CN-121970724-A - Construction method and application of plateau pulmonary artery high-pressure rat model

Abstract

The invention relates to a construction method and application of a plateau pulmonary artery high-pressure rat model, and belongs to the technical field of animal disease model construction. The method comprises the steps of firstly selecting healthy male SD rats with the age of 5-6 weeks and the weight of 220+/-10 g, adaptively breeding the healthy SD rats in a standard SPF-class animal room for 7 days, then dividing the rats into a healthy control group and a plateau hypoxia group, breeding the rats in the healthy control group in a normal-oxygen normal-temperature standard environment for 28 days, intermittently exposing the rats in the plateau hypoxia group in a multifunctional plateau environment experiment system for 28 days, wherein the intermittent exposure is specifically carried out by placing the rats in a simulated plateau hypoxia-low-pressure compound stress environment for 12 hours in a daily illumination period, and removing the rats in the normal-oxygen normal-temperature standard environment for recovery for 12 hours in a dark period. The rat model of the invention shows typical pulmonary artery high pressure characteristics, and various indexes are stable and repeatable, thus providing a reliable basis for research.

Inventors

• XING XIQIAN
• Xu Shuanglan
• YANG JIAO
• Qu Chongchang
• ZHANG XINTING
• WANG XIAOLAN
• Zheng Qinei
• CHEN YIDE

Assignees

• 云南大学附属医院

Dates

Publication Date

20260505

Application Date

20260325

Claims (5)

1. 1. The construction method of the plateau pulmonary artery high-pressure rat model is characterized by comprising the following steps of: Step one, preparing experimental animals: Healthy male SD rats with the age of 5-6 weeks and the weight of 220+/-10 g are selected and adaptively bred in standard SPF-class animal houses for 7 days; step two, grouping experimental animals and setting parameters: Rats were divided into healthy control and plateau hypoxia groups; the rats of the healthy control group are placed in a normal-oxygen normal-temperature standard environment for 28 days; The rats of the highland hypoxia group are placed in a cabin of a multifunctional highland environment experiment system for intermittent exposure for 28 days, and the environmental parameters of the cabin of the multifunctional highland environment experiment system are set as follows: The simulated altitude is 5500 meters; The oxygen concentration in the cabin is 10.0+/-0.5%; the temperature in the cabin is 16 ℃; The relative humidity in the cabin is 65%; Cabin pressure, -0.055 mPa; The intermittent exposure method comprises placing the rat in a multifunctional plateau environment experiment system cabin during 12 hours of illumination period, and removing the rat to normoxic normal temperature standard environment for recovery during the following 12 hours dark period; step three, evaluating a model; after the molding is finished, the rats are subjected to model evaluation, and the hemodynamic index, the right ventricular hypertrophy index and the lung histopathological index are detected during the evaluation.
2. 2. The method according to claim 1, wherein in the first step, the standard SPF-class animal room is set to a temperature of 22±2 ℃ and a humidity of 50±10%, and the standard SPF-class animal room is set to a 12-hour light cycle and a 12-hour dark cycle.
3. 3. The method for constructing a high-pressure rat model of the altitude pulmonary artery of claim 1, wherein in the second step, the normoxic normal temperature standard environment is specifically oxygen concentration of 20.9% and temperature of 22+ -2 ℃.
4. 4. The method according to claim 1, wherein in the third step, the plateau hypoxia group meets the following hemodynamic and histopathological indexes when evaluating, namely, the plateau pulmonary artery high pressure rat model is considered to be successfully constructed: (1) Compared with a healthy control group, the right ventricular systolic pressure of the rats in the plateau hypoxia group is increased, and the difference has statistical significance; (2) The right ventricular hypertrophy index shows that the rats with the plateau hypoxia group have obvious right ventricular hypertrophy, and the right ventricular hypertrophy index is obviously increased compared with the healthy control group; (3) Lung histopathological index: HE staining, namely obviously thickening the wall of the pulmonary arteriole in the altitude hypoxia group, narrowing the lumen and making inflammatory cells infiltrate around blood vessels compared with a healthy control group; EVG staining, namely, compared with a healthy control group, the plateau hypoxia group has the effects that the middle membrane of the pulmonary arteriole is thickened, the inner elastic membrane is distorted, layered or broken, and the area of a blood vessel cavity is obviously reduced; Masson staining: plateau hypoxia group collagen fiber deposition around pulmonary arterioles and alveolar spaces was significantly increased compared to healthy control group; Immunofluorescence the alpha-SMA fluorescence signal of the pulmonary arterioles of the rats in the plateau hypoxia group is obviously enhanced compared with that of the healthy control group, and meanwhile, the Col3a1 fluorescence signal is also obviously enhanced.
5. 5. The use of the altitude pulmonary artery high pressure rat model constructed by the altitude pulmonary artery high pressure rat model construction method according to any one of claims 1-4 in screening drugs for treating altitude pulmonary artery.

Description

Construction method and application of plateau pulmonary artery high-pressure rat model Technical Field The invention belongs to the technical field of experimental animal disease model construction, and particularly relates to a construction method and application of a plateau pulmonary artery high-pressure rat model. Background Plateau pulmonary arterial hypertension (High Altitude Pulmonary Hypertension, HAPH) is a chronic disease that results from prolonged exposure to the high altitude hypoxic environment, resulting in sustained constriction and remodeling of pulmonary vessels, which in turn leads to abnormally elevated pulmonary arterial pressure. HAPH is one of the main types of chronic altitude sickness, and is also the most common health problem among the plateau residents, and constitutes a serious threat to the health and life safety of plateau transferors and living beings. Early HAPH symptoms are not apparent, often delaying diagnosis or treatment, ultimately affecting the outcome of the intervention and patient prognosis. Systematic research HAPH is not only helpful for understanding disease mechanism and searching early intervention measures, but also has important significance for improving the health of plateau residents and guiding public health policy and special environmental medicine. Thus, the construction of a controllable and reproducible HAPH experimental animal model that is highly similar to the human plateau pulmonary hypertension pathology is critical for this disease study and intervention verification. Currently, in laboratory studies, researchers have developed various animal model construction methods for simulating pulmonary arterial hypertension, but these methods have respective significant limitations when applied to HAPH studies, and are briefly described below: 1. plain chemical induction model is the most common modeling mode of pulmonary artery high pressure model, mainly comprising: (1) The injection of monocrotaline (Monocrotaline, MCT) is metabolized in the liver following MCT injection, producing toxic pyrrole derivatives, causing injury to pulmonary artery endothelial cells and proliferation of pulmonary artery smooth muscle cells, thus mimicking some features of human pulmonary arterial hypertension, however, MCT-induced rats may die from pulmonary toxicity, venous embolic liver disease and myocarditis, rather than from pulmonary arterial hypertension, and its pathological changes do not fully represent a chronic hypoxic pathogenic process of human altitude pulmonary arterial hypertension; (2) Vascular endothelial growth factor receptor inhibitors (Sugen 5416) in combination with chronic hypoxia (SuHx) models, combined with chronic hypoxia (typically 10% O 2) exposure after administration of Sugen 5416, may induce more severe and persistent pulmonary vascular lesions similar to certain features of human pulmonary hypertension, however, the use of Sugen 5416 increases model complexity and the pharmacological effects of Sugen 5416 itself may have other effects on the pulmonary arterial hypertension pathology process, increasing difficulty in explaining the outcome; (3) The chronic hypoxia pulmonary hypertension is characterized in that endothelial cells are damaged due to chronic hypoxia, so that related contraction factors are unbalanced, pulmonary vasoconstriction reaction is increased, vascular remodeling is promoted, and compared with chemical induction, the chronic hypoxia model can better simulate the pathogenic cause of a highland hypoxia environment, and can be used for evaluating related pathological processes and intervention effects of hypoxia. However, the method has higher requirements on low-oxygen equipment and environmental control, the experimental period is long, and the model reproducibility is greatly influenced by the control precision of environmental parameters (such as oxygen concentration, temperature and humidity, pressure and the like). 2. The plateau induction model is a most direct construction mode of pulmonary artery high pressure model, namely, plain experimental animals (such as rats) are transported to a specific high-altitude area for long-term feeding, so that the plain experimental animals are naturally exposed to a composite plateau environment such as hypoxia, low temperature, low air pressure and the like, the method is closest to the occurrence and development of human HAPH in environmental etiology, can theoretically induce pathophysiological changes which are most similar to those of clinical patients, including pulmonary artery high pressure, right ventricular hypertrophy and pulmonary arteriole remodeling, but also has defects such as uncontrollable experimental environment, poor repeatability (natural fluctuation of parameters such as air pressure, oxygen partial pressure, temperature and the like, unstable results), high cost, low accessibility (large investment of experiments in the plain laboratory, great difficult