FIGURE 7.

Cytokine regulation by Grx1 in rMC-1 cells. Cytokine expression in the culture medium of rMC-1 cells overexpressing Grx1 in normal glucose medium was measured via a multiplex cytokine Luminex enzyme-linked immunosorbent assay (ELISA), and VEGF protein was analyzed from the same samples of medium using a VEGF ELISA. Release of TNF-α, IFN-γ, IL-1β, and IL-10 into the medium was not detected (panel a). IL-6 and VEGF were both detected in the medium, but only IL-6 was induced by the overexpression of Grx1 in Müller cells (panel a). rMC-1 Müller cells overexpressing Grx1 with Ad-Grx at an m.o.i. of 10 were cultured for 24 h in normal glucose medium. Medium from the cells was then collected, concentrated 3-fold, and tested in an IL-6 ELISA. Medium from rMC-1 Müller cells overexpressing Grx1 showed a 3.7-fold (±0.5) increase in IL-6 expression over cells transfected with a control empty vector construct (Ad-Empty) or untransfected (0) (panel b)(n = 6) *, p = 0.003. Treatment with recombinant IL-6 induced expression of Grx by 1.5-fold (±0.2) and ICAM-1 by 2.1-fold (±0.4) *, p ≤ 0.03 (n = 8) (panel c).