Pulmonary arterial hypertension (PAH) is a progressive disease caused by a variety of pulmonary and/or cardiac disorders. PAH is commonly characterized by an increase in the pulmonary vascular resistance or in the right ventricular systolic pressure (RVSP). A rise in RVSP induces the right ventricular (RV) hypertrophy, and RV failure, and may eventually lead to premature death. It is well known that a single subcutaneous administration of a pyrrolizidine alkaloid monocrotaline (MCT) to rats causes cardiovascular and pulmonary disorders similar to those in patients with PAH 1,2) including severe RV hypertrophy 3,4) and RV failure. [5][6][7][8] Despite extensive studies, serial events of MCT-induced functional changes in the development and progression of RV failure have not been fully elucidated.The present study was designed to elucidate pathophysiological and pharmacological alterations of the RV function of MCT administered rats. To examine time-matched alterations of the RV function of MCT administered animals, we focused on the time course of study on cardiac function by a noninvasive echocardiography in rats.
MATERIALS AND METHODS
AnimalsMale Wistar rats (SLC, Hamamatsu, Japan) weighting 190-210 g were used in the present study. Rats were randomly selected to receive a subcutaneous injection of either monocrotaline (MCT, 60 mg/kg) or an equal volume of saline (CON). The animals were conditioned according to Guide for the Care and Use of Laboratory Animals as published by the U.S. National Institutes of Health (NIH Publication No. 85-23, revised 1996). The protocol of this study was approved by the Committee of Animal Use and Welfare of Tokyo University of Pharmacy and Life Sciences.Echocardiography Transthoracic echocardiography was performed on rats according to the method described previously, 9) in which experimental conditions employed, including appropriate anesthesia, were extensively examined. In the present study, rats were evaluated 2, 4, 6, and 8 weeks after administration of either MCT (2w-, 4w-, 6w-, and 8w-MCT rats) or vehicle (2w-, 4w-, 6w-, and 8w-CON rats, respectively).The animals were anesthetized with 40 mg/kg intraperitoneally (i.p.) pentobarbital sodium, and then their hairs on the chest were shaved before examination. Two-dimensional and Doppler imaging were performed by using ProSound 5500 R (Aloka, Tokyo, Japan) with a 10-MHz transducer. The transthoracic echocardiographic probe was placed to obtain long-axis and apical four-chamber views. To evaluate right ventricular (RV) function, cardiac output (CO) and stroke volume (SV) at the pulmonary artery and the RV Tei index were measured. After determination of the pulmonary arterial flow, heart rate (HR), velocity time integral (VTI), and pulmonary arterial diameter (PAD) were measured by the long-axis view. The ratios of CO and SV to body weight (BW) were calculated according to the following equations (CO and SV indices (COI and SVI), respectively) 9,10) : COIϭ(PAD/2) 2 ϫpϫVTIϫHR/BW, SVIϭCO/(HRϫBW). In addition, to characterize the pu...