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Julia Tau, Priscila Novaes, Monique Matsuda, Paulo HN Saldiva, Alejandro Berra; Diesel Exhaust Particles selectively induce both proinflamatories cytokines and mucins production in Cornea and Conjunctiva Human Cell Lines. Invest. Ophthalmol. Vis. Sci. 2012;53(14):2319.
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© ARVO (1962-2015); The Authors (2016-present)
To evaluate the effect of Diesel Exhaust Particles (DEP) at various concentrations (10, 50, 100 and 500 ug/ml) on the viability, migration, secretion of cytokines (IL-6, IL-8 and TNF-α) and the production of mucins (MUC1, MUC5AC and MUC16) in human epithelial cells of conjunctiva (IOBA-NHC)and cornea (HCLE).
as viability assay we performed cell count with 4% trypan blue. To assess cell migration we used the method of the wound healing. The determination of pro-inflammatory cytokines was done with ELISA kit. Finally, the mucin expression levels were quantified by Real Time PCR using the Rotor Gene equipment.
a dose-dependent decrease in viability and cell migration (p <0.001) was obtained in both IOBA-NHC and HCLE exposed to DEP. IOBA-NHC showed a dose-dependent increase in the release of IL-6 (p = 0.014 for DEP 500 ug/ml) and a dose dependent decrease in the release of IL-8 (p = 0.023 for DEP 100 ug/ml, p <0.001 for DEP 500 ug/ml). HCLE stimulated with DEP showed a dose-dependent statistically significant decrease (p <0.001) in IL-8 secretion. But, was not detected IL-6 secretion. Also, was not detected release of TNF-α or cornea or conjunctiva. IOBA-NHC stimulated by DEP showed a increased expression of mucins, it was statistically significant for DEP 100 ug/ml for MUC1, p <0.001, for MUC5AC, p = 0.049 and MUC16, p <0.001. In contrast, HCLE stimulated by DEP showed a reduced expression of mucins, it was statistically significant for DEP 10 and 100 ug/ml for MUC1 (p = 0.016, p = 0.002) and MUC16 (p = 0.020, p = 0.003), for MUC5AC the decrease was statistically significant (p = 0.032) only for 100 ug/ml of DEP.
DEP has a dose-dependent effect in decreasing the viability and cell migration and secretion of IL-8 in both cells of cornea and conjunctiva. Instead, DEP increased the secretion of IL-6 only in cells of the conjunctiva and had no effect on the secretion of TNF-α. Finally, DEP stimulate mucin production in human conjunctival cells, whereas in human corneal cells inhibit the expression of these mucins.
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