citrate buffer solution solvent.The STZ solution prepared should be protected from light ice bath, be finished in 10min.
(4) 3 days rear molding venous blood samplings of STZ are injected, selection measures random blood sugar > 16.7mmol/L through glucometer
Rat.Establish phlegm wet card diabetes rat model.
(5) the rat foot surface of a wound is carried out using clipper for surgical use to construct, construct foot surface of a wound size about 5 × 5mm.
It (6) is 1 × 10 with concentration8The S. aureus L-forms suspension of cFU/mL injects the surface of a wound, and injection should avoid blood vessel.Concrete operations are
Staphylococcus aureus-ATCC25923, bacterium colony after scraping passage, 10 times of phosphate buffer (PBS liquid), after gradient dilution, bacterium
Suspension counts.Using spectrophotometer, A value 600, absorbance 1, as 1 × 10 are set8/mL.Each surface of a wound subcutaneous injection should
Concentration 0.1mL PBS liquid.
(7) after performing the operation 3 days, the as hot and suffocating card diabetes of phlegm wet (muscle subcutaneous ulcer) animal model.
Present invention will be further explained below with reference to specific examples.It should be understood that these embodiments are merely to illustrate the present invention
Rather than it limits the scope of the invention.Reagent used in the following example and raw material can be bought by commercial sources to be obtained
?.In the following examples, the experimental methods for specific conditions are not specified, usually according to normal condition or according to proposed by manufacturer
Condition.Unless otherwise defined, it anticipates known to all professional and scientific terms as used herein and one skilled in the art
Justice is identical.In addition, any method similar to or equal to what is recorded and material can be applied to the method for the present invention.Wen Zhong
The preferred implement methods and materials are for illustrative purposes only.
The feature that the features described above or embodiment that the present invention mentions are mentioned can be in any combination.Patent specification is taken off
All features shown can be used in combination with any composition form, and each feature disclosed in specification any can provide phase
The alternative characteristics of same, impartial or similar purpose replace.Therefore except there is special instruction, revealed feature is only impartial or similar
The general example of feature.
Embodiment one
1.1 animals: 60 Wistar male rats, weight 100-120g are purchased from the limited public affairs of Shanghai Si Laike experimental animal
Department, raising are raised in Shanghai Univ. of Traditional Chinese Medicine's Experimental Animal Center, animal certificate number SCXK (Shanghai) 2017-0005, experimental animal
Support credit number SYXK (Shanghai) 2014-0008.Room temperature is adjusted at 20-25 DEG C, and humidity is black in 45% ± 5%, 12h illumination and 12h
Dark situation, standard diet, free water supply are tested after adapting to 7d.
1.2 drugs: the clear muscle side of oriental wormwood be succession residents in Shanghai Xi Jiuyi professor " diabetes muscle subcutaneous ulcer " it is theoretical and according to
It is formed according to " liver governing tendons, the spleen being in charge of the muscles " theory innovation, by the 8 taste Chinese medicinal composition such as oriental wormwood, stringy stonecrop, full side has clearing heat and promoting diuresis, solution
The effect of malicious clear muscle, is clinically used for " muscle subcutaneous ulcer type " patients with diabetic foot gangrene.
1.3 strains: staphylococcus aureus is purchased from Chinese medicine bacterium preservation administrative center, bacterium ATCC25923.
1.4 reagents: chain urea helps mycomycin, is purchased from Sigma Co., USA, article No.: SLBB7534V.Glycosylated hemoglobin
(HbAlC) kit, blood lipids index total cholesterol (TC), triglycerides (TG), low-density lipoprotein (LDL) etc., Nanjing is built up
Bioengineering Research Institute；ELISA kit: tumor necrosis factor α (TNF-α), interleukin-6 (IL-6), c reactive protein
(CRP), glycosylation end products (AGEs), hydroxyproline (Hyp) and myeloperoxidase (MPO), are purchased from the magnificent biology in Wuhan
Engineering Co., Ltd (CUSABIO).
1.5 instruments: a ten thousandth balance BS.124S, German Sartorious instrument company；Desk centrifuge
Centrifuge5702R, German Eppendorf company；High speed freezing centrifuge Centrifuge 5415R, Germany
Eppendorf company；Milli-Q ultrapure water system, Co., Ltd in Millipore；Quick blood sugar radiomete (Johnson & Johnson, the U.S.
The steady bold and unconstrained type blood glucose meter of ONETOUCH)；Microplate reader PowerWave XS, Bio Tek Instrument Ltd., the U.S..
2. method:
2.1 animal packets and diabetes B model foundation
Rat weight is randomly divided into two groups, control group 20, modeling group 50.Control rats are fed with normal diet, make
Mould group feeding environment humid control 85 ± 15%, daily with heater irradiate 4h, 33 ± 5 DEG C of temperature, with 40% high lipid food
Raising 4 weeks, freely drinks syrup (10%).After 4 weeks, there is notable difference in modeling group weight and control group.Fasting 12h (is no more than
15h, normal water), STZ solution 30mg/kg is injected intraperitoneally, injects 50% glucose 3mL of stomach-filling after STZ about 4h；Second after 72h
Secondary injection STZ, method are same as above；It is surveyed on an empty stomach (fasting 7h, normal water) after second of injection 72h, random blood sugar > 16.7mmol/L
Person, that is, diabetes B rat model modeling success, success rate about 80%.Control group injects equivalent volumes buffer, stomach-filling etc.
ML normal saline.
The respective handling of 2.2 muscle subcutaneous ulcer model foundations and control group
Modeling group rat is anaesthetized, by the nearly toe-end cropping in the nearly instep of rat left hind, 75% ethanol disinfection cuts off holostrome skin
Skin, 5 × 5mm of range, and 50 μ L staphylococcus aureus suspension (1 × 10 are injected along tendon fascia layer in the surface of a wound8cFU/mL)。
After performing the operation 3 days, modeling success.After operation, modeling group rat is randomly divided into model group and oriental wormwood Qing Jinfang treatment group is each
20.
Control group group rat is also given similarly anaesthetizes and surgical procedure with above-mentioned modeling group, and it is golden yellow equally to inject 50 μ L
Color staphylococcus suspension (1 × 108cFU/mL)。
3. drug treatment
Oriental wormwood Qing Jinfang treatment group gastric infusion 21g crude drug amount/clear muscle of kg weight oriental wormwood daily to the successful rat of modeling
Side's (clinical 15 times of dosage of adult), successive administration 30 days, once a day.Model group and the isometric physiological saline of control group stomach-filling.
4. observation
4.1 ordinary circumstances:
- as situation observation daily observation rat general status index, including the state of mind, active state, growth body
State, coat, appetite, excrement, urine, and recorded.Weight and the weight of the daily electronic scale scale rat of temperature taking.
4.2 data statistics and analysis data
It is all made of average ± standard deviation expression, statistical software used is SPSS 21.0, and mean value statistical analysis uses single factor test
Method of analysis of variance.P < 0.05 is considered statistically significant difference, and P < 0.01 is considered as that difference is extremely significant.
5. Indexs measure
5.1 blood glucose targets: rat tail point is taken a blood sample weekly, measures random blood sugar value using quick blood sugar radiomete.Experiment knot
Shu Shi is detected glycosylated hemoglobin (HbAlC), colorimetric method, referring to kit specification.
5.2 blood lipids index: using biochemical reagents box, at the end of test experience, TC in each group rat blood serum, TG, and the blood such as LDL
Fat indicators are horizontal, referring to kit specification.
5.4 blood inflammation indexes: using ELISA kit, at the end of test experience, TNF-α in each group rat blood serum,
The inflammatory factors such as IL-6, CRP are horizontal, referring to kit specification.
5.5 metabolic index: ELISA kit, at the end of test experience, glycosylation end products in each group rat blood serum are used
(AGEs), hydroxyproline (Hyp), myeloperoxidase (MPO) are horizontal, referring to ELISA kit specification.
5.6 observation contrast model groups and treatment group's wound healing situation: 1 week, 2 weeks, 3 weeks and 4 after modeling success
The influence of week observation modeling and administration to wound healing, wound healing rate=(initial wound area-residual area)/initial wound
Area × 100%.
6. result
Status index as 6.1-is observed
All rat spirit are good before experiment, are quick on the draw, and active active, coat is submissive pure white glossy, and diet is intended to just
Often, the black hard forming of stool, urine are yellowish.After experiment, the performance of control group Non Apparent Abnormality, weight increasingly increases, the surface of a wound restore compared with
Fastly, 14 days when restore complete substantially.There is wound oedema in 8 rats of muscle subcutaneous ulcer model group, and soft tissue necrosis 14-20 days still relatively tight
Weight, heals 3 substantially after 30 days, does not heal 15, and dead 2.For the clear muscle side's group of oriental wormwood at 14 days, wound was basic compared with model group
Healing 15, does not heal 5, and dead 0.
The White blood cell of 6.2 muscle subcutaneous ulcer model groups and the intervention effect of the clear muscle side of oriental wormwood
Leucocyte index variation (10 before and after 1 each group rat modeling of table^9/ L,)
Note: compared with the control group, P < 0.05 *；Treatment group is compared with model group, #P < 0.05.
Table 1 shows that whole experiment process control rats leucocyte is without significant change.Before modeling, each group rat is white thin
The equal no significant difference of born of the same parents, leucocyte increases compared with the control group for model group and treatment group rat after modeling, (P < 0.05) difference tool
Statistically significant (P < 0.05).
Leucocyte index variation 10 after the treatment of 2 each group rat of table^9/L
Note: compared with the control group, P < 0.05 *；Treatment group is compared with model group, #P < 0.05.
Table 2 shows that whole experiment process control rats leucocyte is without significant change.1 week, 2 weeks, 3 weeks and 4 weeks after treatment
Model group increases compared with control group leucocyte, and difference has statistical significance (P < 0.05).The clear muscle side's group of oriental wormwood is being treated
It is increased compared with control group leucocyte within 1 week afterwards, 2 weeks, difference has statistical significance (P < 0.05).3 weeks after treatment, 4 weeks oriental wormwoods
Clear muscle Fang Zuyu model group compares leucocyte decline, and difference has statistical significance (P < 0.05).
The intervention effect of change of blood sugar caused by 6.3 muscle subcutaneous ulcer model groups and the clear muscle side of oriental wormwood
3 each group rat modeling front and back change of blood sugar of table (mmol/L,)
Note: compared with the control group, P < 0.05 *.
Table 3 shows that whole experiment process control rats blood glucose level is without significant change.Before modeling, each group rat serum
The equal no significant difference of sugar, model group is with blood glucose after the modeling for the treatment of group rat compared with control rats height (P < 0.05), mould after modeling
No significant difference between type group and treatment group.
4 each group rat of table treatment after change of blood sugar (mmol/L,)
Note: compared with the control group, P < 0.05 *；Treatment group is compared with model group, #P < 0.05.
Table 4 shows that whole experiment process control rats blood glucose is without significant change.1 week after treatment, 2 weeks, 3 weeks, 4 weeks moulds
Type group increases compared with control group blood glucose, and difference has statistical significance (P < 0.05).The clear muscle side's group of oriental wormwood after the treatment 1
Week, 2 weeks increase compared with control group blood glucose, and difference has statistical significance (P < 0.05).3 weeks after treatment, 4 weeks clear muscle sides of oriental wormwood
Group blood glucose compared with model group declines, and difference has statistical significance (P < 0.05).
HbAlC and TC caused by 6.4 muscle subcutaneous ulcer model groups, the variation of the blood lipid levels such as TG, LDL and the intervention of the clear muscle side of oriental wormwood
Effect
Table 5 each group rat HbAlC and TC, the variation of TG, LDL
Note: compared with the control group, P < 0.01 * P < 0.05, * *；Treatment group is compared with model group, #P < 0.05.
Table 5 shows that at the end of experiment, muscle subcutaneous ulcer model group HbA1C level significantly increases (P < 0.01) compared with control group, and oriental wormwood is clear
The HbA1C that muscle side's group can be substantially reduced muscle subcutaneous ulcer animal pattern is horizontal (P < 0.05)；Muscle subcutaneous ulcer model group TC level is bright compared with control group
Aobvious to increase (P < 0.05), the TC that the clear muscle side's group of oriental wormwood can be substantially reduced muscle subcutaneous ulcer animal pattern is horizontal (P < 0.05)；Muscle subcutaneous ulcer model
Group TG level is significantly raised (P < 0.05) compared with control group, and the TG that the clear muscle side's group of oriental wormwood can be substantially reduced muscle subcutaneous ulcer animal pattern is horizontal
(P<0.05)；Muscle subcutaneous ulcer model group LDL level is significantly raised (P < 0.05) compared with control group, and the clear muscle side's group of oriental wormwood can be substantially reduced muscle
The LDL of subcutaneous ulcer animal pattern is horizontal (P < 0.05).
The variation of the inflammatory factors level such as TNF-α, IL-6, CRP and the clear muscle side of oriental wormwood in serum caused by 6.5 muscle subcutaneous ulcer model groups
Intervention effect
The variation of the inflammatory factors level such as 6 each group rat blood serum TNF-α of table, IL-6, CRP
Note: compared with the control group, P < 0.01 * P < 0.05, * *；Treatment group is compared with model group, #P < 0.05.
Table 6 is shown, at the end of experiment, TNF-α level is significantly raised (P < 0.05) compared with control group in muscle subcutaneous ulcer model group serum,
The TNF-α that the clear muscle side's group of oriental wormwood can be substantially reduced muscle subcutaneous ulcer animal pattern is horizontal (P < 0.05)；Muscle subcutaneous ulcer model group blood serum IL-6 water
It is flat significantly to increase (P < 0.01) compared with control group, the clear muscle side's group of oriental wormwood can be substantially reduced muscle subcutaneous ulcer animal pattern IL-6 level (P <
0.05)；Muscle subcutaneous ulcer model group CRP level significantly increases (P < 0.01) compared with control group, and the clear muscle side's group of oriental wormwood can be substantially reduced muscle subcutaneous ulcer
The CRP of animal pattern is horizontal (P < 0.05).
6.6 muscle subcutaneous ulcer model group serum glycosylation end products (AGEs), hydroxyproline (Hyp), myeloperoxidase (MPO) etc.
The intervention effect of level variation and the clear muscle side of oriental wormwood
Table 7 each group rat blood serum AGEs, Hyp and MPO variation
Note: compared with the control group, P < 0.01 * P < 0.05, * *；Treatment group is compared with model group, #P < 0.05.
Table 7 is shown, at the end of experiment, the AGEs level of muscle subcutaneous ulcer model group gangrene tissue it is significantly raised compared with control group (P <
0.05), the clear muscle side's group of oriental wormwood can be substantially reduced the AGEs level (P < 0.05) of muscle subcutaneous ulcer animal pattern；Muscle subcutaneous ulcer model group gangrene group
The Hyp level knitted significantly increases (P < 0.01) compared with control group, and the clear muscle side's group of oriental wormwood can be substantially reduced the Hyp of muscle subcutaneous ulcer animal pattern
Horizontal (P < 0.05)；The MPO level of muscle subcutaneous ulcer model group gangrene tissue significantly increases (P < 0.01) compared with control group, the clear muscle side's group of oriental wormwood
The CRP that muscle subcutaneous ulcer animal pattern can be substantially reduced is horizontal (P < 0.05).
The intervention effect of influence and oriental wormwood clear muscle side of the 6.7 muscle subcutaneous ulcer model groups to wound healing rate
8 each group rat wound healing rate of table variation (%,)
Note: compared with the control group, P < 0.01 * P < 0.05, * *；Treatment group is compared with model group, #P < 0.05, ##P < 0.01.
Table 8 shows that the 4th day after surgery, " muscle subcutaneous ulcer " diabetic foot gangrene model modeling was successful, the wound of model group rats
Healing rate is substantially less than control group (P < 0.01).Start to be administered after modeling success, the 1-3 weeks upon administration, model group rats
Wound healing rate is substantially less than control group (P < 0.01)；And give with clearing heat and promoting diuresis, clear muscle effect of detoxifying, clinically it is used for
The treatment group of the name doctor clear muscle side of proved recipe oriental wormwood of " muscle subcutaneous ulcer type " diabetic foot gangrene treatment upon administration the 1st, 2,3 week can be compared with mould
Type group has preferable wound healing rate (P < 0.05), shows diabetic foot gangrene model modeling success of the invention, and meet phlegm
Wet stagnated heat type card type.
7. brief summary: after modeling, muscle subcutaneous ulcer model group is thin in white blood cell count(WBC), random blood sugar value, hemoglobin, blood lipid, inflammation
Intracellular cytokine, serum glycosylation end products (AGEs), hydroxyproline (Hyp), myeloperoxidase (MPO) etc. are and control group
In the presence of the significant difference with statistical significance；In terms of wound healing, the 4th day after operation, the wound healing rate of model group rats
Substantially less than control group.1~3 week after administration, the wound healing rate of model group is substantially less than control group.Pass through the clear muscle side of oriental wormwood
Treatment, the above index have preferable improvement.
Claims (7)
1. a kind of production method of diabetes tendon degeneration necrosis disease model rodent, comprising giving rodent feeding high in fat
Material keeps ambient humidity 70~100%, 28~38 DEG C of environment temperature, continuous 28 days, injects streptozotocin, obtain diabetes mould
Type rodent；The surface of a wound is manufactured in the foot of the diabetes model rodent, and in the foot surface of a wound along tendon fascia
Layer avoids blood vessel injection staphylococcus aureus 3~8 × 106CFU, after 3 days to obtain the final product.
2. the production method of diabetes tendon degeneration necrosis disease model rodent as described in claim 1, the mould
Type rodent is rat.
3. the production method of diabetes tendon degeneration necrosis disease model rodent as claimed in claim 2, the ring
Border humidity is 85%.
4. the production method of diabetes tendon degeneration necrosis disease model rodent as claimed in claim 2, the ring
Border temperature is 33 DEG C.
5. the production method of diabetes tendon degeneration necrosis disease model rodent as claimed in claim 2, the life
The step of producing diabetes model rodent includes: that the rodent fasting was given at the 29th day, and intraperitoneal injection chain urea helps bacterium
Element, stomach-filling glucose solution after 4h, second of intraperitoneal injection streptozotocin after 72h select random blood after injecting 72h second
The rodent of sugar > 16.7mmol/L.
6. the production method of diabetes tendon degeneration necrosis disease model rodent as claimed in claim 2, the foot
The size of portion's surface of a wound is 5 × 5mm.
7. the production method of diabetes tendon degeneration necrosis disease model rodent as claimed in claim 2, the gold
The dosage of staphylococcus aureus is 5 × 106cFU。
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