Expression of ERK, JNK and p38 genes, and of proinflammatory cytokines TNF-α, IL-1β, and CRP in Mexican schoolchildren with distinct severity of dental fluorosis

Abstract

Objective: 1. To determine the expression of ERK, JNK and p38 genes, and of proinflammatory cytokines TNF-α, IL-1β, and CRP in Mexican schoolchildren with distinct severity of dental fluorosis.2. To compare the expression of ERK, JNK and p38 genes, and of proinflammatory cytokines TNF-α, IL-1β, and CRP according to the severity of dental fluorosis in the schoolchildren.

Methods: 120 schoolchildren will be screened for dental fluorosis by applying the severity index by Thylstrup and Fejerskov. Epithelial buccal cells will be collected via cytologic brushing, and preserved in RNAses inhibitory solution at -80°C. Extraction and purification of total RNA will be performed and the gene expression of ERK, JNK and p38 will be determined through RT-qPCR. The quantitative analysis of the gene expression will be completed through the 2^(-ΔΔCT) method. Non-stimulated saliva will be collected for the analysis of the protein expression of the proinflammatory cytokines TNF-α, IL-1β, and CRP in multiplex ELISA-sandwich-like assays.

Expected Results:The levels of expression for ERK and JNK genes are expected to be diminished, and to be elevated for p38 altogether with TNF-α, IL-1β and CRP in schoolchildren with higher severity of dental fluorosis; in comparison to the expression of these genes and proinflammatory cytokines in schoolchildren with lower or non-severity of dental fluorosis. This study will allow us to know the influence of dental fluorosis and inflammation on the expression of ERK, JNK and p38 genes.