Corticosteroids effects on LPS-induced rat inflammatory keratocyte cell model

Abstract

PurposeCorticosteroids are efficient anti-inflammation treatments. However, there are still arguments on whether it should be used in keratitis. This study was to observe the effect of corticosteroids on keratocytes both in normal condition and inflammation status in vitro.MethodsRat keratocytes were cultured and used for examination. 10 μg/ml lipopolysaccharide (LPS) was used to establish the inflammatory keratocyte cell model, and prednisolone acetate (PA), dexamethasone (Dex) and fluorometholone (Flu) were used as corticosteroids treatments. 5 d-growth curve and cell viabilities were assayed by CCK8, and cell morphologies and migration rate were studied. TNF-α, IL-6 and IL-1β levels were examined by ELISA. Western blotting was used to quantified type VI collagen (Col VI) and matrix metalloproteinase 9 (MMP9) expressions, and immunofluorescence staining assays of Col I and Col VI were carried out.ResultsIn normal condition, proliferation and migration of keratocytes were slightly influenced in PA, Dex and Flu groups. The secretion of Col I and Col VI was suppressed and MMP9 expression increased in corticosteroids groups. But no significant difference was seen in TNF-α, IL-6 and IL-1β expression levels. In inflammatory status, TNF-α, IL-6 and MMP9 levels increased in LPS group, while they significantly decreased in corticosteroids groups. Although keratocytes viabilities and migration were slightly affected in 24 h, no significant differences were seen between LPS group and corticosteroids groups in 5-d proliferation. Col I and Col VI secretion in LPS-keratocytes was maintained with corticosteroids treatments.ConclusionsCorticosteroids showed lightly effects on keratocytes proliferation and migration, but it successfully decreased TNF-α, IL-6 level and maintained the secretion of and Col I and Col VI, while suppressed the expression of MMP9 in LPS-induced keratocytes. PA was suggested to use in early stage of keratitis clinical treatment.

Fluorometholone modulates gene expression of ocular surface mucins

“…; Yan et al. ). Fluorometholone for ophthalmic use is available as 0.1% suspension which is equivalent to 2600 μ m concentration.…”

“…; Yan et al. ). To test whether the effect of fluorometholone on mucin gene expression was glucocorticoid receptor‐mediated, human conjunctival and corneal epithelial cells were exposed to fluorometholone (100 n m ) alone and in the presence of a glucocorticoid receptor antagonist, mifepristone (10 μ m ), for 24 hr.…”

Dexamethasone can attenuate the pulmonary inflammatory response via regulation of the lncH19/miR-324-3p cascade

“…NF-κΒ is a transcription factor regulating many of the cytokine and adhesion molecule genes expressed in allergic in ammation, cytokine including colonystimulating factor and TNF-α, adhesion molecules including intercellular adhesion molecule (ICAM)-1 and vascular cell adhesionmolecule-1 (VCAM-1) [5]. Dex is a synthetic glucocorticoid that exerts anti-in ammatory effects by inhibiting NF-κΒ signaling pathway [6]. However, the molecular mechanism of Dex in pulmonary in ammatory diseases remains unclear.…”

Effect of Tempol, a Membrane-Permeable Free Radical Scavenger, on In Vitro Model of Eye Inflammation on Rabbit Corneal Cells

“…This could result in the reduction of corneal stroma and successive scarring creation. 4 Several studies reported that corneal injury during inflammatory condition is connected principally to the polymorphonuclear neutrophil (PMN) infiltration in the corneal tissue with the relief of proteolytic enzymes, arachidonic acid, and reactive oxygen species (ROS). 5,6 In keratitis, inflammation frequently persists and lapses long after the bacteria were removed, causing numerous injuries to the structure and cornea function.…”