CN-121970722-A - Construction method and application of animal blood stasis model

Abstract

The invention relates to the technical field of biology, in particular to a construction method and application of an animal blood stasis model. The invention successfully induces the animal blood stasis model by using the nitroglycerin. Experiments show that the treatment with the nitroglycerin can cause the zebra fish embryo to have blood stasis related phenotypes such as blood flow velocity blockage, heart red blood cell staining intensity reduction and blood coagulation factor and endothelial injury marker expression level increase, which indicates that the treatment with the nitroglycerin can successfully establish a zebra fish blood stasis model.

Inventors

• XIAO WEI
• LI JUAN
• XIE PAN
• LU MAN
• CAO LIANG

Assignees

• 江苏康缘药业股份有限公司

Dates

Publication Date

20260505

Application Date

20260211

Claims (10)

1. 1. Application of nitroglycerin in preparing animal blood stasis model is provided.
2. 2. The use according to claim 1, wherein the animal comprises zebra fish.
3. 3. The use according to claim 2, wherein the zebra fish comprises wild type AB strain zebra fish or Tg (gata 1: dsRed) red transgenic strain zebra fish.
4. 4. A method for constructing a blood stasis model, characterized in that nitroglycerin is administered to an animal.
5. 5. The method of claim 4, wherein administering nitroglycerin to the animal comprises culturing the animal with a medium comprising nitroglycerin.
6. 6. The construction method according to claim 5, wherein the concentration of nitroglycerin in the medium is 25 to 50. Mu.M.
7. 7. The method according to claim 4, wherein the period of shading of the feeding is 14h/10h.
8. 8. The method according to any one of claims 4 to 7, wherein the method comprises culturing zebra fish embryos in a medium containing nitroglycerin.
9. 9. The method of claim 8, wherein the zebra fish embryo is an embryo that develops to 2-3 dpf.
10. 10. The application of the animal blood stasis model obtained by the construction method of any one of claims 4-9 in screening blood circulation promoting and stasis removing medicines and/or antithrombotic medicines.

Description

Construction method and application of animal blood stasis model Technical Field The invention relates to the technical field of biology, in particular to a construction method and application of an animal blood stasis model. Background Blood stasis syndrome (Blood-stasis syndrome, BSS) is a basic syndrome commonly seen in traditional Chinese medicine in east Asia. The traditional Chinese medicine considers that the blood stasis syndrome is caused by qi stagnation, qi deficiency, cold coagulation, phlegm obstruction, blood heat, trauma and other reasons, such as blood circulation disorder or blood vessel obstruction, blood vessel obstruction or bleeding failing to dissipate in time. The main clinical manifestations are pain, lump, bleeding, dark purple complexion or cyanosis of lips and tongue, dry skin, excessive scales, thin and astringent pulse or intermittent pulse. The Zhang Zhongjing of the eastern Han period of doctors is based on the theory of the yellow emperor's internal channels, firstly, the disease name of blood stasis is determined, and a complete blood stasis syndrome differentiation and treatment system is created in the typhoid fever theory; the treatment methods of the Tang and Song period blood stasis syndrome and the blood circulation promoting and stasis removing are further developed in theory, prescription and medicine aspects, and the blood stasis theory is well supplemented. Modern medicine holds that blood stasis syndrome plays an important role in the onset and development of various diseases such as cardiovascular and cerebrovascular diseases, diabetes, tumor, inflammation, tissue hyperplasia, immune dysfunction and the like. It is essentially a microcirculation disturbance and blood coagulation increase, and is accompanied by various pathological changes such as denaturation, inflammation and pathological tumor. The mechanisms involved in these pathological processes include platelet activation, hyperclotting, abnormal haemorheological properties, and unbalanced expression of cell proliferation factors, inflammation-related factors and pro-fibrotic factors. Thrombosis (Thrombosis) is the process by which intravascular blood components coagulate to form solid masses (thrombi), a pathological phenomenon that results from a combination of vascular injury, stagnation of blood flow, and hypercoagulability of blood. Medical science researchers such as academy of China Chen Keji consider related documents, and combine modern clinical and experimental researches to consider that blood stasis can be divided into narrow and broad meaning that blood stasis refers to stagnation due to unsmooth blood circulation, broad meaning that blood stasis is caused by various causes of epidemic blood disorder or accumulated in pulse or overflowed outside pulse or thrombosis, and causes abnormality of blood related systems, so that blood functions, properties and components are changed, all of which are called blood stasis, and a series of clinical syndromes caused by blood stasis are called blood stasis syndrome. At present, the reported animal blood stasis models are mainly induced by a plurality of methods such as chemical substance induction, ice-water bath, exhaustion swimming and the like, and mainly comprise blood stasis models of higher animals such as mice, rats and the like. The modeling method is evaluated by observing whether the experimental animal has macroscopic phenotype and limb behaviours which accord with the traditional Chinese medicine symptoms or not and simultaneously utilizing blood rheology related indexes and the like. The blood stasis model constructed based on mammals such as mice, rats and the like can highly simulate pathophysiological changes of human blood stasis, has irreplaceable advantages in the aspects of deeply verifying the effectiveness of medicaments and clarifying the action mechanism of the medicaments, but has the same remarkable disadvantages, such as more experimental animals, longer modeling period and high overall experimental cost, so that the blood stasis animal model is only suitable for evaluating antithrombotic effects of a few medicaments or for discussing mechanisms, and is not suitable for large-scale screening of antithrombotic medicaments. For preliminary evaluation of drug activity and large-scale drug screening, which require rapid response, the efficiency is insufficient and the cost is too high. Therefore, the development of a novel blood stasis experimental animal model which is quick and economical has important significance. However, when animals such as rats, mice and the like are used for modeling, the number of the required animals is huge, the experimental operation is complex, the period is long, the experimental cost is high, the model repeatability is poor and the like, Zebra fish models exhibit a number of significant advantages over traditional mammalian models in drug screening studies. The zebra fish has small size, can be cultu