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A modeling method for migraine model rats with wind-heat syndrome

A technology for migraine and wind-heat syndrome, which is applied to the modeling field of wind-heat migraine model rats. good effect

Active Publication Date: 2022-01-28
赵永烈 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of these models ignore the coincidence with TCM syndrome types, and the modeling factors are single
At present, animal models related to wind-heat syndrome in traditional Chinese medicine are used in the research of upper respiratory tract infection and pneumonia. The modeling methods include chemical drugs to induce cough, fever, and local inflammation. These modeling methods are based on the mechanism of Western medicine. , simulating pathological changes, and there is a big gap with the "wind-heat syndrome" in traditional Chinese medicine's etiology meaning. It can be said that there is no animal model of wind-heat syndrome in line with the etiology and pathogenesis theory of traditional Chinese medicine.

Method used

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  • A modeling method for migraine model rats with wind-heat syndrome
  • A modeling method for migraine model rats with wind-heat syndrome
  • A modeling method for migraine model rats with wind-heat syndrome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Acquisition and identification of migraine model of wind-heat syndrome

[0039] 1. Experimental consumables

[0040] 1.1 Experimental animals: 60 SPF grade male SD rats, each weighing 180-220g, were purchased from Beijing Weitong Lihua Experimental Animal Research Center, license number: SCXK (Beijing) 2016-0006, and the animals were kept in clean grade Animal room, indoor temperature 22±2°C, relative humidity 30-40%, 5 rats per cage, drinking tap water sterilized by high-pressure steam, and feeding ordinary rat pellets by themselves. The experiments were carried out after the animals were adaptively fed for 3 days after purchase.

[0041] 1.2 Experimental drugs: gastrodin tablets, produced by Shanghai Modern Hasen (Shangqiu) Pharmaceutical Co., Ltd., daidzein tablets (puerarin tablets), produced by Shanxi Aoer Pharmaceutical Co., Ltd. The daily dose of Gastrodin Tablets for adults is 150-300 mg. In this experiment, 300 mg is used. The dose for rats is calculated as...

Embodiment 2

[0052] The steps of detection of CGRP protein immunopositive reactants in brain slices of each group are as follows:

[0053] 1. Immunohistochemical staining:

[0054] (1) Transfer the brain tissue slices to a 24-well plate, suck off the antifreeze, and rinse with 0.01MPBS for 3×5min;

[0055] (2) Add prepared 1% Triton-100, 500 μl per well, incubate at 37°C for 1 hour on a constant temperature shaker;

[0056] (3) Aspirate 1% Triton-100, add 0.01MPBS, rinse 3×5min;

[0057] (4) Add the prepared 3% H 2 o 2 , 500 μl per well, place at room temperature in the dark for 15 minutes;

[0058] (5) Rinse with pure water for 2×5min, and rinse with 0.01MPBS for 10min;

[0059] (6) Add 10% goat serum blocking solution, 500 μl per well, incubate at 37°C for 1 hour on a constant temperature shaker;

[0060] (7) Aspirate the blocked goat serum without rinsing, and directly add the CGRP primary antibody diluted in antibody diluent (dilution ratio 1:1000), 500 μl per well, first incubat...

Embodiment 3

[0070] Detection of RAMP1 and CRLR protein expression in the thalamus, trigeminal ganglion, and spinal nucleus of the trigeminal nerve, the steps are as follows:

[0071] 1. Weston-blot process:

[0072] 1.1 Protein extraction:

[0073] (1) Mark and weigh the centrifuge tube, and place it on the centrifuge tube rack. Take the tissue out of the -70°C refrigerator, quickly pick the tissue in the original tube into the prepared specimen centrifuge tube with clean tweezers on the ice tray, weigh it again, and calculate the difference;

[0074] (2) Add the lysate with a difference of 10 times (μl) into the centrifuge tubes of each specimen, and homogenize with a hand-held electric homogenizer. The liquid in the centrifuge tube is transferred to the sample centrifuge tube;

[0075] (3) Put the sample centrifuge tube in a centrifuge, centrifuge at 4°C, 12000 rpm, and centrifuge for 15 minutes;

[0076] (4) Carefully draw the supernatant into another centrifuge tube and store at -...

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Abstract

The embodiment of the invention discloses a modeling method for an animal model of migraine due to wind-heat syndrome. Swimming at normal temperature is designed to cause animal fatigue; hot air is blown in a constant temperature climate box to simulate the external wind-heat environment; and nitroglycerin is injected on the back of the neck to induce migraine-like Pathological state. In this way, an animal model of wind-heat syndrome migraine was obtained and evaluated. The results show that this method can successfully reproduce the animal model of wind-heat syndrome migraine, and the replication method has good stability and repeatability, which is in line with the guidance of the theory of traditional Chinese medicine. The present invention also relates to the animal model obtained by the modeling method.

Description

technical field [0001] The invention relates to the technical field of traditional Chinese medicine evaluation, in particular to a modeling method for migraine model rats with wind-heat syndrome. Background technique [0002] Migraine is characterized by unilateral moderate to severe throbbing headache as the main symptom, sometimes alternately bilateral or even bilateral in severe cases, often accompanied by photophobia, phonophobia, nausea, vomiting and other symptoms. It has the characteristics of long course of disease, easy to repeat, lingering and difficult to heal. The pathogenesis of migraine is still not very clear. The mainstream theory believes that migraine is closely related to trigeminal neurovascular theory, cortical spreading depression, genetic factors and biochemical factors. It is a very common primary neurological disease in clinical practice. . [0003] Nitroglycerin, as a NO donor, can induce headache attacks in migraine patients. The rodent model es...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01K67/02
CPCA01K67/02A01K2227/105A01K2267/0356A01K2207/20A01K2207/35
Inventor 赵永烈张庆美
Owner 赵永烈
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