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ACCEPTED MANUSCRIPT with 670nm light did not have any altering effect on their expression (Figure 4B-F). The expression of Il-1β increased significantly in activated Müller cells, and 670nm treatment significantly reduced the expression level of this pro-inflammatory cytokine (P<0.05, Figure 4G), confirming our finding in in vivo (Figure 1A). 3.5 Mitochondrial function is improved by 670nm light following PD Cytochrome c oxidase (COX) is the terminal component of the electron transport chain in mitochondria and is the most likely photoacceptor of 670nm light (Desmet et al., 2006; Diaz, 2010; Karu, 1999). COX5a is one of 13 subunits of the COX enzyme in mammals. To understand whether COX participates in the effects of 670nm light on Müller cells, the expression of COX5a was measured in Müller cells. Low expression of COX5a was detected in the cytoplasm of Müller cells co-cultured with normal 661W cells (Figure 5B). Treatment with 670nm light of control Müller cells resulted in a significantly increased COX5a protein expression. In activated Müller cells, COX5a expression was significantly higher than control cells, and this expression further increased when activated cells were treated with 670nm (P<0.05, Figure 5B). Western blot analysis was used to quantify these changes (Figure 5C). Treatment with 670nm light increased the protein levels of COX5a in both control and activated Müller cells (P<0.05, Figure 5C, D). The increased expression of COX5a in control Müller cells following 670nm treatment confirmed the mechanistic effects of PBM. However, exposure to PD led to a higher expression of COX5a in Müller cells (P<0.05). The mitochondrial membrane potential (∆Ψm) is an important driver of ATP production and thus is a measure of mitochondrial function. The ∆Ψm significantly dropped in activated Müller cells compared to controls (P<0.05, Figure 5E). Treatment with 670nm 15 ACCEPTED MANUSCRIPTPDF Image | Photobiomodulation with 670 nm light ameliorates retinal
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