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Construction method of sepsis compound animal model

A construction method and animal model technology, applied in veterinary surgery, medical science, veterinary instruments, etc., can solve problems that are not conducive to the protection and welfare of experimental animals, accidental death, short distance, etc., and achieve the effect of saving the number of experimental animals

Pending Publication Date: 2020-11-13
SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI +1
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  • Claims
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Problems solved by technology

Although common carotid artery and jugular vein intubation are commonly used for blood collection, the location of the intubation is close to the main organs, which can easily cause accidental death, and it is difficult to achieve blood collection in rats in a free-moving state
At present, animal plasma samples for sepsis pharmacokinetic research are mainly collected from the orbital blood of rats, and in order to ensure data quality, usually a mouse can only collect blood samples at one time point, resulting in a large loss of experimental animals
[0005] To sum up, the number of animals lost in the pharmacokinetic study by routine blood collection on the CLP model is large, which not only increases the cost of the experiment, but also is not conducive to the protection and welfare of experimental animals.

Method used

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  • Construction method of sepsis compound animal model
  • Construction method of sepsis compound animal model
  • Construction method of sepsis compound animal model

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Embodiment 1 femoral artery cannulation (FAC) model preparation

[0073] After rats were anesthetized (8ml / kg) with chloral hydrate solution (50mg / ml), use an electric shaver to shave the hair on the rat's thigh (ventral side), groin and near the tail, and wipe the shaved area with disinfectant alcohol. parts. The rats were placed in a supine position with their heads facing forward on a constant temperature (37°C) heating pad, their limbs were pasted and fixed with medical adhesive tape, and the operation area was exposed to the field of view of the operation microscope. Such as Picture 1-1 As shown, an incision of about 1.0 cm was made along the parallel direction of the groin, the femoral artery was exposed after blunt dissection, and a section of surgical silk (about 5 cm) for ligation was pre-placed under the proximal end of the arterial vessel. Clamp the proximal end of the blood vessel with an arterial clip, make an oblique opening on the distal end of the bloo...

Embodiment 2

[0074] Example 2 Femoral artery cannulation (FAC) combined with cecal ligation and puncture (CLP) sepsis pharmacokinetic rat composite model preparation

[0075] Forty-eight hours after the rats were successfully prepared with femoral artery cannulation, the sepsis model was prepared by cecum ligation and puncture (CLP), and the operation method was improved CLP operation. After successful anesthesia, the rats were fixed in the supine position on the operating table, and the skin of the abdominal operation area was smeared with amyl iodine twice, and then the hair on the abdomen was shaved off with a razor blade; a longitudinal incision of about 2 cm was made along the midline of the abdomen, Separate the skin and muscular layer, free the cecum, and ligate with 4.0 silk thread at 30-50% from the top of the cecum to the lower part of the ileocecal valve ( diagram 2-1 ); and puncture the cecum twice with a 16G puncture needle to form intestinal fistula ( Figure 2-2 ), and squ...

Embodiment 3

[0076] The animal mortality evaluation of embodiment 3FAC combined CLP composite model

[0077] The 28-day mortality rate was the main clinical outcome index of sepsis, and the animal observation period was 7 days. 150 rats were randomly divided into Sham group (n=30), CLP group (n=30), FAC+CLP group (n=30), CLP+XBJ group (n=30), FAC+CLP+XBJ group (n=30), XBJ in the above groups stands for Xuebijing. Each group was treated as follows: (1) Sham group: 4 mL / kg of normal saline was injected intravenously at 2, 12, 24, 36, 48, and 60 hours after operation. (2) CLP group: 4 mL / kg of normal saline was injected intravenously at 2, 12, 24, 36, 48, and 60 hours after CLP operation. (3) CLP+FAC group: FAC rats were injected with normal saline 4 mL / kg intravenously at 2, 12, 24, 36, 48, and 60 hours after CLP operation. (4) CLP+XBJ group: Xuebijing 4 mL / kg was injected intravenously at 2, 12, 24, 36, 48, and 60 hours after CLP operation. (5) CLP+FAC+XBJ group: 2, 12, 24, 36, 48, and ...

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Abstract

The invention relates to the field of animal models for basic scientific research, in particular to a construction method of a sepsis composite animal model, and the construction method comprises femoral artery intubation and cecum ligation puncture. The femoral artery intubation and cecum ligation puncture are combined for the first time to construct the sepsis composite animal model, the sepsiscomposite animal model is successfully applied to pharmacokinetic study of anti-sepsis drugs on model rats, continuous blood sample collection on the same rat can be achieved, the death rate of overall animals is not affected, and the number of experimental animals is greatly reduced. Meanwhile, reliable model animal pharmacokinetic data can be obtained, and important reference significance is achieved for drug research in related fields.

Description

technical field [0001] The invention relates to the field of animal models for basic scientific research, in particular to a method for constructing a composite animal model of sepsis. Background technique [0002] Sepsis is an organ dysfunction syndrome caused by a systemic host reaction caused by infection. It is a dangerous condition with a high fatality rate (25-50%) and poor prognosis. It has become a difficulty faced by modern critical care medicine. The conventional treatment of sepsis (including septic shock) mainly relies on timely and reasonable antibiotic anti-infection treatment and regulation of abnormal reactions of patients. However, there is still no effective chemical drug for the latter, and the development of new drugs is still in progress. The traditional Chinese medicine injection Xuebijing is currently the only Class II new drug approved for the treatment of sepsis in China (in 2004). About 600,000 sepsis patients in China use Xuebijing every year. Clin...

Claims

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Application Information

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IPC IPC(8): A61D1/00
CPCA61D1/00
Inventor 李川程晨杜飞飞徐方王凤清余玄董凯张桂萍姚小青
Owner SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI
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