Effects of ROCK Inhibitors on Apoptosis of Corneal Endothelial Cells in CMV-Positive Posner-Schlossman Syndrome Patients.

ABSTRACT: Purpose:To examine the role of aqueous tumor necrosis factor ? (TNF-?)-RhoA-Rho kinase (ROCK) signaling in cytomegalovirus (CMV)-induced apoptosis and the barrier function of cultured human corneal endothelial cells (hCECs) in CMV-positive Posner-Schlossman syndrome (CMV+/PSS) patients. Methods:Aqueous levels of TNF-?, IL-8, IL-10, and several other cytokines in 19 CMV+/PSS patients and 20 healthy control subjects were quantitated using a multiplex assay. The expression of active RhoA in hCECs post-CMV infection was determined using western blotting (WB). The expression levels of TNF-? and nuclear factor kappa B (NF-?B) in CMV-infected hCECs were examined by immunocytochemistry (ICC) and WB with and without ROCK inhibitors. The apoptotic rate and barrier integrity in CMV-infected hCECs were also examined. Results:The expression levels of TNF-?, monocyte chemoattractant protein-1 (MCP-1), IL-8, and IL-10 were upregulated in the aqueous humor of CMV+/PSS patients, and among these upregulated cytokines aqueous TNF-? was negatively correlated with the number of corneal endothelial cells. In CMV-infected hCECs, upregulation of TNF-? and NF-?B was determined by WB and ICC. In hCECs, CMV infection induced apoptosis and significantly impaired cell-cell contacts, effects that were attenuated by treatment with a ROCK inhibitor. Conclusions:Aqueous TNF-? was upregulated in CMV+/PSS patients, which may have triggered corneal endothelial cell loss. Modulation of TNF-?, including its downstream Rho-ROCK signaling, could serve as a novel treatment modality for corneal endothelial cell loss in CMV+/PSS patients.

SUBMITTER: Igarashi N 

PROVIDER: S-EPMC7441372 | biostudies-literature | 2020 Aug

REPOSITORIES: biostudies-literature