B-type natriuretic peptide inhibits angiotensin II-induced proliferation and migration of pulmonary arterial smooth muscle cells

Abstract

Pulmonary vascular remodeling, characterized by disordered proliferation and migration of pulmonary arterial smooth muscle cells (PASMCs), is a pathognomonic feature of pulmonary arterial hypertension. Thus, pharmacologic strategy targeting on anti-proliferation and anti-migration of PASMCs may have therapeutic implications for PAH. Here we investigated the effects and underlying mechanisms of B-type natriuretic peptide (BNP) on angiotensin II (Ang II)-induced proliferation and migration of PASMCs. Proliferation and migration of PASMCs cultured from Wistar rats were induced by Ang II, with or without BNP treatment. In addition, potential underlying mechanisms including cell cycle progression, Ca(2+) overload, reactive oxygen species (ROS) production, signal transduction of MAPK and Akt, and the cGMP/PKG pathway were examined. We found that BNP inhibited Ang II-induced PASMCs proliferation and migration dose dependently. BNP could also arrest the cell cycle progression in the G0/G1-phase. In addition, BNP attenuated intracellular calcium overload caused by Ang II. Moreover, Ang II-induced ROS production was mitigated by BNP, with associated down-regulation of NAD(P)H oxidase 1 (Nox1) and reduced mitochondrial ROS production. Finally, Ang II-activated MAPKs and Akt were also counteracted by BNP. Of note, all these effects of BNP were abolished by a PKG inhibitor (Rp-8-Br-PET-cGMPS). In conclusion, BNP inhibits Ang II-induced PASMCs proliferation and migration. These effects are potentially mediated by decreased calcium influx, reduced ROS production by Nox1 and mitochondria, and down-regulation of MAPK and Akt signal transduction, through the cGMP/PKG pathway. Therefore, this study implicates that BNP may have a therapeutic role in pulmonary vascular remodeling.

Gamma-secretase Inhibitor Prevents Proliferation and Migration of Ductus Arteriosus Smooth Muscle Cells through the Notch3-HES1/2/5 Pathway

“…Culture media were changed every 3 days and primary cultures of two passages were used in the experiments. The purity of DASMC was confirmed under confocal microscope using immunofluorescence staining for α-smooth muscle actin (>95% of cells stained positive), with typical “hill and valley” appearance and lack of staining for vimentin, as we previously described 20. Serum starvation before experiments was achieved by 1% FBS for 48 h to synchronize cells into quiescence.…”

“…Then cells were trypsinized, centrifuged at 1,250 × g for 5 min, washed twice with cold PBS, and treated with RNase A (10 μg/ml). DNA was stained with propidium iodide (50 μg/ml) for 30 min at 37℃, and 5 × 10 3 cells were then analyzed by flow cytometry as we previously described 20.…”

Molecular Mechanisms for Regulating Postnatal Ductus Arteriosus Closure

“…Atrial natriuretic peptide has been shown to dilate rat DA in vivo [ 23 ]. Our recent study demonstrated that higher B-type natriuretic peptides (BNP) convey anti-remodeling effects in the pulmonary artery SMCs [ 46 ]. Furthermore, in the setting of neonatal intensive care unit, plasma levels of BNP are associated with poor response to indomethacin treatment in preterm infants with PDA [ 44 ].…”

Brain natriuretic peptide: Much more than a biomarker

“…Most of the information concerning the effects of BNP on ASM cell proliferation has been obtained from in vitro models. For example, BNP inhibited angiotensin IIinduced smooth muscle cell proliferation, likely mediated by a decreased calcium influx, reduced ROS production and Akt signal transduction[55].…”