Perinatal Research Facility, 13243 E. 23rd Ave., Mail Stop F441, Aurora, CO 80045.
Am J Physiol Lung Cell Mol Physiol. 2014 Feb;306(3):L299-308. doi: 10.1152/ajplung.00271.2013. Epub 2013 Dec 27.
Peroxisome proliferator-activated receptor-γ (PPARγ) and Rho-kinase (ROCK) regulate smooth muscle cell (SMC) proliferation and contribute to vascular remodeling in adult pulmonary hypertension. Whether these pathways interact to contribute to the development of vascular remodeling in persistent pulmonary hypertension of the newborn (PPHN) remains unknown. We hypothesized that ROCK-PPARγ interactions increase SMC proliferation resulting in vascular remodeling in experimental PPHN. Pulmonary artery SMCs (PASMCs) were harvested from fetal sheep after partial ligation of the ductus arteriosus in utero (PPHN) and controls. Cell counts were performed daily for 5 days with or without PPARγ agonists and ROCK inhibition. PPARγ and ROCK protein expression/activity were measured by Western blot in normal and PPHN PASMCs. We assessed PPARγ-ROCK interactions by studying the effect of ROCK activation on PPARγ activity and PPARγ inhibition (siRNA) on ROCK activity and PASMC proliferation. At baseline, PPHN PASMC cell number was increased by 38% above controls on day 5. ROCK protein expression/activity were increased by 25 and 34% and PPARγ protein/activity decreased by 40 and 50% in PPHN PASMC. ROCK inhibition and PPARγ activation restored PPHN PASMC growth to normal values. ROCK inhibition increased PPARγ activity by 50% in PPHN PASMC, restoring PPARγ activity to normal. In normal PASMCs, ROCK activation decreased PPARγ activity and PPARγ inhibition increased ROCK activity and cell proliferation, resulting in a PPHN hyperproliferative PASMC phenotype. PPARγ-ROCK interactions regulate SMC proliferation and contribute to increased PPHN PASMC proliferation and vascular remodeling in PPHN. Restoring normal PPARγ-ROCK signaling may prevent vascular remodeling and improve outcomes in PPHN.
过氧化物酶体增殖物激活受体-γ(PPARγ)和 Rho-激酶(ROCK)调节平滑肌细胞(SMC)增殖,并促进成人肺动脉高压中的血管重塑。这些途径是否相互作用以促进新生儿持续性肺动脉高压(PPHN)中的血管重塑尚不清楚。我们假设 ROCK-PPARγ 相互作用增加 SMC 增殖,导致实验性 PPHN 中的血管重塑。在宫内动脉导管部分结扎后从胎儿羊中收获肺动脉平滑肌细胞(PASMC),并分为 PPHN 组和对照组。在有无 PPARγ 激动剂和 ROCK 抑制的情况下,每天进行细胞计数 5 天。通过 Western blot 测量正常和 PPHN PASMC 中的 PPARγ 和 ROCK 蛋白表达/活性。通过研究 ROCK 激活对 PPARγ 活性的影响以及 PPARγ 抑制(siRNA)对 ROCK 活性和 PASMC 增殖的影响来评估 PPARγ-ROCK 相互作用。在基线时,PPHN PASMC 细胞数量在第 5 天比对照组增加 38%。PPHN PASMC 中的 ROCK 蛋白表达/活性增加了 25%和 34%,PPARγ 蛋白/活性降低了 40%和 50%。ROCK 抑制和 PPARγ 激活使 PPHN PASMC 的生长恢复正常。ROCK 抑制使 PPHN PASMC 中的 PPARγ 活性增加了 50%,恢复了正常的 PPARγ 活性。在正常 PASMC 中,ROCK 激活降低了 PPARγ 活性,PPARγ 抑制增加了 ROCK 活性和细胞增殖,导致 PPHN 中具有过度增殖表型的 PASMC。PPARγ-ROCK 相互作用调节 SMC 增殖,并导致 PPHN 中 PASMC 增殖和血管重塑增加。恢复正常的 PPARγ-ROCK 信号可能预防 PPHN 中的血管重塑并改善预后。