CN1727005A - Thrombus of deep vein in traumatic limbs and trunk of rat forms the method for building up of animal model - Google Patents
Thrombus of deep vein in traumatic limbs and trunk of rat forms the method for building up of animal model Download PDFInfo
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- CN1727005A CN1727005A CN 200510010795 CN200510010795A CN1727005A CN 1727005 A CN1727005 A CN 1727005A CN 200510010795 CN200510010795 CN 200510010795 CN 200510010795 A CN200510010795 A CN 200510010795A CN 1727005 A CN1727005 A CN 1727005A
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Abstract
The present invention is a kind of method for building up of thrombus of deep vein in traumatic limbs and trunk of rat animal model.It is characterized in that using the orthopaedics intramedullary pin and beat group homemade quantitatively beating unit of device repacking, simulation and the clinical similar wound process of orthopaedics, cause the two lower limb near-end damages in various degree of rat, create a kind of limb deep phlebothrombosis animal model with orthopaedics clinical characters, deep venous thrombosis incidence rate height, and can carry out the detection of different indexs.This invention manufacturing process is simple, the energy wide popularization and application, and the animal dead rate is low; After better simulating the clinical orthopaedics wound, deep venous thrombosis takes place, develops and lapses to process, has application value.
Description
Technical field
The present invention relates to medical modelling technique field, be to set up to have the method for orthopaedics clinical characters thrombus of deep vein in traumatic limbs and trunk animal model, can be research thrombus of deep vein in traumatic limbs and trunk, pulmonary thromboembolism generation, develop, lapse to, explore more effective prevention, diagnosis and treatment method provides an animal model platform.
Background technology
Thrombotic disease is the disease of a class serious harm human health and life, sees a plurality of clinical department patients, and its sickness rate, disability rate and mortality rate are all very high, and hazardness is very big.In western countries, venous thromboembolism (venous thromboembolism, sickness rate VTE) be up to 0.1%, wherein half be deep venous thrombosis (deep venous thrombosis, DVT).U.S. every year because of DVT and pulmonary thromboembolism (pulmonarythromboembolism, PTE) about 650000 examples of number of hospitalized, about 50000～200000 examples of death toll, mortality rate accounts for the 3rd, is only second to tumor and myocardial infarction.The large autopsy data in Europe confirms that the incidence rate of DVT is higher 4 times than arterial thrombus, can find DVT in 40%～60% the cadaver sample, and wherein only 11%～15% case is being made a definite diagnosis before death.The DVT incidence rate of Taiwan and the mortality rate of PTE are about America and Europe's 2/3, the statistics of still unmatchful DVT of China mainland and PTE incidence, and supposition should be similar to the Taiwan incidence.
Along with aged tendency of population, the development of industry, traffic, the high-energy trauma patient increases, and orthopedic injury patient number is in rising trend.In recent years, DVT has become the orthopaedics common complication, and sending out case load obviously increases.DVT often is again the source of PTE, and its risk factor of bringing out PTE is 50%, often causes that the mortality clinical events takes place.Lowxy etc. studies show that the incidence rate of DVT is respectively 50%, 76.5% and 37%～74% behind Fracture of femur, full knee replacement and operation on hip joint.Fracture of tibia 45%～60% is found in external large postmortem, fracture of pelvis 50%～70%, and DVT takes place in the patient of spine and spinal cord injuries 50%～100%; Wounded patient 15%, fracture of pelvis 25%, the concurrent PTE of spinal fracture 14% patient.The embolus that causes PTE mainly comes from lower limb DVT, so DVT and PTE are closely related.
The foundation of animal model is exploration disease reason, pathophysiological change, carries out new therapy, predicts clinical one of the indispensable method that lapses to.In medical research, ideal animal model should possess following characteristics: (1), should reproduce the human diseases that will study, and should be similar with human diseases; (3), animal background complete data, laboratory animal is qualified, life cycle will satisfy the experiment needs; (4), the animal cheapness, the source is sufficient, is convenient to transport.
At the phlebothrombosis animal model following method is arranged aspect setting up both at home and abroad at present: (1) mechanical damage method; (2) ligation method; (3) copper pin energising venous thrombosis method; (4) laser irradiation; (5) rabbit auricular vein thrombosis method; (6) ballon catheter drift vein obstruction method; (7) pico farad in the electrolyte injured blood vessel; (8) place the foreign body method in the blood vessel; (9) method of other venous thrombosis.
Though above modeling method respectively has characteristics, but main is variation and related drugs screening and the therapeutic evaluation that is used to study blood coagulation, fibrinolytic system, conform to not to the utmost with the clinical DVT natural history of orthopaedics reality: the clinical DVT overwhelming majority of (1), orthopaedics is for subacute or chronic, and the formed thrombosis of said method is acute; (2), the clinical DVT of the orthopaedics dark vein of coup injury not nearly all, the equal coup injury blood vessel of said method.(3), the clinical DVT of orthopaedics betides wound or operation back limb brake, long-term bed person mostly, said method does not also meet this condition; (4), more the clinical DVT patient of orthopaedics do not have and use the coagulant history, DVT still takes place in part patient when using anticoagulant.
For further investigation have orthopaedics characteristics deep venous thrombosis and concurrent thus pulmonary thromboembolism generation, develop, lapse to, explore more effective prevention, diagnosis, Therapeutic Method, just be necessary to set up new, more adapt to the animal model of its characteristics.
Summary of the invention
The thrombus of deep vein in traumatic limbs and trunk of rat manufacture method that the present invention proposes, be by using quantitatively beating unit of self-control, simulation and the clinical similar wound process of orthopaedics, cause the two lower limb near-end damages in various degree of rat, cause the limb deep phlebothrombosis to take place, and carry out the detection of corresponding index.
The present invention has following advantage:
(1), manufacturing process is simple, the animal dead rate is low.
(2), limb deep phlebothrombosis incidence rate height.
(3), can better simulate the clinical orthopaedics wound after, clinical and pathological process takes place, develops and lapse to etc. in deep venous thrombosis, can be the research deep venous thrombosis, pulmonary thromboembolism provides an animal model, has application value.
Concrete steps are as follows:
1, animal and grouping
120 Wistar rat male and female are not limit, and after body weight 230g～300g, adaptability fed for two weeks, are divided into A, B, C three at random and organize greatly: the A group is no wound matched group for fracture group, B group for wound does not have the fracture group, C organizes.40 of every big groups are by site difference observing time 3 days observation groups, 5 days observation groups, 7 days observation groups and 9 days observation groups at random again after the modeling.
2, self-control beating unit
Make group device based on intramedullary pin, calibrate can be measured different quality and impact hammer from the highly free-falling different hit energies of 30cm.Impacting the head plane diameter is 1cm, moment blow energy press Ep=mgh and calculate.
3, modeling method
Injection 1% pentobarbital sodium 1ml/kg in the rat abdominal cavity, after anesthesia comes into force, get the ventricumbent position, thigh root is put and is hit on the platform, touch rat femur greater trochanter clearly, use above-mentioned self-control beating unit, impact the rat bilateral thigh near-end outside (greater trochanter to greater trochanter under 1cm) respectively with different strike quality.The external plaster fixation method: with the anhydrous gypsum bandage roller of wide 3cm, to bubble collapse, fixedly the two lower limb of rat are in hip people word bit in the immersion tap water, and the plaster bandage number of plies is 7 layers.
A group: moment hit energy be 4.2 joules, cause Fracture of femur after, row the hip spica cast fix.
B group: moment hit energy be 2.4 joules, do not cause Fracture of femur, row the hip spica cast fix.
The C group: do not impact, the hip spica cast that only goes fixes.
Animal freely drinks water after the modeling, and normal feedstuff is raised, without anticoagulant and antibiotic.
4, observation index
Each organize rat respectively at modeling after 3,5,7,9d when per (put mutually with 10 rats) observes.
(1) gross examination of skeletal muscle
After the modeling 3,5,7,9d observes the two lower limb blood circulations of rat from color and swelling situation, goes intraperitoneal injection 1% pentobarbital sodium 1ml/kg anesthesia again, dissects rat, exposes the latent large vein of bilateral, arteria saphena, observe to have or not thrombosis.Write down two lower limb situations respectively.Put to death rat after getting Pathologic specimen.
(2) pathological observation
Each the time put the latent large vein of the whole piece that cuts, arteria saphena mutually, liquid-solid fixed behind the nearly far-end of labelling with 10% neutral formalin, the paraffin section embedding, HE dyes, the light microscopic microscopy.Whether observe vein and tremulous pulse has the variation of thrombosis and blood vessel wall.The bilateral blood vessel is record respectively.
5, date processing
Use SPSS10.0 version statistical software, the gained result is carried out statistical procedures.Latent large vein incidence of thrombus adopts x
2Check.
The specific embodiment
One, modeling process and observation index
1, animal and grouping
120 Wistar rats, male and female are not limit.After body weight 230g～300g, adaptability fed for two weeks, be divided into A, B, C three at random and organize greatly: the A group is matched group for fracture group, B group for wound group, C group.40 of every big groups, the site difference is divided into 3 days observation groups after the modeling, 5 days observation groups, 7 days observation groups and 9 days observation groups more at random by observing time after the modeling.
2, self-control beating unit
Make group device based on the orthopaedics intramedullary pin, calibrate can use the weight of different quality to impact.The wound energy is pressed Ep=mgh and is calculated.Impacting contact disc diameter is 1cm.
3, modeling method
Injection 1% pentobarbital sodium 1ml/kg in the rat abdominal cavity after anesthesia comes into force, uses above-mentioned self-control beating unit, impacts the rat bilateral thigh near-end outside (greater trochanter of femur to greater trochanter under 1cm) respectively with different strike quality.Blow energy pressed Ep=mgh and calculated moment.
A group: moment hit energy be 4.2 joules (Ep=1.4kg * 10 * 0.3m), the bilateral thigh near-end outside impact once (greater trochanter to greater trochanter under 1cm) respectively, through bone wipe sense or unusual movable confirm to cause Fracture of femur after, row the hip spica cast fix.
B group: moment hit energy be 2.4 joules (Ep=0.8kg * 10 * 0.3m), the bilateral thigh near-end outside impact once (greater trochanter to greater trochanter under 1cm) respectively, do not cause Fracture of femur, and row the hip spica cast fix.
The C group: the hip spica cast that only goes fixes.
Animal freely drinks water after the modeling, and normal feedstuff is raised, without anticoagulant and antibiotic.
4, observation index
Each organize rat respectively at modeling after 3,5,7,9d when per (put mutually with 10 rats) observes.
(1) gross examination of skeletal muscle
After the modeling 3,5,7,9d observes the two lower limb blood circulations of rat from color and swelling situation, goes intraperitoneal injection 1% pentobarbital sodium 1ml/kg anesthesia again, dissects rat, exposes the latent large vein of bilateral, arteria saphena, observe to have or not thrombosis.Write down two lower limb situations respectively.Put to death rat after getting Pathologic specimen.
(2) pathological examination
Each the time put the latent large vein of the whole piece that cuts, arteria saphena mutually, liquid-solid fixed behind the nearly far-end of labelling with 10% neutral formalin, the paraffin section embedding, HE dyes, the light microscopic microscopy.Whether observe vein and tremulous pulse has the variation of thrombosis and blood vessel wall.The bilateral blood vessel is record respectively.
5, statistical procedures
Use SPSS10.0 version statistical software, the gained result is carried out statistical procedures.Latent large vein incidence of thrombus adopts x
2Check.
Two, result
1, laboratory animal situation
A organizes the 3rd day 2 animals because of severe loss of blood death, and A organizes outside 1 animal unknown cause death in the 7th day, and all the other rats all survive.
2, the two lower limb blood circulation perusals of rat
A group: the livid purple swelling of biped occurs early, and development is fast and serious, even sufficient necrosis occurs and come off.1h after the modeling, the rat biped begins swelling, and color is the paleness purple gradually; 1d after the modeling, biped swelling is promptly more obvious, and color becomes darkviolet; 3d after the modeling, biped swelling is serious; 5d after the modeling, 2 rats respectively have 1 foot to come off because of mummification necrosis; 7d after the modeling, other has each 1 foot of 2 rats to come off because of mummification necrosis; 9d after the modeling, other has 1 rat to have 1 foot to come off because of mummification necrosis.
The B group: biped engenders livid purple swelling, progressively development, and the apodia necrosis comes off.12h after the modeling, the rat biped begins swelling; The swelling of 1d biped is obvious after the modeling, the color paleness; 3d after the modeling, biped swelling is more obvious, and color becomes livid purple color: 5d after the modeling, biped swelling is more obvious, does not come off but sufficient necrosis takes place; There are 2 foot swellings to begin to disappear during 7d: to have 5 foot swellings during 9d and disappear.
The C group: the fixing back of hip spica cast 1d, 3d do not see that the livid purple swelling of foot appears in rat; There are 2 foots livid purple swelling to occur when when 2 foots, 7d are arranged during 5d 1 foot, 9d being arranged, do not see that livid purple struttuer all appears in biped.
3, dissect the latent large vein thrombosis of back perusal rat
The A group: see during 3d that the rat inguinal region has obvious hematoma, as seen arteria saphena beats, and latent large vein incidence of thrombus is 25%; As seen arteria saphena beats during 5d, and latent large vein incidence of thrombus is 50%; As seen arteria saphena beats during 7d, sees latent large vein thrombosis, and incidence of thrombus is 80%; As seen arteria saphena beats during 9d, and latent large vein incidence of thrombus is 60%.
The B group: minority rat inguinal region has the limitation hematoma during 3d, and as seen arteria saphena beats, and latent large vein incidence of thrombus is 15%; As seen arteria saphena beats during 5d, and latent large vein incidence of thrombus is 30%; As seen arteria saphena beats during 7d, sees latent large vein thrombosis, and incidence of thrombus is 60%; As seen arteria saphena beats during 9d, and latent large vein incidence of thrombus is 50%.
The C group: as seen arteria saphena beats during 3d, does not see latent large vein thrombosis; As seen arteria saphena beats during 5d, and latent large vein incidence of thrombus is 5%; As seen arteria saphena beats during 7d, sees latent large vein thrombosis, and incidence of thrombus is 10%; As seen arteria saphena beats during 9d, and latent large vein incidence of thrombus is 5%.
Each is organized the latent large vein thrombosis of rat dissection back perusal and sees the following form, through x
2Check, A, B, C compare for three groups, and difference has significant statistical significance (P＜0.01); Further make x
2Cut apart, the A group compares with the B group, and difference has statistical significance (P＜0.05); The A group compares with the C group, and difference has significant statistical significance (P＜0.01); The B group compares with the C group, and difference has significant statistical significance (P＜0.01).
Each is organized rat and dissects the latent large vein thrombosis generation of back perusal (showing with the vein numerical table)
| A organizes (n=74) | B organizes (n=80) | C organizes (n=80) | ||||
| + | - | + | - | + | - | |
| 3d 5d 7d 9d | 4 9 16 12 | 12 9 4 8 | 3 6 12 10 | 17 14 8 10 | 0 1 2 1 | 20 19 18 19 |
(annotate :+no thrombosis generation takes place-is for thrombosis)
4, latent large vein thrombotic disease light microscopy checking of science
The A group: see latent large vein thrombosis during 3d, mostly be the imperfection thrombosis, incidence of thrombus 31%, visible tunica intima of vein damage, vascular endothelial cell strips off, and subendothelial layer is exposed, and minority has the endarterium damage, does not exist but there is thrombosis; Incidence of thrombus 55.6% during 5d, mostly is complete thrombosis, visible tunica intima of vein and vascular endothelial cell damage, and thrombosis shrinks, and with blood vessel wall adhesion, blood vessel wall inflammatory cell infiltration, minority has the endarterium damage, does not see thrombosis; Incidence of thrombus 85% during 7d, mostly is complete thrombosis, visible tunica intima of vein damage, vascular endothelial cell strips off, and subendothelial layer is exposed, has vascular endothelial cell to cover to the thrombosis periphery surface, blood vessel wall inflammatory cell infiltration, minority have the endarterium damage, but intra-arterial is not seen thrombosis; Latent large vein incidence of thrombus 70% during 9d, part is the imperfection thrombosis, and part is complete thrombosis fibrosis, and part new vessels endotheliocyte forms the pipeline that lost shape can be communicated with each other, and makes revascularization, and intra-arterial does not see that thrombosis exists.
B group: see during 3d that minority conceal large vein thrombosis, mostly be the imperfection thrombosis, visible slight tunica intima of vein damage, vascular endothelial cell strips off, and does not see arterial thrombus, conceals large vein incidence of thrombus 10%; Mostly be the imperfection thrombosis during 5d, visible tunica intima of vein and vascular endothelial cell damage, the blood vessel wall inflammatory cell infiltration is not seen arterial thrombus, latent large vein incidence of thrombus 25%; Mostly be complete thrombosis during 7d, visible tunica intima of vein damage, vascular endothelial cell strips off, and subendothelial layer is exposed, the blood vessel wall inflammatory cell infiltration, intra-arterial is not seen thrombosis, latent large vein incidence of thrombus 50%; Part thrombosis fibrosis during 9d, part new vessels endotheliocyte form the pipeline that lost shape can be communicated with each other, and make revascularization, and intra-arterial is not seen thrombosis, latent large vein incidence of thrombus 55%.
The C group: latent large vein thrombosis is rare during 3d, for the imperfection thrombosis, does not see arterial thrombus, latent large vein incidence of thrombus 5%; See during 5d to be the imperfection thrombosis, do not see arterial thrombus, latent large vein incidence of thrombus 15%; See during 7d to be the imperfection thrombosis, visible slight tunica intima of vein damage, tremulous pulse is not seen thrombosis, latent large vein incidence of thrombus 20%; See during 9d to be incomplete thrombosis, do not see the thrombosis fibrosis, part new vessels endotheliocyte forms the pipeline that lost shape can be communicated with each other, and tremulous pulse is not seen thrombosis, latent large vein incidence of thrombus 10%.
The om observation of the latent large vein thrombosis of rat the results are shown in following table, through x
2Check, A, B, C compare for three groups, and difference has significant statistical significance (P＜0.01); Further make x
2Cut apart, the A group compares with the B group, and difference has significant statistical significance (P＜0.01); The A group compares with the C group, and difference has significant statistical significance (P＜0.01); The B group compares with the C group, and difference has significant statistical significance (P＜0.01).
The om observation result (showing) of the latent large vein thrombosis of rat with the vein numerical table
| A organizes (n=74) | B organizes (n=80) | C organizes (n=80) | ||||
| + | - | + | - | + | - | |
| 3d 5d 7d 9d | 5 10 17 14 | 11 8 3 6 | 2 5 11 10 | 18 15 9 10 | 1 3 4 2 | 19 17 16 18 |
(annotate :+no thrombosis generation takes place-is for thrombosis)
Claims (1)
1, a kind of method for building up of thrombus of deep vein in traumatic limbs and trunk of rat model.It is characterized in that using the orthopaedics intramedullary pin and beat group homemade quantitatively beating unit of device repacking, simulation and the clinical similar wound process of orthopaedics, cause the two lower limb near-end damages in various degree of rat, create a kind of limb deep phlebothrombosis animal model manufacture method with orthopaedics clinical characters, and carry out the detection of corresponding index, a situation arises and local respective change to observe deep venous thrombosis, and concrete steps are as follows:
(1), self-control beating unit
Make group device based on the orthopaedics intramedullary pin, calibrate can be measured different quality and impact hammer from the highly free-falling different hit energies of 30cm.Impacting the head plane diameter is 1cm, moment blow energy press Ep=mgh and calculate.
(2), modeling method
Wistar rat male and female are not limit, and body weight 230g～300g, adaptability fed for two weeks, intraperitoneal injection 1% pentobarbital sodium 1ml/kg, after anesthesia comes into force, use above-mentioned self-control beating unit, impact the rat bilateral thigh near-end outside (greater trochanter to greater trochanter under 1cm) respectively with different strike quality.Blow energy pressed Ep=mgh and calculated moment, and was divided into fracture group (4.2 joules), wound according to wound energy difference and do not have three groups of fracture group (2.4 joules) and no wound groups, and all capable hip spica cast fixes.
(3), observation index
Each organize rat respectively at modeling after 3,5,7,9d observes.
1., gross examination of skeletal muscle
After the modeling 3,5,7,9d observes the two lower limb blood circulations of rat from color and swelling situation, goes intraperitoneal injection 1% pentobarbital sodium 1ml/kg anesthesia again, dissects rat, exposes the latent large vein of bilateral, arteria saphena, observe to have or not thrombosis.Write down two lower limb situations respectively.Put to death rat after getting Pathologic specimen.
2., pathological observation
Each the time put the latent large vein of the whole piece that cuts, arteria saphena mutually, liquid-solid fixed behind the nearly far-end of labelling with 10% neutral formalin, the paraffin section embedding, HE dyes, the light microscopic microscopy.Whether observe vein and tremulous pulse has the variation of thrombosis and blood vessel wall.
(4), date processing
Use SPSS10.0 version statistical software, the gained result is carried out statistical procedures.Latent large vein incidence of thrombus adopts χ
2Check.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108742927A (en) * | 2018-06-27 | 2018-11-06 | 新乡医学院 | A kind of thrombus induction experimental provision |
| CN115245143A (en) * | 2022-07-06 | 2022-10-28 | 上海市中西医结合医院 | Construction method of artery occlusive disease animal model |
-
2005
- 2005-05-10 CN CN 200510010795 patent/CN1727005A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108742927A (en) * | 2018-06-27 | 2018-11-06 | 新乡医学院 | A kind of thrombus induction experimental provision |
| CN115245143A (en) * | 2022-07-06 | 2022-10-28 | 上海市中西医结合医院 | Construction method of artery occlusive disease animal model |
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