Pulmonary arterial hypertension induced by a novel method: Twice-intraperitoneal injection of monocrotaline

Abstract

Pulmonary arterial hypertension (PAH) in humans manifests as a chronic process. However, PAH induced by high-dose monocrotaline (MCT) in animals occurs as a subacute process. To establish a chronic PAH model, rats were randomly divided into three groups, control (ctrl), single injection (SI), and twice injection (TI) groups. Rats in the SI group received a single intraperitoneal injection of 40 mg/kg MCT on day 0. Rats in the TI group received twice injections of 20 mg/kg MCT on days 0 and 7. Survival percentage, characteristic changes of pulmonary arterial variables, and right ventricular features were evaluated. Thirty-five days after the first MCT injection, survival percentage in TI group was higher than that in the SI group. The mean pulmonary arterial pressure (mPAP), right ventricular hypertrophy index (RVHI), pulmonary vascular remodeling, serum tumor necrosis factor α (TNFα), and interleukin-6 (IL-6) were higher either in SI or in TI 28 and 35 days after the first MCT injection. The rats in the SI and TI groups exhibited higher right ventricle end diastolic diameter (RVEDD) and lower adjusted pulmonary artery acceleration time (PAAT/HR), tricuspid annular plane systolic excursion (TAPSE), cardiac output (CO) and right ventricle fractional shortening (RVFS) when compared with controls. However, mPAP, RVHI, TAPSE, PAAT/HR, CO, TNFα, and IL-6 were lower and RVEDD were higher in the TI group than in the SI group. Pulmonary macrophage infiltration and right ventricle (RV) fibrosis were lower in TI than SI groups. The cardiomyocyte cross-sectional area and the beta myosin heavy chain (MYH7) mRNA level of RV were lower in TI than SI, whereas alpha myosin heavy chain (MYH6) was increased. These results show that two intraperitoneal injections of 20 mg/kg MCT with seven days interval could induce a model similar to chronic PAH with increased survival percentage in rats. Impact statement We demonstrated previously that a single intraperitoneal injection of 40 mg/kg MCT produced a subacute, not chronic, PAH model in rats, and the short survival periods of these rats did not represent adequately the chronic PAH seen in humans. To overcome this limitation, in this study, the single dose of 40 mg/kg MCT was divided into twice injections of 20 mg/kg with an interval of seven days. This modified administration of MCT produced an animal model much more similar to chronic PAH with prolonged survival and characteristic changes of structures and function in pulmonary arteries and right ventricles.

Preventive treatment with ginsenoside Rb1 ameliorates monocrotaline-induced pulmonary arterial hypertension in rats and involves store-operated calcium entry inhibition

“…The conventional MCT-induced PAH model has a short survival period, usually less than one month (Lachant et al 2018). Its subacute process cannot perfectly simulate the progressive development of human PH (Zhuang et al 2018). In addition, MCT-induced PAH appears to be preventable or curable by several agents, which is quite different from the case of human PH (Ruiter et al 2013;Colvin and Yeager 2014).…”

“…Its subacute process cannot perfectly simulate the progressive development of human PH (Zhuang et al. 2018 ). In addition, MCT-induced PAH appears to be preventable or curable by several agents, which is quite different from the case of human PH (Ruiter et al.…”

Transcriptomic analysis identifies Toll‐like and Nod‐like pathways and necroptosis in pulmonary arterial hypertension

“…PAH model in rats was induced by a single intraperitoneal injection of 40 mg/ kg MCT (Sigma-Aldrich) as described previously. 7 A total of 17 rats were used in this study: 12 rats were randomly assigned into 4 groups and treated with MCT (n = 3, each group) and five remaining rats served as control and treated with saline. Before killing, an effort was made to diminish suffering by intraperitoneal injection of 30 mg/kg sodium pentobarbital.…”

“…In addition to inflammatory cell infiltration, the inflammatory mediators, such as TNFα and IL-6, were elevated, with concomitantly increased pulmonary arterial remodelling parameters WT% and WA% in the progression of MCT-induced PAH. 7 The transcriptomic change during the PAH progression was investigated by using microarray. 8 Recently, high-throughput RNA sequencing (RNA-seq) has emerged as a more powerful alternative to microarray.…”

“…Consistently, our previous study has showed an elevated marker of macrophage infiltration. In addition to inflammatory cell infiltration, the inflammatory mediators, such as TNFα and IL‐6, were elevated, with concomitantly increased pulmonary arterial remodelling parameters WT% and WA% in the progression of MCT‐induced PAH 7 …”