CN110384569A - A kind of preparation method of recoverability obstruction of bile duct rat model - Google Patents
A kind of preparation method of recoverability obstruction of bile duct rat model Download PDFInfo
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- CN110384569A CN110384569A CN201910671946.3A CN201910671946A CN110384569A CN 110384569 A CN110384569 A CN 110384569A CN 201910671946 A CN201910671946 A CN 201910671946A CN 110384569 A CN110384569 A CN 110384569A
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Abstract
The application belongs to experimental animal field, and in particular to a kind of Vein Tube needle for the artificial biliary tract building of rat and a kind of preparation method of recoverability obstruction of bile duct rat model.Vein Tube needle its end of the present invention for the artificial biliary tract building of rat is in T-shape, can be used for the building of the artificial biliary tract of rat.The preparation method of recoverability obstruction of bile duct rat model of the present invention constructs the artificial biliary tract of rat in T-shape Vein Tube needle using end；The Vein Tube needle is ligatured to form obstruction of bile duct using suture, ligation is released and then releases obstruction of bile duct, recoverability obstruction of bile duct rat model is made.The present invention can also carry out required other operations to hepatobiliary system in the artificial biliary tract of rat that the Vein Tube needle of T-shape is established using the end.Preparation method of the present invention is easy to operate, does not destroy pancreas in rat, ductus pancreaticus and left and right common hepatic duct structure；It is not easily to fall off to cause leak or intestinal fistula；Need consumptive material few, it is at low cost.
Description
Technical field
The invention belongs to experimental animal fields, and in particular to a kind of preparation method of recoverability obstruction of bile duct rat model.
Background technique
Bile duct, biliary pipeline.Digestion food Shi ﹐ is stored in the bile in gall-bladder and flows into duodenum by it.
Obstruction of bile duct (obstruction of biliary tract) refer to any one section of bile outlet duct because of calculus of bile duct, bile duct chamber
The diseases such as compressing are infiltrated outside interior lesion, tube wall self-disease, tube wall, cause bile drainage difficulty or even completely plugged bile duct
Mechanical obstruction.Typical clinical manifestation is jaundice, and when secondary infection of biliary tract such as can generate heat at the symptoms with abdominal pain and chilly.Biliary tract
Obstruction is a kind of disease relatively conventional in Surgical Acute Venter Diseases, most secondary to calculus of bile duct, but stenosis of bile duct and tumor of bile duct
Etc. diseases also can this secondary disease.Above-mentioned disease can cause bile duct obstruction, cholestasis and secondary bacterial infection.Pathogenic bacteria master
Enteron aisle is come from, is driven in the wrong direction through the ampulla of vater or through Cholangio- jejunostomy mouth into biliary tract.Pathogenic bacteria also can enter biliary tract through hematogenous infection.
In order to preferably carry out bile duct disease research, need to establish recoverability obstruction of bile duct animal model.
The animal model (Animal Model of Human Diseases) of human diseases is biomedical sciences research
Middle the established zoopery object and material with human diseases simulation sex expression.It the use of animal model is that modern biotechnology is cured
Particularly important an experimental method and means in research are learned, helps to be more convenient, more effectively recognize the hair of human diseases
The raw, rule of development and research control measure.Experimental study needs ideal animal model, since larger animal obtains difficult, feeding
The reasons such as inconvenience are supported, its application in liver transfer operation experimental study is limited.In recent years, rat has become liver transfer operation experimental study
Preferred animal this is because rat has following advantages: 1. the progress of microsurgical technique is so that Establishment of Rat Liver Transplantation Model
Success rate improve；2. the acquisition of homogenic source rat is easier to；3. rat is easy to raise；4. the natural resistance bacterium sense of rat
Dye ability is stronger；5. the expense of raising is lower etc..
However rat is the experimental animal of no gall-bladder, Common Bile Duct Diameter is typically only 0.5-0.8mm；In addition, choledochus
It is wrapped completely by the pancreas of sheet with left and right common hepatic duct, and ductus pancreaticus number and conduct are different because of individual, it partially can be at different level
It being open into hepatic duct, therefore when establishing recoverability obstruction of bile duct rat model, existing operation method difficulty is big, process is complicated,
Such as the bile duct that excessively dissociates, it is easy to damage ductus pancreaticus and pancreas, destroys the normal exocrine function of pancreas.Such as " improvement recoverability obstruction
Property jaundice Establishment of Rat Model and its application value " (Chen Fangfang etc., Chinese liver surgery e-magazine 2 months 2019
The 1st phase of volume 8) " free choledochus 5mm " is needed described in the model established at it in a text, and need " by choledochus, defeated
Liquid pipe, the folded junction of gasket three's row dual three are pricked ", in conjunction with above-mentioned big cowardly pancreas anatomical structure feature, this method is easy to damage pancreas,
And operating process is complicated." foundation and evaluation of recoverability Jaundice of Rats model " (Fang Hui etc., world Chinese digest magazine
On September 8th, 2015；23 (25): 4015-4023) it establishes " bile duct ligation support tube pulls out method " described in need " trip
At bile duct to hepatic portal ", it needs using " PICC conduit " and " the multiple triumphant stoma of jejunum paracentesis trocar of that ", and need " by PICC
Both conduit and choledochus row three folds junction bundle ", this method operating process is complicated, and easy damaged pancreas, in addition, the equipment number used
Amount is more and costly, is unfavorable for promoting." microsurgical technique establishes drainage model in stable experimental rat obstructive jaundice " (Liu Han
Raw etc., contemporary Chinese general surgery is in progress in November, 2017 o. 11th 845-850 of volume 20) described in the model established at it
" needing to carry out cystic duct cannula twice to rat ", and " need to be intubated free-end after first time is intubated and be fixed on stomach wall
On ", in actual operation, complicated operation for this, and intubation free-end is fixed on stomach wall, intubation is easily caused to fall off, thus
Cause serious leak." a kind of foundation of improved obstructive jaundice rats model " (Chen Dong etc. digests tumour magazine electronic
Version, the 12nd phase of volume 1 in December, 2009) it needs " by outside drainage tube free-end lead body, to be fixed on flesh in the model that it is established
In meat and skin ", and " external drainage " is carried out to rat after releasing obstruction, complicated operation for this method, and intubation is swum
It easily causes intubation set terminal to fall off from the way that integumentary musculature is fixed at end, causes leak, in addition, external drainage and not meeting normal
Physiological status.And a kind of Chinese patent (CN105055040A) of the construction method of entitled pancreatic duct obstruction in rats model
Described in " with closed remaining needle by duodenum insertion pancreatic duct realize obstruction, pull out realization release obstruction ", in conjunction with above
The rat biliary anatomy, this method is more difficult in actual operation, and has bile to overflow and cause rat intestine from intubation periphery
The possibility of obstruction.
Summary of the invention
In view of this, it is an object of the present invention to for it is existing in the prior art it is complicated for operation, be easy to cause pancreas,
The problem of organ damages such as ductus pancreaticus, provides a kind of preparation method of recoverability obstruction of bile duct rat model, to simplify operating process,
Avoid the organ damages such as pancreas, ductus pancreaticus.
To achieve the purpose of the present invention, the present invention adopts the following technical scheme:
Present invention firstly provides a kind of Vein Tube needles for the artificial biliary tract building of rat, and end is in T-shape.
The present invention utilizes Vein Tube needle, is transformed to it, makes its end in T-shape.It can be used for the artificial biliary tract of rat
Building.
Preferably, described its specification of Vein Tube needle is 22G*1.00in.
The present invention also provides a kind of preparation methods of recoverability obstruction of bile duct rat model, using above-mentioned Vein Tube
Needle constructs the artificial biliary tract of rat；The Vein Tube needle is ligatured to form obstruction of bile duct using suture, ligation is released and then releases
Recoverability obstruction of bile duct rat model is made in obstruction of bile duct.
Preferably, the suture for ligaturing Vein Tube needle described in the preparation method is 2-0 suture.
Preferably, the method for constructing the artificial biliary tract of rat described in the preparation method specifically: rat is through anaesthetizing
Afterwards, take dorsal position to fix, slough abdomen chaeta, disinfection, taken by xiphoid-process ing the abundant exposed Rats duodenum in 3cm abdomen center,
Pancreas, bile duct and left and right common hepatic duct, away from choledochus is ligatured at the common hepatic duct meet 1.5cm of left and right, vein is inserted into proximal end at ligation
Pipe needle is set, is fixed after placed-depth 1cm, T-shape then is made in Vein Tube needle end, by T-type end merging bile duct opening
In neighbouring duodenum, intestinal tube is sutured.
Wherein, the method for the anesthesia is to give 2% sodium phenobarbital according to the abdominal cavity 0.3ml/100g according to rat body weight
Solution.
Preferably, ligation choledochus described in the method for the building artificial biliary tract of rat uses 7-0 suture ligature.
Preferably, the Vein Tube needle 7-0 suture at ligation described in the method for the building artificial biliary tract of rat
It is fixed with biological glue.
Preferably, the every valve of T-shape of Vein Tube needle end described in the method for the building artificial biliary tract of rat is long
Spend 0.2cm.After the i.e. described Vein Tube pin intubation to rat choledochus, extracts needle core out, T-shape is made in end with eye scissors,
Cutting off length is 0.2cm.
Preferably, suture intestinal tube described in the method for the building artificial biliary tract of rat uses 8-0 suture.
As shown from the above technical solution, the present invention provides a kind of Vein Tube needle for rat artificial biliary tract building with
And a kind of preparation method of recoverability obstruction of bile duct rat model.Vein of the present invention for the artificial biliary tract building of rat is set
Its end of pipe needle is in T-shape, can be used for the building of the artificial biliary tract of rat.Recoverability obstruction of bile duct rat model of the present invention
Preparation method constructs the artificial biliary tract of rat in T-shape Vein Tube needle using end；Using suture to the Vein Tube needle
Ligation forms obstruction of bile duct, releases ligation and then releases obstruction of bile duct, recoverability obstruction of bile duct rat model is made.It is of the present invention
The preparation method of recoverability obstruction of bile duct rat model by the Vein Tube needle being ligatured and being released the operation of ligation,
The recoverability obstruction of bile duct to experimental rat may be implemented, additionally can use the Vein Tube needle that the end is in T-shape
The artificial biliary tract of the rat established carries out required other operations to hepatobiliary system, as realized rat bile duct and any intestinal segment
Cholangio- jejunostomy and to experiment process such as the administrations, drainage, obstruction of rat hepatobiliary system.Recoverability bile duct stalk of the present invention
The preparation method for hindering rat model is easy to operate, does not destroy pancreas in rat, ductus pancreaticus and left and right common hepatic duct structure；End is in T-shape
Vein Tube needle is fixed, not easily to fall off to cause leak or intestinal fistula；Need consumptive material few, it is at low cost.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technical description to be briefly described.
Fig. 1 is schematic diagram of the end in the Vein Tube needle of T-shape, and A is Vein Tube needle, and B is extraction needle core, uses ophthalmology
Cut the Vein Tube needle that end is made to T-shape；
Fig. 2 is recoverability obstruction of bile duct rat model building process schematic diagram；
Fig. 3 is each step picture of recoverability obstruction of bile duct rat model building process；
Fig. 4 is not block group and obstruction 24 hours groups, first day after surgery living imaging figure；
Fig. 5 is to block 24 hours group first day after surgery, second day and the 7th day living imaging figures of difference.
Specific embodiment
The invention discloses a kind of preparation methods of recoverability obstruction of bile duct rat model.Those skilled in the art can borrow
Reflect present disclosure, is suitably modified realization of process parameters.In particular, it should be pointed out that all similar substitutions and modifications are to this field
It is it will be apparent that they are considered as being included in the present invention for technical staff.Method and product of the invention has passed through
Preferred embodiment is described, related personnel obviously can not depart from the content of present invention, in spirit and scope to described herein
Method be modified or appropriate changes and combinations, carry out implementation and application the technology of the present invention.
For a further understanding of the present invention, below in conjunction with the embodiment of the present invention, to the technical side in the embodiment of the present invention
Case is clearly and completely described, it is clear that and described embodiments are only a part of the embodiments of the present invention, rather than all
Embodiment.Based on the embodiments of the present invention, those of ordinary skill in the art institute without making creative work
The every other embodiment obtained, shall fall within the protection scope of the present invention.
Unless otherwise specified, reagent involved in the embodiment of the present invention is commercial product, can pass through business canal
Road purchase obtains.
Embodiment 1,
1. experimental animal and equipment:
(1) experimental animal: purebred Sprague-Dawley rat, is male, and SPF grades, 6-7 week old, weight 200-
250g.Animal shakes down 7 days before testing, and every 2 cages are respectively placed in cage the environment raised, kept constant, temperature
18~20 DEG C of degree, humidity 60%, rat arbitrarily drink water and feed daily.At one night of pre-operative anxiety, it can't help water；
(2) experiment reagent: sodium phenobarbital (Sigma company), D- fluorescein sylvite (Promega company), luciferase
Reporter plasmid (pMIR-REPORT Luciferase plasmid DNA, hereinafter referred to as " LUC plasmid ", by Nanjing University
Medical biotechnology National Key Laboratory doctor Zhang Yifan grants)；
(3) crucial equipment: Vein Tube needle (22G*1.00in, BD company) is tested；
(4) operating equipment: eye scissors, ophthalmic tweezers, ophthalmology needle holder, skin pincers, Self-made hook, biological glue, suture needle, suture
(2-0,7-0,8-0), insulin needle (29G*1/2in, BD company)；
(5) living body visible light 3-D imaging system (IVIS Spectrum).
2. making by oneself T-shape tube (Fig. 1, hereinafter referred to as " T pipe ")
Vein Tube needle (22G*1.00in, U.S. company BD) is transformed: after intubation to rat bile duct, extracting needle out
With eye scissors T-shape is made in end by core.
3. construction method:
(1) anaesthetize: anesthesia: according to rat body weight, dosage 0.3ml/100g gives 2% sodium phenobarbital
Solution intraperitoneal injection.
(2) building process (Fig. 2 and Fig. 3):
After anaesthetizing successfully, rat takes supine position on rat operation plate, fixing teeth and four limbs.It is taken off greatly with depilatory cream
Then mouse abdomen chaeta is sterilized liquid disinfectant abdomen three times with povidone-iodine.By xiphoid-process, 3cm abdomen median incision is taken, coagulation is stopped
Blood.Sufficiently (do not dissociate gallbladder for exposed Rats duodenum, pancreas, bile duct and left and right common hepatic duct with the help of skin pincers and Self-made hook
Pipe, ductus pancreaticus and left and right common hepatic duct) (Fig. 2 and Fig. 3 A).At away from left and right common hepatic duct meet 1.5cm, with 7-0 suture ligature choledochus,
Visible proximal bifiary ducts fill (Fig. 2 and Fig. 3 B) after ligation.Under a dissecting microscope, Vein Tube is inserted into proximal end at ligation, sets
Enter depth 1cm, visible end has faint yellow limpid bile to draw after being successfully placed in.It is fixed immediately with 7-0 suture and biological glue,
Then T-shape, every valve length 0.2cm (Fig. 2 and Fig. 3 C) is made in end with eye scissors.Bile duct is open neighbouring with eye scissors
Duodenum intestinal tube does the notch of 1-2mm, then follows this notch and T-type end is placed in duodenum, sutures intestines with 8-0 suture
It manages (Fig. 2 and Fig. 3 D).
Thus the artificial biliary tract of rat is established.On this basis, ligation is carried out to T pipe using 2-0 suture and forms bile duct
Obstruction；And can be needed to release the ligature on T pipe in corresponding time row second operation according to experiment, to release obstruction of bile duct
(Fig. 2 and Fig. 3 D).
Embodiment 2,
For the feasibility for verifying this model, LUC plasmid is injected using through artificial biliary tract antidromicity in this experiment, according to it
The case where fluorescence detection result judgement obstruction and releasing obstruction.
After success constructs artificial biliary tract, insulin needle is entered from the duodenum intestinal wall of T tube opening opposite side, insertion T pipe
It is interior.Then by LUC plasmid (50 μ g LUC plasmids/1ml distilled water), fast reverse injects rat hepatic duct in 5s.Injection finishes
Afterwards, insulin needle is exited.Duodenum intestinal wall is sutured with 8-0 suture when necessary.
After exiting insulin needle, T pipe is ligatured immediately and forms obstruction of bile duct.It is closed after confirming intraperitoneal no bleeding with 2-0 suture
Abdomen.Free water (5% syrup and distilled water) is given in postoperative fasting 24 hours.Every rat is injected intraperitoneally 20,000 2 times a day simultaneously
IU penicillin is anti-infective, and continuous 5 days.24 hours postoperative (obstruction of bile duct 24 hours) row first time living imaging (Fig. 4).After imaging
In anaesthetizing lower laparotomy exploration, the ligature on decannulation is solved, closes abdomen after confirmation bile excretion is unobstructed.Then respectively at the postoperative 2nd
Row living imaging (Fig. 5) again of it and postoperative 7th day.
24 hours rats of obstruction of bile duct are 4 total in experiment.According to experiment needs, while to other 4 rat pedestrians
The building of work biliary tract, does not give obstruction, as experiment reference.
By Fig. 4 result as it can be seen that first post-operative day testing result is shown: not blocking group: photon amount (3.94 ± 0.70) * 106；Stalk
Hinder 24 hours groups: photon amount (1.56 ± 0.42) * 108(p=0.036).Show that the method for the invention can effectively realize rat
Obstruction of bile duct.
By Fig. 5 result as it can be seen that showing the obstruction continuous row living imaging result of 24 hours groups: first post-operative day POD1: photon
Measure (1.56 ± 0.42) * 108, second day after operation POD2: photon amount (3.42 ± 0.54) * 106, postoperative 7th day POD7: photon amount
(3.72±0.62)*106；1 versus POD 2, p=0.002 of POD；1 versus POD 7, p=0.002 of POD；POD
2 versus POD 7, p=0.993.Show that the method for the invention after effectively realizing rat obstruction of bile duct, can be released effectively
The recoverability obstruction of rat bile duct is realized in obstruction.
Claims (9)
1. a kind of Vein Tube needle for the artificial biliary tract building of rat, end is in T-shape.
2. Vein Tube needle according to claim 1, specification 22G*1.00in.
3. a kind of preparation method of recoverability obstruction of bile duct rat model, using Vein Tube needle structure of any of claims 1 or 2
Build the artificial biliary tract of rat；The Vein Tube needle is ligatured to form obstruction of bile duct using suture, ligation is released and then releases bile duct
Recoverability obstruction of bile duct rat model is made in obstruction.
4. the suture of preparation method according to claim 3, the ligation Vein Tube needle is 2-0 suture.
5. preparation method according to claim 3, the method for the building artificial biliary tract of rat specifically: rat is through anaesthetizing
Afterwards, take dorsal position to fix, slough abdomen chaeta, disinfection, taken by xiphoid-process ing the abundant exposed Rats duodenum in 3cm abdomen center,
Pancreas, bile duct and left and right common hepatic duct, away from choledochus is ligatured at the common hepatic duct meet 1.5cm of left and right, vein is inserted into proximal end at ligation
Pipe needle is set, is fixed after placed-depth 1cm, T-shape then is made in Vein Tube needle end, by T-type end merging bile duct opening
In neighbouring duodenum, intestinal tube is sutured.
6. preparation method according to claim 5, the method for the anesthesia is according to rat body weight according to 0.3ml/100g
Give 2% phenobarbital sodium solution in abdominal cavity.
7. preparation method according to claim 5, the ligation choledochus uses 7-0 suture ligature；At the ligation
Vein Tube needle is fixed with 7-0 suture and biological glue.
8. preparation method according to claim 5, the every valve length 0.2cm of the T-shape of Vein Tube needle end.
9. preparation method according to claim 5, the suture intestinal tube uses 8-0 suture.
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| CN1466438A (en) * | 2000-09-29 | 2004-01-07 | ��ʿ�ٿƼ�����˾ | Method for performing endoluminal fundoplication and apparatus for use in the method |
| US20080015416A1 (en) * | 2006-07-14 | 2008-01-17 | Wilson-Cook Medical, Inc | Papilla spreader |
| US20100137891A1 (en) * | 2007-06-04 | 2010-06-03 | Svip 8Llc | Tissue anchorable devices |
| CN102056551A (en) * | 2008-05-01 | 2011-05-11 | 伊西康内外科公司 | Method and apparatus for the formation of tissue folds |
| CN201356684Y (en) * | 2009-01-10 | 2009-12-09 | 田明国 | Soft bile duct support |
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Application publication date: 20191029 |