CN104116572A - Method for building rat spinal cord injury model - Google Patents
Method for building rat spinal cord injury model Download PDFInfo
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- CN104116572A CN104116572A CN201410408369.6A CN201410408369A CN104116572A CN 104116572 A CN104116572 A CN 104116572A CN 201410408369 A CN201410408369 A CN 201410408369A CN 104116572 A CN104116572 A CN 104116572A
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- spinal cord
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Abstract
The invention relates to a new method for building a rat spinal cord injury model. The method mainly solves the problems that in an existing model, operation is complex, the repeatability is poor, the amount of bleeding of a rat is large, the infection death rate is high, numerous complications are caused and the survival time of the rat is short. According to the technical scheme, unilateral lumbar cord dividing injury is made at the accurate position of the spinal cord of the rat through a homemade syringe needle without opening a medullary space, the spinal cord injury model is built according to the new method, and medicine and preparations can be injected at the same time of making the injury to interfere in the injury and the regeneration and repair process after the injury.
Description
Technical field
The present invention relates to the method for building up of spinal cord injury model, be specifically related to a kind of Model of Rat Spinal Cord Injury method for building up.
Background technology
Spinal cord injury is the disease that a kind of disability rate is very high, is unsolved problem so far clinically, can cause the sensation of movement dysfunction that cannot recover.Along with the fast development of social every profession and trade, the sickness rate of spinal cord injury also increases thereupon.Therefore, the research of spinal cord injury and concern are still the problem of domestic and international joint exploration.And the research that spinal cord injury model is spinal cord injury provides scientific research basis.Desirable spinal cord injury model should possess following condition.One, want nervous physiology that can the damage of reaction experiment animal spinal cord, the situation of motor behavior.Two, can provide the animal model consistent or similar with clinical spinal cord injury, there is good clinical correlation.Three, model will have reproducibility.Research spinal cord injury needs a large amount of laboratory animals, and this just needs the standardization of modeling.
The common spinal cord injury model of prior art has following several:
1, contusion type spinal cord injury model
Spinal cord dorsal part that Allen designs first hits model, utilizes the counterweight of Different Weight vertically to fall from differing heights along sleeve pipe to hit rat spinal cord dorsal part to cause damage in various degree.The weak point of this model: the repeatability that operates is poor, and influence factor is a lot, and phase missionary society is very large often in the damage causing as same potential energy, and the shape and the area that at strike moment impact hammer, contact with spinal cord differ.
2, clamp type spinal cord injury
This model is the research needs that meet the multiple type of impairments such as acute spinal cord extruding.Concrete operations flow process: SD rat vertebral plate is opened in operation, the spinal cord segment with aneurysm clamp clamp corresponding to T10.The deficiency of this model: this model is larger to the damage of rat in setting up, and need to sting the vertebral plate except T10, makes fully to expose corresponding to the spinal cord segment of T10, and rat infection probability is larger, easily infects dead.And clamp is different to the dynamics of spinal cord injury, easily causes rat lower limb panplegia.Rat postoperative care is caused to a lot of inconvenience.
3, cutting-type spinal cord injury model
Flow process: open vertebral plate, expose corresponding spinal cord, use the complete cross-section or half cross-section spinal cord of sharp keen iris blade or microscissors, or one section of spinal cord of excision.The deficiency of the method: operating procedure is more complicated, larger to the damage of rat, and more neat to the transverse section of spinal cord with blade, there is different with clinical practice case.
4, ischemia of spinal cord and re-perfusion model
Be through femoral arteriography or directly close the method analogue formation of descending aorta.The deficiency of the method: operation has also caused the damage of other histoorgans of perfusion district when interruption artery causes ischemia, has affected ethological observation.
Summary of the invention
The present invention overcomes the deficiencies in the prior art, technical problem to be solved is complicated operation in prior art spinal cord injury model, poor repeatability, the amount of bleeding of rat is large, infect the problems such as death is high, complication is many, the time-to-live is short, and then a kind of simple to operate, reproducible, significant Model of Rat Spinal Cord Injury method for building up of dysfunction is provided.
For solving the problems of the technologies described above, the technical solution adopted in the present invention is: Model of Rat Spinal Cord Injury method for building up, carries out in accordance with the following steps:
(1) rat is weighed, 1% pentobarbital sodium inj, carries out lumbar injection by every 100g body weight 0.4ml;
(2) after rat anesthesia, back preserved skin, sterilization skin;
(3) determine T
9-T
12position, SD In Rat Lumbar section spinous process is outstanding, and the hypsokinesis of breast section spinous process, and waist section spinous process leans forward, and according to this sign, is easy to accurate location.At T
9-T
12directly over otch, do one 2 ~ 3cm skin incision, separating muscle and fascia, expose T
9~ T
12spinous process and vertebral plate;
(4) use tissue shear and mosquito forceps by T
9-T
10muscle and the ligament in spinous process left side are peeled off thorough exposure T
9-T
10spinous process and transverse process, determine T
9-T
10gap;
(5) avoid spinal cord center, the side of slightly taking back, by the 1ml syringe needle polishing from T
9-T
10gap is vertically inserted pulp cavity and is entered correspondent section spinal cord.Owing to being negative pressure in pulp cavity, intervertebral space is soft tissue, so does not need to exert oneself very much during inserting needle, and syringe needle is easy to enter pulp cavity and spinal cord.When syringe needle enters after spinal cord, rat can violently shake, pulls and pushes syringe needle vertically, and feeling has larger activity space, shows that syringe needle is in pulp cavity;
Extract syringe needle, the broken broach syringe needle of bending is vertically inserted from entry point, during inserting needle, the needle point of bending is towards left side.The syringe needle that slightly swings after broken broach syringe needle enters pulp cavity, attention will definitely be avoided crooked syringe needle to surpass spinous process center line and enter right Hemimyelia, only destroys thus left side spinal cord.While carrying out this operation, left side (damage side) hind leg has a convulsion, and syringe needle is also still extracted with vertical angle in light and slow ground, and now spinal cord half amputation injury model establishment step completes substantially;
(6) art finishes suture muscles and skin.After rat is clear-headed in dry, peace and quiet, under natural light condition, normal diet is fed, and regularly uses povidone iodine cleaning-sterilizing.
The above-mentioned method of setting up spinal cord injury model also can operate right side spinal cord, and operating condition is corresponding with the above-mentioned operation that left side spinal cord is carried out.
The syringe needle of described staight needle is for putting down without sharp syringe needle; Can be used in the entry point of determining broken broach.
Described staight needle preferably polishes the syringe needle of syringe to make, and the staight needle obtaining, can also be for damaging, also promote the preparation of regenerating after damage except determining brokenly the entry point of broach to interventions such as spinal cord injury position injection neurotrophic factor or neural stem cell.
The syringe needle of described broken broach is right angle syringe needle, described broken broach when vertical lower pin, the free end level of described right angle syringe needle.
Described broken broach be preferably by the needle slope of syringe backward 90-degree bent make.
Material required for the present invention and apparatus are as follows:
Laboratory animal
Healthy male rat, body weight 180-200 gram;
Experiment apparatus
No. 11 knife blades, needle forceps, mosquito forceps, vascular forceps, surgical forceps, tissue shear, suture needle, staight needle (homemade 1 milliliter of syringe needle: polishes 1 milliliter of syringe needle to make, as shown in Figure 1), broken broach (is made 1 milliliter of syringe needle, as shown in Figure 2) to needle slope dorsal flexion 90 degree.
Compared with prior art the present invention has following beneficial effect.
The spinal cord injury model causing with syringe needle is easy and simple to handle, damages littlely, and repeatability is high, and dysfunction is remarkable, and stimulates or promote medicine or the preparation of spinal cord regeneration that good condition and environment is provided for research; For exploring the molecular mechanisms of action of clinical medicine, provide experiment basis.Meanwhile, because animal pattern postoperative infection probability is low, high, the favorable repeatability of few intercurrent disease, survival rate, there is the good prospect of promoting the use of.
Accompanying drawing explanation
Fig. 1 is the structural representation of staight needle of the present invention.
Fig. 2 is the structural representation of the broken broach of the present invention.
Fig. 3 is Spinal Cord Injury in Rats position schematic diagram.
Fig. 4 is Spinal Cord Injury in Rats position spinal cord vertical section schematic diagram.
Fig. 5 is Spinal Cord Injury in Rats position spinal cord transection face schematic diagram.
The specific embodiment
Below in conjunction with specific embodiment, the invention will be further described.
Modeling method:
(1) rat is weighed, 1% pentobarbital sodium inj, carries out lumbar injection by every 100g body weight 0.4ml.
(2) after rat anesthesia, back preserved skin, sterilization skin.
(3) determine T9-T12 position, SD In Rat Lumbar section spinous process is outstanding, and the hypsokinesis of breast section spinous process, and waist section spinous process leans forward, and according to this sign, is easy to accurate location.Otch directly over T9-T12, does one 2 ~ 3cm skin incision, and separating muscle and fascia expose T9 ~ T12 spinous process and vertebral plate.
(4) with tissue shear and mosquito forceps, the muscle in T9-T10 spinous process left side and ligament are peeled off to spinous process and the transverse process of thorough exposure T9-T10 as shown in Figure 3, determined T9-T10 gap.
(5) avoid as shown in Figure 4 spinal cord center, the side of slightly taking back, vertically inserts the 1ml syringe needle polishing from T9T10 gap pulp cavity and enters correspondent section spinal cord.Owing to being negative pressure in pulp cavity, intervertebral space is soft tissue, so does not need to exert oneself very much during inserting needle, and syringe needle is easy to enter pulp cavity and spinal cord.When syringe needle enters after spinal cord, rat can violently shake, pulls and pushes syringe needle vertically, and feeling has larger activity space, shows that syringe needle is in pulp cavity.
Extract syringe needle, as shown in Figure 5 the broken broach syringe needle of bending is vertically inserted from entry point, during inserting needle, the needle point of bending is towards left side.The syringe needle that slightly swings after syringe needle enters pulp cavity, enters right Hemimyelia but definitely avoid crooked syringe needle to surpass spinous process center line, only destroys thus left side spinal cord.While carrying out this operation, left side (damage side) hind leg has a convulsion, and syringe needle is also still extracted with vertical angle in light and slow ground, and now spinal cord half amputation injury model establishment step completes substantially.
(6) art finishes suture muscles and skin.After rat is clear-headed in dry, peace and quiet, under natural light condition, normal diet is fed, and regularly uses povidone iodine cleaning-sterilizing.
Experimental result is as follows.
Rat behavior is learned and is observed
Normal rat four feet contact to earth, can be upright.Spinal cord after clear-headed half is red and swollen from disconnected art rat model postoperative 3-5 day operative incision place, recovers normal after a few days.Contrast with normal rat, acroparalysis, the walking that mops floor behind animal pattern left side, and can not be upright.But rat can carry out normal daily routines, diet and excretion are all normal, and post-operative complication is few, and the time-to-live is longer.Offside hind leg is movable normal.
Rat hindlimb is in grip (motor function) and burning pain (sensory function) reaction of different time
We are by carrying out can finding out in grip and hot plate reaction test from disconnected postoperative rat to spinal cord half, and sensation and the motor capacity of two hind legs of (as table 1, table 2) preoperative rat are all normal.After operation, As time goes on, the sensation of movement of Ipsilateral hind leg and reflection function are all without recovering, and strong rear flank limb motor function is normal, after hot plate reaction is disappeared, gradually recovers to some extent.This explanation half has caused neurological disorder from disconnected art to rat.
Myeloid tissue's morphological observation
Perusal: postoperative 24h rat wound swelling, open after pulp cavity visible waist and expand left side, place spinal cord and significantly rupture, hypermyelohemia swelling.Skin wound healing after 1 week, swelling disappears.Micro-Microscopic observation: the damage zone visible stringer spinal cord fibrous fracture of postoperative rat spinal cord.
Above result can be found out and the invention has the advantages that:
(1) the present invention uses that 1ml syringe needle easily operates first watch, damage position is accurate, and half from completely disconnected, and the spinal cord injury broken ends of fractured bone is closer to clinical practice situation.
(2) the present invention uses syringe needle to damage, little to the damage of rat, and the wound spontaneous recovery time is short, and the infection probability that spinal cord is caused is little, and amount of bleeding is few, less demanding to experiment equipment, and operation is relatively easy, and the sensation of movement obstacle causing is clear and definite.
(3) feature of maximum of the present invention be the syringe that can connect by injection needle at the position injectable drug of damage, neurotrophic factor or neural stem cell etc. are intervened damage, and are promoted the preparation of regeneration after damage.Meanwhile, by spinal cord half, from disconnected art, can observe and inquire into neurotransmitter, neurotrophic factor, medicine and neural stem cell etc. to the impact of spinal cord injury and regenerative process and molecular mechanism thereof.Because model of the present invention can contrast with strong side and can utilize syringe needle injection in spinal cavity, therefore there is unique superiority.
The spinal cord injury model causing with syringe needle is easy and simple to handle, damages littlely, and repeatability is high, and dysfunction is remarkable, and stimulates or promote medicine or the preparation of spinal cord regeneration that good condition and environment is provided for research; For exploring the molecular mechanisms of action of clinical medicine, provide experiment basis.Meanwhile, because animal pattern postoperative infection probability is low, high, the favorable repeatability of few intercurrent disease, survival rate, there is the good prospect of promoting the use of.
The present invention can summarize with other the concrete form without prejudice to spirit of the present invention or principal character.Therefore, no matter from which point, above-mentioned embodiment of the present invention all can only think explanation of the present invention can not limit invention, claims have been pointed out scope of the present invention, and scope of the present invention is not pointed out in above-mentioned explanation, therefore,, in the implication suitable with claims of the present invention and any variation in scope, all should think to be included in the scope of claims.
Claims (9)
1. Model of Rat Spinal Cord Injury method for building up, is characterized in that in accordance with the following steps:
(1) rat is weighed, by every 100g body weight, with 0.4ml pentobarbital sodium inj, carry out lumbar injection;
(2) after rat anesthesia, back preserved skin, sterilization skin;
(3) determine T
9-T
12position, at T
9-T
12directly over otch, do a 2-3cm skin incision, separating muscle and fascia, expose T
9-T
12spinous process and vertebral plate;
(4) by T
9t
10muscle and the ligament on spinous process left side or right side are peeled off thorough exposure T
9-T
10spinous process and transverse process, determine T
9-T
10gap;
(5) avoid spinal cord center, the side of slightly taking back or right side, by staight needle from T
9-T
10gap is vertically inserted pulp cavity and is entered correspondent section spinal cord, extracts syringe needle, forms entry point; The syringe needle of broken broach is vertically inserted from entry point, and entry point is when left side, and during inserting needle, crooked needle point is towards left side; Entry point is when right side, and during inserting needle, crooked needle point is towards right side; The syringe needle that slightly swings after syringe needle enters pulp cavity, enters another Hemimyelia but definitely avoid crooked syringe needle to surpass spinous process center line, only destroys thus left side or right side spinal cord; Also still with vertical angle, extract brokenly the syringe needle of broach then light and slowly;
(6) suture muscles and skin, after rat is clear-headed, under dry, peace and quiet, natural light condition, normal diet is fed, and cleaned at regular intervals sterilization.
2. Model of Rat Spinal Cord Injury method for building up according to claim 1, is characterized in that: the concentration of described pentobarbital sodium inj is 1%.
3. Model of Rat Spinal Cord Injury method for building up according to claim 1, is characterized in that: the syringe needle of described staight needle is for putting down without sharp syringe needle.
4. Model of Rat Spinal Cord Injury method for building up according to claim 3, is characterized in that: described staight needle is for determining the entry point of broken broach.
5. Model of Rat Spinal Cord Injury method for building up according to claim 3, is characterized in that: described staight needle is for to polish the syringe needle of syringe to make.
6. Model of Rat Spinal Cord Injury method for building up according to claim 5, is characterized in that: described staight needle, also can be for injectable drug in spinal cavity for determining the entry point of broken broach.
7. Model of Rat Spinal Cord Injury method for building up according to claim 1, is characterized in that: the syringe needle of described broken broach is right angle syringe needle, described broken broach when vertical lower pin, the free end level of described right angle syringe needle.
8. Model of Rat Spinal Cord Injury method for building up according to claim 7, is characterized in that: described broken broach for by the needle slope of syringe backward 90-degree bent make.
9. according to the Model of Rat Spinal Cord Injury method for building up described in claim 5 or 8, it is characterized in that: described syringe is common 1ml disposable syringe.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410408369.6A CN104116572A (en) | 2014-08-19 | 2014-08-19 | Method for building rat spinal cord injury model |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410408369.6A CN104116572A (en) | 2014-08-19 | 2014-08-19 | Method for building rat spinal cord injury model |
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| Publication Number | Publication Date |
|---|---|
| CN104116572A true CN104116572A (en) | 2014-10-29 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410408369.6A Pending CN104116572A (en) | 2014-08-19 | 2014-08-19 | Method for building rat spinal cord injury model |
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2357710Y (en) * | 1999-02-24 | 2000-01-12 | 郑利平 | Disposable lachrymal irrigator |
| CN2560367Y (en) * | 2001-08-30 | 2003-07-16 | 夏勤 | Needle head for dropping medicine suitable for throat and trachea |
| CN201260729Y (en) * | 2008-06-30 | 2009-06-24 | 于青云 | Novel lacrimal passage rinser |
| CN201929962U (en) * | 2010-12-17 | 2011-08-17 | 邱晨 | Blood collection injector |
| CN202776492U (en) * | 2012-04-11 | 2013-03-13 | 上海中医药大学附属曙光医院 | Tail unit curved needle |
| CN103462720A (en) * | 2013-07-24 | 2013-12-25 | 董延庆 | Syringe for veterinarian |
-
2014
- 2014-08-19 CN CN201410408369.6A patent/CN104116572A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2357710Y (en) * | 1999-02-24 | 2000-01-12 | 郑利平 | Disposable lachrymal irrigator |
| CN2560367Y (en) * | 2001-08-30 | 2003-07-16 | 夏勤 | Needle head for dropping medicine suitable for throat and trachea |
| CN201260729Y (en) * | 2008-06-30 | 2009-06-24 | 于青云 | Novel lacrimal passage rinser |
| CN201929962U (en) * | 2010-12-17 | 2011-08-17 | 邱晨 | Blood collection injector |
| CN202776492U (en) * | 2012-04-11 | 2013-03-13 | 上海中医药大学附属曙光医院 | Tail unit curved needle |
| CN103462720A (en) * | 2013-07-24 | 2013-12-25 | 董延庆 | Syringe for veterinarian |
Non-Patent Citations (1)
| Title |
|---|
| 高远等: "脊髓损伤模型的研究进展", 《承德医学院学报》 * |
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Application publication date: 20141029 |
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