Hypertrophy and prolonged DNA synthesis in smooth muscle cells characterize pulmonary arterial wall thickening after monocrotaline pyrrole administration to rats.

Abstract

Monocrotaline pyrrole (MCTP) is a highly reactive pneumotoxic metabolite of the pyrrolizidine alkaloid plant toxin monocrotaline. When administered to rats, it causes a delayed and progressive lung injury, vascular remodeling, and pulmonary hypertension. Structural remodeling consists of endothelial cell swelling followed by increased thickness of the vascular media in small pulmonary arteries and muscularization of normally nonmuscular arteries. Experiments were performed to characterize DNA synthesis and cell proliferation in vascular smooth muscle cells (VSMCs) after MCTP and to determine their relationship to changes in the thickness of the arterial medial layer of pulmonary resistance vessels. Male Sprague-Dawley rats were treated with MCTP (3.5 mg/kg, intravenously) or its vehicle (dimethylformamide). To label cells actively synthesizing DNA, rats were given the thymidine analog, bromodeoxyuridine (BrdU), 3 times by intraperitoneal injection during the 24 hr preceding euthanasia. Using immunohistochemistry, BrdU incorporation was quantified as a ratio of labeled nuclei to total nuclei. Within 5 days after MCTP administration, the thickness of the medial smooth muscle layer in arteries 60-250 microm in diameter was increased, prior to evidence of right heart hypertrophy. BrdU incorporation by VSMCs in pulmonary arteries was not different in vehicle- and MCTP-treated rats for the first 48 hr after treatment. However, MCTP caused a significant increase in DNA synthesis in VSMC on days 3-8 in arteries up to 250 microm in diameter. Although increased DNA synthesis precedes cell proliferation, the relative number of medial VSMCs did not increase over 8 days, suggesting that hypertrophy alone was responsible for the increased thickness of the arterial media. These results demonstrate that MCTP causes thickening of the media of pulmonary vessels through VSMC hypertrophy and that the prolonged DNA synthesis that accompanies VSMC hypertrophy is not followed by proliferation.