CN115443947B - Preparation method of hypertension animal model - Google Patents
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- CN115443947B CN115443947B CN202211244294.3A CN202211244294A CN115443947B CN 115443947 B CN115443947 B CN 115443947B CN 202211244294 A CN202211244294 A CN 202211244294A CN 115443947 B CN115443947 B CN 115443947B
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- 238000011616 hypertension animal model Methods 0.000 title claims abstract description 8
- 238000002360 preparation method Methods 0.000 title claims abstract description 5
- 206010020772 Hypertension Diseases 0.000 claims abstract description 34
- 230000036772 blood pressure Effects 0.000 claims abstract description 17
- 210000000028 corpus adiposum pararenale Anatomy 0.000 claims abstract description 15
- 239000000126 substance Substances 0.000 claims abstract description 14
- 241001465754 Metazoa Species 0.000 claims abstract description 12
- YKPUWZUDDOIDPM-SOFGYWHQSA-N capsaicin Chemical compound COC1=CC(CNC(=O)CCCC\C=C\C(C)C)=CC=C1O YKPUWZUDDOIDPM-SOFGYWHQSA-N 0.000 claims description 16
- 238000000034 method Methods 0.000 claims description 11
- ZKHQWZAMYRWXGA-UHFFFAOYSA-N Adenosine triphosphate Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)C(O)C1O ZKHQWZAMYRWXGA-UHFFFAOYSA-N 0.000 claims description 8
- 102000004889 Interleukin-6 Human genes 0.000 claims description 8
- 108090001005 Interleukin-6 Proteins 0.000 claims description 8
- 238000010171 animal model Methods 0.000 claims description 8
- 229960002504 capsaicin Drugs 0.000 claims description 8
- 235000017663 capsaicin Nutrition 0.000 claims description 8
- 239000002671 adjuvant Substances 0.000 claims description 6
- ZKHQWZAMYRWXGA-KQYNXXCUSA-J ATP(4-) Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP([O-])(=O)OP([O-])(=O)OP([O-])([O-])=O)[C@@H](O)[C@H]1O ZKHQWZAMYRWXGA-KQYNXXCUSA-J 0.000 claims description 4
- 229940100601 interleukin-6 Drugs 0.000 claims description 4
- 230000002146 bilateral effect Effects 0.000 claims description 3
- 238000002347 injection Methods 0.000 claims description 3
- 239000007924 injection Substances 0.000 claims description 3
- 238000005086 pumping Methods 0.000 claims description 3
- 230000003442 weekly effect Effects 0.000 claims description 3
- 230000000638 stimulation Effects 0.000 abstract description 5
- 230000002269 spontaneous effect Effects 0.000 abstract description 4
- 230000004936 stimulating effect Effects 0.000 abstract description 3
- 230000007547 defect Effects 0.000 abstract description 2
- 230000035772 mutation Effects 0.000 abstract description 2
- 230000008506 pathogenesis Effects 0.000 abstract description 2
- 238000011552 rat model Methods 0.000 abstract description 2
- 230000000630 rising effect Effects 0.000 abstract description 2
- 229940043263 traditional drug Drugs 0.000 abstract description 2
- 206010065918 Prehypertension Diseases 0.000 abstract 1
- 241000700159 Rattus Species 0.000 description 10
- 238000000520 microinjection Methods 0.000 description 6
- 238000000465 moulding Methods 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 5
- 238000001802 infusion Methods 0.000 description 5
- 239000011780 sodium chloride Substances 0.000 description 5
- 238000009530 blood pressure measurement Methods 0.000 description 4
- 235000005911 diet Nutrition 0.000 description 3
- 230000037213 diet Effects 0.000 description 3
- 229910052751 metal Inorganic materials 0.000 description 3
- 239000002184 metal Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 208000007530 Essential hypertension Diseases 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 235000009200 high fat diet Nutrition 0.000 description 2
- 230000001631 hypertensive effect Effects 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 102000015427 Angiotensins Human genes 0.000 description 1
- 108010064733 Angiotensins Proteins 0.000 description 1
- 241000282414 Homo sapiens Species 0.000 description 1
- 208000008589 Obesity Diseases 0.000 description 1
- 201000004239 Secondary hypertension Diseases 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000004872 arterial blood pressure Effects 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000003246 corticosteroid Substances 0.000 description 1
- 229960001334 corticosteroids Drugs 0.000 description 1
- 230000035487 diastolic blood pressure Effects 0.000 description 1
- 235000021196 dietary intervention Nutrition 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 230000000214 effect on organisms Effects 0.000 description 1
- 239000003862 glucocorticoid Substances 0.000 description 1
- 235000021070 high sugar diet Nutrition 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 239000002395 mineralocorticoid Substances 0.000 description 1
- 235000020824 obesity Nutrition 0.000 description 1
- 230000003362 replicative effect Effects 0.000 description 1
- 235000015598 salt intake Nutrition 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 230000035488 systolic blood pressure Effects 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/02—Breeding vertebrates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/10—Mammal
- A01K2227/105—Murine
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/03—Animal model, e.g. for test or diseases
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/03—Animal model, e.g. for test or diseases
- A01K2267/035—Animal model for multifactorial diseases
- A01K2267/0375—Animal model for cardiovascular diseases
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a preparation method of a hypertension animal model, which is to inject chemical stimulating substances into perirenal fat of animals. The invention only adopts a single intrarenal fat administration stimulation substance, and a stable hypertension animal model is obtained after 4 weeks, overcomes the defects of low modeling rate, unstable model and the like of the traditional drug-induced model, has the characteristics of 100 percent modeling rate, adjustable blood pressure rising amplitude and the like, and can obtain a hypertension model with obviously raised blood pressure and a pre-hypertension model with slightly raised blood pressure by controlling the dosage of the stimulation substance. Compared with the existing spontaneous hypertension rat model, the invention avoids early-onset and inherent hypertension caused by genetic mutation and the like, is more similar to the clinical hypertension pathogenesis, and the grading type hypertension model of the invention cannot be realized by the spontaneous hypertension model.
Description
Technical Field
The invention relates to a preparation method of a hypertension animal model, and belongs to the field of disease animal models.
Background
The cause of essential hypertension is quite complex, and it is only known that various common risk factors including high sodium salt intake, obesity, metabolic disorders and the like can promote the occurrence of hypertension. However, the single or mixed action of these risk factors can also induce stable hypertension in only a subset of animals and humans. Taking animal models as an example, high salt diets and high fat and high sugar diets for a long period of time (usually 10-16 weeks) induce rats with a incidence of hypertension of less than 50%, and after stopping the diets, the animals' hypertension mostly can be significantly reduced or restored to normal levels. It is clear that such a purely dietary intervention method is not effective in replicating hypertension occurring in humans. The hypertension model induced by the booster hormone substances such as glucocorticoid, mineralocorticoid, angiotensin and the like can only simulate the secondary hypertension of human beings, and cannot respond to the formation process of the primary hypertension and adverse effects on organisms.
Disclosure of Invention
The invention aims to provide a method for stably preparing a hypertension animal model.
The technical scheme adopted by the invention is as follows: a method for preparing animal model of hypertension comprises injecting chemical stimulating substance into animal perirenal fat.
Preferably, the chemical stimulating substance is capsaicin, complete Freund's adjuvant, adenosine triphosphate, interleukin 6. The administration may be continuous pumping or disposable infusion.
Preferably, the animal is a rat.
Preferably, the perirenal fat is unilateral or bilateral perirenal fat of the animal.
Preferably, the blood pressure of the animal is measured weekly after administration until an animal model of hypertension is formed.
The invention only adopts a single intrarenal fat administration stimulation substance, and a stable hypertension animal model is obtained after 4-6 weeks, overcomes the defects of low molding rate, unstable model and the like of the traditional drug induction model (such as high-salt diet, high-fat diet, corticosteroids and the like), has the characteristics of 100 percent molding rate, adjustable blood pressure rising amplitude and the like, and can obtain a hypertension model with obviously raised blood pressure and a hypertension pre-stage model with slightly raised blood pressure by controlling the dosage of the stimulation substance such as complete Freund adjuvant. Compared with the existing spontaneous hypertension rat model, the invention avoids early-onset and inherent hypertension caused by genetic mutation and the like, is more similar to the clinical hypertension pathogenesis, and the grading type hypertension model of the invention cannot be realized by the spontaneous hypertension model.
Drawings
Fig. 1: and adopting capsaicin to continuously pump in the hypertension molding process.
Fig. 2: the hypertension molding process after the pumping is continued by using adenosine triphosphate.
Fig. 3: the hypertension molding process after the interleukin 6 is continuously pumped in is adopted.
Fig. 4: hypertension molding process after one injection of complete Freund's adjuvant 。
Wherein, saline is saline control, cap is capsaicin, ATP is adenosine triphosphate, IL-6 is interleukin 6, CFA indicates complete Freund's adjuvant SBP is systolic pressure, DBP is diastolic pressure, MAP is mean arterial pressure.
Detailed Description
In order to further describe the technical means and effects adopted by the present invention for achieving the intended purpose, the following detailed description will refer to the specific implementation, structure, characteristics and effects according to the present invention with reference to the accompanying drawings and preferred embodiments.
The blood pressure test method comprises the following steps: blood pressure was measured in the tail artery by a small animal blood pressure monitor ((Kent Scientific Corporation, CT, USA)). The rats were allowed to stand on a 28℃hotplate for 10-20 minutes and were measured by the instrument 8-12 times to obtain an average value.
Example 1
Continuous stimulation and induction hypertension model for capsaicin micropump in perirenal fat
(1) The front end metal tube of the microinjection pump (2006 model, alzet, USA) is connected with a PE50 tube, the length of the PE50 tube is 7-8cm, a notch is cut at the front end, and the front end of the PE50 tube stretches into the perirenal fat of the SD rat and is fixed;
(2) Continuously infusing capsaicin solution (capsaicin is purchased from Sigma Co., USA) with concentration of 0.1nmol/μl into perirenal fat site of SD rat by microinjection pump until hypertension model is formed, and administration time is 6 weeks in this example; the control group was filled with saline and the other groups were filled with capsaicin.
(3) Blood pressure measurements were taken after the initial administration and as shown in figure 1, it can be seen that blood pressure began to rise after 1 week of infusion, continued to rise for 6 weeks, and stabilized after week 7.
Example 2
(1) The front end metal tube of the microinjection pump (2006 model, alzet, USA) is connected with a PE50 tube, the length of the PE50 tube is 7-8cm, a notch is cut at the front end, and the front end of the PE50 tube stretches into the perirenal fat of the SD rat and is fixed;
(2) The perirenal fat sites of SD rats were continuously perfused with an ATP solution (ATP purchased from Sigma, usa) at a concentration of 3.5nmol/ul by a microinjection pump until a hypertensive model was formed, in this example for 6 weeks; control groups were filled with saline and others with ATP.
(3) Blood pressure measurements were taken after the initial administration and as shown in figure 2, it can be seen that blood pressure began to rise after 1 week of infusion, continued to rise for 6 weeks, and stabilized after week 7.
Example 3
(1) The front end metal tube of the microinjection pump (2006 model, alzet, USA) is connected with a PE50 tube, the length of the PE50 tube is 7-8cm, a notch is cut at the front end, and the front end of the PE50 tube stretches into the perirenal fat of the SD rat and is fixed;
(2) Continuous infusion of IL-6 solution (IL-6 was purchased from R & D company, USA) at a concentration of 1nmol/ul into perirenal fat sites of SD rats by means of a microinjection pump until a model of hypertension was formed, in this example for a period of 6 weeks; control groups were filled with saline and others with IL-6.
(3) Blood pressure measurements were taken after the initial administration and as shown in figure 3, it can be seen that blood pressure began to rise after 1 week of infusion, continued to rise for 6 weeks, and stabilized after week 7.
Example 4
(1) Complete Freund's adjuvant (1 mg/ml concentration; sigma, USA) was injected uniformly with a microinjector into bilateral perirenal fat in SD rats in single-sided doses of low, medium and high three dose groups, respectively, with 5ul (5 points/side fat; 1 μ/point;), 10ul for medium dose group and 15ul for high dose group.
(2) Blood pressure measurements were taken weekly after dosing and the results are shown in figure 2, as can be seen, blood pressure started to rise at 1 week of injection and stabilized at hypertensive state for 6 weeks of observation. Elevated blood pressure exhibits a dose-dependent relationship: the model blood pressure rise was most pronounced with 15ul administration, followed by 10ul, 5ul.
The present invention is not limited to the above embodiments, but is capable of modification and variation in detail, and other modifications and variations can be made by those skilled in the art without departing from the scope of the present invention.
Claims (4)
1. A preparation method of a hypertension animal model is characterized by comprising the following steps: injecting a chemical stimulus into perirenal fat of an animal, wherein the chemical stimulus is capsaicin, complete Freund's adjuvant, adenosine triphosphate or interleukin 6, and the animal is a rat.
2. The method for preparing an animal model of hypertension according to claim 1, characterized in that: the chemical stimulus is injected by continuous pumping or disposable injection.
3. The method for preparing an animal model of hypertension according to claim 1, characterized in that: the perirenal fat is unilateral or bilateral perirenal fat of the animal.
4. The method for preparing an animal model of hypertension according to claim 1, characterized in that: blood pressure was measured weekly for animals following dosing until an animal model of hypertension was established.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN202211244294.3A CN115443947B (en) | 2022-10-12 | 2022-10-12 | Preparation method of hypertension animal model |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202211244294.3A CN115443947B (en) | 2022-10-12 | 2022-10-12 | Preparation method of hypertension animal model |
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| CN115443947A CN115443947A (en) | 2022-12-09 |
| CN115443947B true CN115443947B (en) | 2023-11-21 |
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