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27 May 2018 : Laboratory Research  

Rho Kinase Type 1 (ROCK1) Promotes Lipopolysaccharide-induced Inflammation in Corneal Epithelial Cells by Activating Toll-Like Receptor 4 (TLR4)-Mediated Signaling

Jianying Gong1ABDE, Linan Guan1AG*, Pei Tian1BCD, Chao Li2CD, Yi Zhang1CD

DOI: 10.12659/MSM.907277

Med Sci Monit 2018; 24: LBR3514-3523

Abstract

BACKGROUND: Rho kinases (ROCKs) are the typical downstream effectors of RhoA, which can regulate corneal epithelial wound healing. In this study, the role of ROCK1 in lipopolysaccharide (LPS)-induced cornea inflammation was investigated.

MATERIAL AND METHODS: The expression of ROCK1 in human corneal epithelial cells (HCECs) was bilaterally modulated with ROCK1 expression vector and ROCK1 inhibitor Y-27632. The effects of ROCK1 modulation on the inflammatory response, cell viability, cell apoptosis, and cell cycle distribution were detected by ELISA assay, MTT assay, and flow cytometry, respectively. The pathways involved in the effect of ROCK1 in HCECs was preliminarily explained by detecting changes of TLR4-mediated NF-kB and ERK signaling using western blotting and electrophoretic mobility shift assays.

RESULTS: Overexpression of ROCK1 promoted LPS-induced production of IL-6, IL-8, IL-1β, and TNF-α, and the apoptotic process in HCECs. Augmented inflammation and apoptosis were associated with stronger activation of TLR4-mediated signal transduction; the phosphorylation of IkBa, JNK, ERK1/2, and p38, and nuclear translocation of NF-κB p65 induced by LPS were further increased by overexpression of ROCK1. Inhibition of ROCK1 function by Y-27632 blocked the effect of LPS on HCECs; both LPS-induced inflammation and apoptosis was alleviated by Y-27632, which was associated with suppression of TLR4-mediated NF-κB and ERK signaling.

CONCLUSIONS: LPS-induced inflammation and apoptosis in HCECs depended on the function of ROCK1, inhibition of which would attenuate impairments on cornea cells due to LPS.

Keywords: Corneal Diseases, Lipopolysaccharides, MAP Kinase Signaling System, NF-kappa B, Toll-Like Receptors

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750