Establishment and Validation of a Rat Model of Pulmonary Arterial Hypertension Associated with Pulmonary Fibrosis

Qian Jiang; Dongmei Wen; Fang Peng; Wei Hong; Xin Fu; Yuanhui Xiong; Chongyuan Ren; Xuanyi Li; Dansha Zhou; Jian Wang; Yuqin Chen

doi:10.3791/67747

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Medicine

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JoVE Journal Medicine

Establishment and Validation of a Rat Model of Pulmonary Arterial Hypertension Associated with Pulmonary Fibrosis

肺动脉高压与肺纤维化相关大鼠模型的建立与验证

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07:11 min

May 23, 2025

DOI: 10.3791/67747-v

Qian Jiang*¹, Dongmei Wen*¹, Fang Peng*², Wei Hong³, Xin Fu³, Yuanhui Xiong¹, Chongyuan Ren¹, Xuanyi Li¹, Dansha Zhou^1,4, Jian Wang^1,4, Yuqin Chen¹

¹State Key Laboratory of Respiratory Diseases, National Center for Respiratory Medicine, Guangdong Key Laboratory of Vascular Diseases, National Clinical Research Center for Respiratory Diseases, Guangzhou Institute of Respiratory Health,the First Affiliated Hospital of Guangzhou Medical University, ²Department of Critical Care Medicine,The Third Affiliated Hospital of Guangzhou Medical University, ³GMU-GIBH Joint School of Life Sciences,Guangzhou Medical University, ⁴Guangzhou laboratory,Guangzhou International Bio Island

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Overview

This study introduces a method for inducing pulmonary arterial hypertension associated with pulmonary fibrosis (PF-PH) in a rat model through bleomycin injection. It also details a step-by-step approach for validating this animal model.

Key Study Components

Area of Science

Pulmonary hypertension
Animal models
Pathophysiology

Background

Group three pulmonary hypertension is poorly understood.
Recent findings link metabolic dysregulation to pulmonary hypertension.
Research focuses on vascular remodeling and therapeutic targets.
Animal models are crucial for studying disease mechanisms.

Purpose of Study

To validate a rat model for PF-PH.
To explore the molecular mechanisms of pulmonary hypertension.
To assess hemodynamic changes and lung function.

Methods Used

Intubation of the trachea for bleomycin injection.
Measurement of right ventricular systolic pressure (RVSP).
Histological analysis for collagen accumulation.
Assessment of lung function via forced vital capacity (FVC).

Main Results

Increased RVSP and right ventricle to left ventricle ratio in the model group.
Histological staining showed greater collagen accumulation in the model group.
Reduced FVC and dynamic lung compliance in the model group.
Higher hydroxyproline concentration indicating increased collagen content.

Conclusions

The bleomycin-induced PF-PH rat model effectively mimics human disease.
Significant hemodynamic and histological changes were observed.
This model can be used for further research into therapeutic targets.

Frequently Asked Questions

What is the significance of the PF-PH rat model?

The PF-PH rat model helps in understanding the mechanisms of pulmonary hypertension associated with pulmonary fibrosis.

How is bleomycin administered in this study?

Bleomycin is injected into the airway of the rat using a syringe and cannula.

What measurements are taken to assess lung function?

Forced vital capacity (FVC) and dynamic lung compliance are measured to assess lung function.

What histological changes are observed in the model?

Increased collagen accumulation around pulmonary arteries is observed in the model group.

What does RVSP indicate in this study?

Right ventricular systolic pressure (RVSP) indicates the pressure in the right ventricle, reflecting pulmonary vascular resistance.

How does this study contribute to pulmonary hypertension research?

It provides a validated animal model for studying the pathogenesis and potential treatments for PF-PH.

本文介绍了通过将博来霉素注射到气道中来诱导肺动脉高压与肺纤维化相关的肺动脉高压（PF-PH）大鼠模型的方法。我们还提供了一种分步方法来验证这种动物模型。

本研究探讨了第三组肺动脉高压的发病机制，重点关注肺纤维化病例的分子机制。与肺动脉高压相比，间质性肺病相关肺动脉高压的机制仍知之甚少。该领域的最新发展包括逆转血管重塑的功效。发现癌症样内皮表型和代谢发现，将氨基酸和脂质失调与肺动脉高压的发病机制联系起来。批量和单细胞 I-A 测序代谢学和功能测定，例如烦人病例耐药性测试，是绘制肺动脉高压、异质性和确定治疗靶点的关键技术。

[导师]首先，使用PE-50管插管大鼠的气管，插入三厘米的深度，这是从声门到管末端的距离。使用带有 26 号针头的 1 毫升注射器，抽取 0.2 毫升博来霉素，并通过插管将其作为单剂量注射到气道中。将大鼠置于仰卧位，以确保博来霉素均匀分布。轻轻握住并慢慢旋转大鼠，以 30 度至 45 度角交替两侧。保持每次旋转 10 到 15 秒，然后重复三到四次。打开 PFT 软件并检查设备是否被识别。现在打开主控仪表开关并确保每个信号放大器开关已关闭。要设置气流，请打开吸入气流调节螺母并将吸气设置为五。然后打开呼气气流调节螺母，将慢呼气设置为负四。将压力值设置为 60。按FRC流量、流量、高流量、肺压的顺序校准仪器，误差小于5%。麻醉大鼠后，沿颈部中央切开皮肤。将肌肉逐层分开，露出颈气管的上部。在气管软骨环之间做一个水平切口。快速将金属插管插入气管切口，确保正确定位，并用四根零丝线缝合线固定。将大鼠放入PFT动物隔间中，并将气管插管连接到仪器外部的气流通道接口。单击开始按钮并检查用力肺活量或 FVC 和动态肺顺应性。为了检测右心室收缩压或RVSP，将麻醉大鼠的腹部向上固定在实验台上。用手术剪刀在颈部右侧做一个约四厘米长的切口。用显微手术镊子轻轻分离颈外静脉，确保隔离约一厘米的长度。在分离静脉的远端和近端插入两根手术线。用四根零丝线结扎颈外静脉远端。轻轻提起近端手术线并将其放在远端静脉壁上。用小剪刀做一个约 0.3 毫米的小切口。将PE-50管插入颈外静脉后，用四根零丝线将PE-50管和静脉结扎在一起，以防止渗血。观察记录仪上的静脉压波形。将导管缓慢推入右心房，推进至右心室，显示右心室压力波形。当压力稳定时，记录右心室压一分钟，并保存数据以备分析。开胸手术后，用镊子取出大鼠完整的心脏。用剪刀剪掉甲骨文和心脏附近的结缔组织。沿着肺动脉切开右心室的游离壁。该组织是右心室，其余组织是左心室加室间隔。分离后，用滤纸引流右心室和左心室加脑室内隔表面。在计算右心室和左心室加室间隔的重量比之前，分别用分析天平称量它们的重量比。该图说明了在博来霉素诱导的PF PH大鼠模型中观察到的血流动力学变化，肺血管重塑和肺功能受损。与对照组相比，模型组的右心室收缩压和右心室与左心室加间隔比值显著升高。组织学染色显示模型组肺动脉周围蓝色染色区域较多，表明胶原蛋白积累较多。与对照组相比，模型组的用力肺活量和动态肺顺应性显着降低。模型组肺动脉加速时间与射血时间之比、三尖瓣环平面收缩偏移、右心室面积分数变化、心输出量显著降低。模型组羟脯氨酸浓度显著升高，表明肺组织中胶原蛋白含量增加。对照组和模型组的肝脏和肾脏重量差异无统计学意义。

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