ROCK inhibitor fasudil reduces the expression of inflammatory factors in LPS-induced rat pulmonary microvascular endothelial cells via ROS/NF-κB pathway

Abstract

Background Inflammation plays a major role in the pulmonary artery hypertension (PAH) and the acute lung injury (ALI) diseases. The common feature of these complications is the dysfunction of pulmonary microvascular endothelial cells (PMVECs). Fasudil, the only Rho kinase (ROCK) inhibitor used in clinic, has been proved to be the most promising new drug for the treatment of PAH, with some anti-inflammatory activity. Therefore, in the present study, the effect of fasudil on lipopolysaccharide (LPS)-induced inflammatory injury in rat PMVECs was investigated. Methods LPS was used to make inflammatory injury model of rat PMVECs. Thereafter, the mRNA and protein expression of pro-inflammatory factors was evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) assay respectively. Intracellular reactive oxygen species (ROS) levels were measured by the confocal laser scanning system. The activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and the content of malondialdehyde (MDA) were determined by using commercial kits according to the manufacturer’s instructions. Western blot assay was used to detect the protein expression of nuclear factor kappa B (NF-κB) p65. Results Fasudil effectively prevented inflammatory injury induced by LPS, which is manifested by the decrease of pro-inflammatory cytokines interleukin-6 (IL-6) and monocyte chenotactic protein-1 (MCP-1). Meanwhile, fasudil dramatically reduced the levels of ROS and MDA, and also elevated the activities of SOD and GSH-Px. Furthermore, the nuclear translocation of NF-κB p65 induced by LPS was also suppressed by fasudil. Additionally, the ROS scavengers N-Acetylcysteine (N-Ace) was also found to inhibit the nuclear translocation of NF-κB and the mRNA expression of IL-6 and MCP-1 induced by LPS, which suggested that ROS was essential for the nuclear translocation of NF-κB. Conclusions The present study revealed that fasudil reduced the expression of inflammatory factors, alleviated the inflammatory and oxidative damage induced by LPS in rat PMVECs via ROS-NF-κB signaling pathway.

Regulation of Microtubule Stability in Pulmonary Microvascular Endothelial Cells in Rats with Severe Acute Pancreatitis: Qingyi Decoction is a Potential CDK5 Inhibitor

“… 38 The ROCK inhibitor fasudil reduces the expression of inflammatory factors in LPS-induced rat lung microvascular endothelial cells via the ROS/NF-κB pathway. 39 Therefore, PMVECs are essential to preventing the development of SAP-associated ALI.…”

Conditional deficiency of Rho‐associated kinases disrupts endothelial cell junctions and impairs respiratory function in adult mice

“…Regarding the first possibility, previous reports have shown that activation of Rho‐ROCK signaling leads to disruption of cell–cell adhesion through cell tension generated by actomyosin activation in cultured human umbilical vein endothelial cells (HUVEC) [ 38 , 39 ]. Furthermore, ROCK inhibitors have been reported to suppress the increase in vascular permeability that occurs in response to infection or inflammatory agents in vivo [ 40 , 41 ]. Thus, activation of ROCK is thought to be involved in the increase in vascular permeability.…”

Inhibitors of Mitochondrial Dynamics Mediated by Dynamin-Related Protein 1 in Pulmonary Arterial Hypertension

“…Fasudil is one of its typical inhibitors, which has been approved in clinical treatment. In PAH rats, Fasudil and its derivative, Fasudil dichloroacetate, could prevent PAH development by reversing right ventricular systolic pressure (RVSP) and right ventricle hypertrophy index (RVHI), preventing pulmonary vascular remodeling, and also RV hypertrophy and fibrosis, and decreasing inflammatory factor expression ( Wenshu et al, 2018 ; Qi et al, 2019 ; Liu et al, 2022 ). In clinical treatment, Fasudil revealed a similar effect in PAH patients that decreased pulmonary vascular resistance PVR and PAP, as well as increased mean cardiac output and mixed venous oxygen saturation ( Li et al, 2009 ; Kojonazarov et al, 2012 ; Fukumoto et al, 2013 ; Jiang et al, 2014 ; Xiao et al, 2015 ).…”