CN104996396B - A kind of preservative fluid for perfusing extracorporeal liver - Google Patents
A kind of preservative fluid for perfusing extracorporeal liver Download PDFInfo
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- CN104996396B CN104996396B CN201510444963.5A CN201510444963A CN104996396B CN 104996396 B CN104996396 B CN 104996396B CN 201510444963 A CN201510444963 A CN 201510444963A CN 104996396 B CN104996396 B CN 104996396B
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Abstract
The present invention relates to organ transplantation technique field, specifically disclose the Perfusion preservation liquid of a kind of in vitro liver, including the donor whole proportionally mixed, albumin, antibiotic, insulin, heparin, amino acid, glucose, dexamethasone, sodium chloride, sodium bicarbonate, normal saline, ATP MgCl2, S adenosine egg ammonia；In vitro liver preserves 50 hours in this Perfusion preservation liquid, its hepatocellular damage is few, there is no complication, the demand of organ transplantation can be met, extend the time-to-live of graft, improving graft survival rate in host, Perfusion preservation liquid production cost of the present invention is low, has the using value of reality.
Description
Technical field
The present invention relates to organ transplantation technique field, more particularly, to a kind of isolated organ Perfusion preservation liquid.
Background technology
Organ transplantation is the final treatment means of some diseases, and by organ transplantation, the life of some patients is continued.
In organ transplantation, step the most crucial is the preservation of isolated organ, and this determines the one-tenth of organ transplantation to a great extent
Losing, the organ typically cut from other donors can not be just transplanted in the patient at once, necessarily has one to store transhipment
Process, and at normal temperatures, the cell of isolated organ still provides for anaerobic metabolism, cause intracellular acidosis, mitochondrion production capacity obstacle,
Intracellular calcium overload, cellular swelling, a large amount of free radical generate, and ultimately result in isolated organ and irreversible pathological changes or dead occurs
Die, accordingly, it would be desirable to the isolated organ obtained is carried out suitable Perfusion preservation.
The preservation liquid of the isolated organ generally acknowledged clinically has UW liquid, and its major defect is that its antioxidation is weak, because of UW liquid
In containing containing hetastarch, thus hyperviscosity, bad to isolated organ and lung preservation effect；Application No.
200610124269.3 Chinese patent disclose a kind of novel multiple organs storage protection liquid, this protection liquid apply melatonin,
Vitamin C and the antioxidant system of watermiscible vitamin E composition, it is intended to the oxidation that during reduction organ storage, free radical causes
Damage；The Chinese patent of Application No. 200810036928.X discloses a kind of preservative fluid for perfusing extracorporeal liver, and it is more effective
It is applied to 0～4 DEG C of preservation of in vitro liver, ischemia injury, cellular edema and secondary injury that low temperature causes can be prevented.
It addition, also have a lot of tree-removers can lose owing to the holding time limits in liver transplantation, even if transplant operation
Very successful, but because the damage of hepatic tissue causes preservation process liver mass decline and immunologic rejection causes graft failure, for
Improve the success rate of liver transplantation, it is necessary to develop a kind of Perfusion preservation liquid being directed in vitro liver.
Summary of the invention
Problem to be solved by this invention is to overcome the defect existing for prior art, it is provided that a kind of isolated organ perfusion is protected
Liquid storage.
Purpose of the present invention is achieved by the following technical programs:
A kind of preservative fluid for perfusing extracorporeal liver, every 820mL preserves in liquid containing following component:
Donor whole 500～600mL；
Antibiotic 0.8～2g；
20% albumin 20～50mL；
Insulin 3 00～500u；
Heparin 20000～30000u；
10% amino acid 20～40mL；
Glucose 10～20g；
Dexamethasone 2～10mg；
10% sodium chloride 2～5mL；
5% sodium bicarbonate 20～30mL；
Normal saline 80～120mL；
ATP-MgCl2 50～65g；
S-adenosine egg ammonia 1～3g；
Alprostadil 10～20ug；
Surplus is deionized water, and described amino acid is Amino Acid Compound Injection 18aa.
When carrying out organ transplantation, need the organ cut from donor is carried out temporary transient perfusion and preservation, prior art
In also have organ perfusion preserve liquid, but the Perfusion preservation liquid disclosed in it be only capable of in terms of one solve problem, such as, application number
Be 200610124269.3 Chinese patent disclose a kind of novel multiple organs storage protection liquid, this protection liquid apply take off black
Element, vitamin C and the antioxidant system of watermiscible vitamin E composition, it is intended to during reduction organ storage, free radical causes
Oxidative damage；The Chinese patent of Application No. 200810036928.X discloses a kind of preservative fluid for perfusing extracorporeal liver, and it more has
0～the 4 DEG C of preservation being applied in vitro liver of effect, ischemia injury, cellular edema and the Secondary cases that can prevent low temperature from causing are damaged
Wound；Perfusion preservation liquid of the present invention, for liver transplantation, has explored the new in vitro liver of protection comprehensively, simultaneously can
It is continuously in vitro liver and energy and the Perfusion preservation liquid of metabolism substrate are provided.
Donor whole that the Perfusion preservation liquid of in vitro liver of the present invention includes proportionally mixing, albumin,
Antibiotic, insulin, heparin, amino acid, glucose, dexamethasone, sodium chloride, sodium bicarbonate, normal saline, ATP-
MgCl2, S-adenosine egg ammonia.
Wherein, erythrocyte needed for isolated organ when donor whole can provide external counterpulsation, as oxygen carrier；Simultaneously in whole blood
Blood plasma can maintain the colloid osmotic pressure in infusion liquid；Antibiotic can suppress obtain at organ and enter infusion liquid in filling process
And intraorganic gram negative bacilli, positive cocci and anaerobe.
20% albumin can maintain colloid osmotic pressure in infusion liquid, makes intraor extracellular colloid osmotic pressure keep balance.
Insulin promotion glucose is to intracellular transfer, and promotes protein synthesis；10% amino acid is isolated organ
Protein synthesis provide metabolism substrate；The energy metabolism that 5% glucose is isolated organ provides substrate, is main energy sources；
And 50% glucose is in addition to as energy source, liver cell regeneration can be promoted simultaneously.
Dexamethasone: stabilizing cell membrane and organelle film, alleviates organ inflammation's reaction.
Heparin prevents the blood components in infusion liquid from solidification clot occur, dissolves shape in the tiny blood vessels of perfused organ simultaneously
The thrombosis become；10% sodium chloride maintains the crystalloid osmotic pressure in infusion liquid, it is provided that the sodium ion in infusion liquid；5% sodium bicarbonate can
Neutralize organ in acquisition process and perfusion before produced by anaerobic metabolism a large amount of acidic materials, stablize the PH in infusion liquid
Value.
Normal saline: increase infusion liquid capacity, as the primary solvent of infusion liquid, can provide stable crystalline percolation simultaneously
Pressure, makes infusion liquid be in isotonic state；ATP-MgCl2: energy is directly provided to cell, recovers the energy storage that organ cell is certain
Standby, make the organ-tissue ATP content of hot ischemia increase；Stabilizing cell membrane and organelle film, reduce permeability of cell membrane, alleviates thin
Born of the same parents' edema；Improve the microcirculation after resurgent and regional perfusion amount, thus alleviate hypoxia and the low-energy state of hepatic tissue；S-adenosine egg
Ammonia: can improve hepatocyte DNA methylation level, promotes the synthesis of DNA, inducing hepatocyte mitosis in early days, promotees its regeneration, with
Time be conducive to the generation of intracellular ATP；Blood vessel in the expansible organ of Alprostadil, improves perfusion blood flow；Maintain crystalloid osmotic pressure
And colloid osmotic pressure.
Preferably, above-mentioned Perfusion preservation liquid also includes 5～10mL 10% calcium gluconate；Calcium gluconate can be safeguarded carefully
After birth and the normal permeability of blood capillary, so that liver cell is preferably immersed in Perfusion preservation liquid.
Applicant, during compounding above-mentioned Perfusion preservation liquid, finds that the Alprostadil proportioning with calcium gluconate is to filling
The preservation effect impact of fluid injection is notable, it is preferable that only the amount ratio of Alprostadil and 10% calcium gluconate is 10～16ug:6
～during 10mL, the in vitro liver holding time in described Perfusion preservation liquid is long, and the harmful effect to liver is minimum.
Preferably, in above-mentioned preservation liquid, every 820mL preserves in liquid containing following component:
Donor whole 500～520mL；
Antibiotic 0.8～1.5g；
20% albumin 25～35mL；
Insulin 400～420u；
Heparin 25000～28000u；
10% amino acid 25～30mL；
Glucose 10～18g；
Dexamethasone 4～10mg；
10% sodium chloride 2～5mL；
5% sodium bicarbonate 25～30mL；
Normal saline 80～100mL；
ATP-MgCl2 60～65g；
S-adenosine egg ammonia 1～3g；
Alprostadil 10～16ug；
10% calcium gluconate 6～10mL
Surplus is deionized water.
Preferably, described antibiotic is selected from one or both in metronidazole, cefazolin.
There is provided any one liver perfusion above-mentioned to preserve liquid and preserve the application in preparation in preparation organ transplantation.
Compared with prior art, the method have the advantages that
The invention provides the Perfusion preservation liquid of a kind of in vitro liver, including the donor whole proportionally mixed,
Albumin, antibiotic, insulin, heparin, amino acid, glucose, dexamethasone, sodium chloride, sodium bicarbonate, physiology salt
Water, ATP-MgCl2, S-adenosine egg ammonia；In vitro liver preserves 50h in this Perfusion preservation liquid, and hepatocyte only has slight swelling,
Remaining is normal, does not has complication, can meet the demand of organ transplantation, extends the time-to-live of graft, improves graft and exists
Survival rate in host, Perfusion preservation liquid production cost of the present invention is low, has the using value of reality.
Accompanying drawing explanation
Fig. 1 is each group of HE coloration result.；A is UW perfusion group hepatic tissue；B is UW perfusion group bile duct tissue；C is for filling
Fluid injection perfusion group hepatic tissue；D is infusion liquid perfusion group bile duct tissue.
Detailed description of the invention
Below by Figure of description and specific embodiment specific descriptions further to the present invention.Following used experiment
If method is without specified otherwise, the method being the existing routine of the art, the dispensing used or material, as without special theory
Bright, it is by the available dispensing of commercial sources or material；The present invention should not necessarily be limited to scope of embodiments.
Embodiment 1
The compound method of the present embodiment infusion liquid: under aseptic condition, takes portions of de-ionized water, will be except donor whole and white egg
Remaining material outside Bai is firstly added in deionized water, adds albumin and donor whole, be more finally settled to 820 after mixing
mL。
Every 820mL infusion liquid includes each component in table 1:
Embodiment 2
The compound method of the present embodiment infusion liquid: under aseptic condition, takes portions of de-ionized water, will be except donor whole and white egg
Remaining material outside Bai is firstly added in deionized water, adds albumin and donor whole, be more finally settled to 820 after mixing
mL。
Every 820mL infusion liquid includes each component in table 2:
Embodiment 3
The compound method of the present embodiment infusion liquid: under aseptic condition, takes portions of de-ionized water, will be except donor whole and white egg
Remaining material outside Bai is firstly added in deionized water, adds albumin and donor whole, be more finally settled to after mixing
820mL。
Every 820mL infusion liquid includes each component in table 3:
Comparative example 1
Experimental technique is with embodiment 1, without S-adenosine egg ammonia and Portugal in infusion liquid described in unique except for the difference that this comparative example
Grape Calciofon, the cumulative volume of infusion liquid is 820mL.
Comparative example 2
Experimental technique is with embodiment 1, without S-adenosine egg ammonia, perfusion in infusion liquid described in unique except for the difference that this comparative example
The cumulative volume of liquid is 820mL.
Comparative example 3
Experimental technique is with embodiment 1, without calcium gluconate, perfusion in infusion liquid described in unique except for the difference that this comparative example
The cumulative volume of liquid is 820mL.
Comparative example 4
Experimental technique with embodiment 1～3, unique unlike Alprostadil and 10% Portugal in infusion liquid described in this comparative example
The amount ratio of grape Calciofon is 20ug:10mL.
Comparative example 5
Experimental technique with embodiment 1～3, unique unlike Alprostadil and 10% Portugal in infusion liquid described in this comparative example
The amount ratio of grape Calciofon is 10ug:20mL.
Comparative example 6
Experimental technique with embodiment 1, unique unlike, the amino acid used by this comparative example is Patent No.
200810036928.X Chinese patent described in the aminoacid used in preservative fluid for perfusing extracorporeal liver.
Comparative example 7
Experimental technique with embodiment 1, unique unlike, this comparative example artificial blood replaces donor whole.
Comparative example 8
Experimental technique with embodiment 1, unique unlike, this comparative example erythrocyte replaces donor whole.
Embodiment 4
Infusion liquid described in embodiment 1～3 and comparative example 1～8 is used for the Perfusion preservation of liver, observes the preservation shape of liver
State, its result such as table 4.
Embodiment 5
1, laboratory animal
Male Tibet Mini-Pigs 10, regular grade, body weight 20～25kg, monthly age 4～June, it is purchased from Nanfang Medical Univ real
Test animal center.Receptacle room temperature whole day is controlled in 20～25 DEG C, and fed standard chow is freely drunk water.Animal is all in accordance with country
Regulation carries out hommization raising, it is ensured that the relevant welfare of laboratory animal, is used for testing after raising 3 to 7 days.
2, experiment packet and model are set up:
All animals are randomly divided into following 2 groups before experiment: preserve liquid (selecting to preserve described in embodiment 1 liquid) perfusion
Group, UW perfusion group, often group 5.Concrete model method for building up is as follows:
Laboratory animal anesthesia posterior vein bolus 10% potassium chloride injection 40ml.Treat that electrocardio-activity disappears and tranquillization 5 points
Zhong Hou, we are judged to heart death.And carried out liver acquisition again after full 90 minutes of death time.Row room temperature machine subsequently
Tool irrigates 4 hours also whole blood perfusion resurgent 1 hours.Whole blood row Liver function grade after leaving and taking Pathologic specimen subsequently and gathering resurgent,
Result such as table 5.
HE coloration result such as Fig. 1, shows similar with normal bile duct tissues, epithelial duct under infusion liquid group bile duct tissue light microscopic
Cell more complete (Fig. 1 D).UW group is after the whole blood resurgent of 1 hour, and structural deterioration occurs in biliary tract, and pathological manifestations is: common bile duct gallbladder
Destroying occurs in road epithelial integrity, and biliary tract epithelial cell major part is peeled off and disappeared, and on biliary tract, subcutaneous connective tissue edema is obvious, complete
Naked dew (Figure 1B).Infusion liquid group pathological manifestations under light microscopic is: popularity edema occurs in hepatocyte, and sinus hepaticus gap is broadening,
Hyperemia, hepatocyte existence is dispersed in necrotic zone (Figure 1A).UW liquid group is identical with perfusion group pathological change type, but performance is the tightest
Weight (Fig. 1 C).
Claims (3)
1. a preservative fluid for perfusing extracorporeal liver, it is characterised in that every 820mL preserves in liquid containing following component:
Donor whole 500～520mL；
Antibiotic 0.8～1.5g；
20% albumin 25～35mL；
Insulin 400～420u；
Heparin 25000～28000u；
10% amino acid 25～30mL；
Glucose 10～18g；
Dexamethasone 4～10mg；
10% sodium chloride 2～5mL；
5% sodium bicarbonate 25～30mL；
Normal saline 80～100mL；
ATP-MgCl2 60～65g；
S-adenosine egg ammonia 1～3g；
Alprostadil 10～16ug；
10% calcium gluconate 6～10mL
Surplus is deionized water.
Liver perfusion the most according to claim 1 preserves liquid, it is characterised in that described antibiotic is selected from metronidazole, head
One or both in spore azoles woods.
3. liver perfusion described in any one of claim 1 to 2 preserves liquid and preserves the application in preparation in preparation organ transplantation.
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Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105532646B (en) * | 2016-01-27 | 2017-10-13 | 武汉仝干医疗科技股份有限公司 | Liver cell preserves liquid and its preparation method and application |
| CN105918308A (en) * | 2016-04-28 | 2016-09-07 | 上海市胸科医院 | Lung perfusion fluid for perfusing transplant donor lung and preparation method for lung perfusion fluid |
| CN109511650B (en) * | 2018-11-21 | 2021-06-15 | 嘉兴莱普晟医疗科技有限公司 | Normal-temperature mechanical perfusate capable of enlarging liver supply source |
| CN109511649B (en) * | 2018-11-21 | 2021-06-25 | 嘉兴莱普晟医疗科技有限公司 | Normal-temperature mechanical perfusion system capable of expanding liver supply source |
| CN112425602A (en) * | 2019-08-26 | 2021-03-02 | 无锡市人民医院 | Isolated lung mechanical perfusion liquid and preparation method and application thereof |
| CN112273373B (en) * | 2020-10-29 | 2022-06-07 | 无锡市人民医院 | Isolated lung mechanical perfusion liquid and preparation method and application thereof |
| CN113287602B (en) * | 2021-05-28 | 2022-09-30 | 吉林大学第一医院 | Normal-temperature mechanical perfusate and liver in-vitro preservation method |
| CN113498777B (en) * | 2021-07-26 | 2022-12-30 | 公卫复医(上海)生物科技有限公司 | Emulsion type organ preservation solution and preparation method thereof |
| CN115715540B (en) * | 2022-11-18 | 2024-04-02 | 中山大学附属第一医院 | Mechanical perfusate and preparation method and application thereof |
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