Ripasudil Attenuates Lipopolysaccharide (LPS)-Mediated Apoptosis and Inflammation in Pulmonary Microvascular Endothelial Cells via ROCK2/eNOS Signaling

Medical Science Monitor : International Medical Journal of Experimental and Clinical Research
Jianxin YangZhongjun Zheng

Abstract

BACKGROUND Microvascular endothelial inflammation and apoptosis are responsible for septic acute lung injury (ALI). Ripasudil is a novel Rho/Rho kinase (ROCK) inhibitor which shows therapeutic effects on several vascular diseases. The aim of this study was to investigate the protective effects and correlated molecular mechanisms of ripasudil on lipopolysaccharide- induced inflammation and apoptosis of pulmonary microvascular endothelial cells (PMVECs). MATERIAL AND METHODS Cultured PMVECs were exposed to lipopolysaccharide (LPS). Ripasudil at various concentrations was used to treat the cells. Several cells were also co-administrated with the endothelial nitric oxide synthase (eNOS) inhibitor Nω-Nitro-L-arginine methyl ester hydrochloride (L-NAME). Cell viability was assessed by MTT assay. Terminal dUTP transferase nick-end labeling (TUNEL) assay was used to detect the apoptosis. The colorimetric method was used to measure the activity of eNOS and ROCK2. Protein phosphorylation and expression were assessed by Western blotting. RESULTS Ripasudil attenuated the LPS-induced inflammation and apoptosis in PMVECs, which was reversed by L-NAME. Ripasudil suppressed ROCK2 activity and further increased the eNOS activity. Ripasudil trea...Continue Reading

Citations

Aug 5, 2020·Investigative Ophthalmology & Visual Science·Nozomi IgarashiMakoto Aihara
Jun 3, 2020·Journal of Molecular Neuroscience : MN·Lei LiuLijian Pan
Jul 30, 2020·Pharmacological Research : the Official Journal of the Italian Pharmacological Society·Graziella Ribeiro de SousaMaría Sol Brassesco

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Methods Mentioned

BETA
GTPase
enzyme-linked immunosorbent assay
nuclear translocation
bronchoalveolar lavage
nucleotide exchange

Software Mentioned

Image J
SPSS

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