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. 2016 Jan 19;17(1):117. doi: 10.3390/ijms17010117

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© 2016 by the authors; licensee MDPI, Basel, Switzerland.

This article is an open access article distributed under the terms and conditions of the Creative Commons by Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).

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Both the PI3K/Akt/eNOS pathway and the ERK1/2 pathways were involved in the neuroprotective effects of CHR20/21. Pre-incubation of LY294002, Akt VIII inhibitor, and PD98059 at a dose of 10 µM, respectively blocked the neuroprotective effects of CHR20 (A) and CHR21 (B) in RGC-5 cells. MTT activity was determined by MTT assay. Each experiment was performed in triplicate. ^## p < 0.01 vs. SNP group; ** p < 0.01 vs. CHR20/CHR21 + SNP group; ^& p < 0.05, ^&& p < 0.01 vs. CHR20/CHR21 + PD group, or CHR20/CHR21 + LY group, respectively; (C) SNP inhibited the phosphorylations of Akt, eNOS and ERK1/2, respectively. And LY294002 and PD98059 reversed the effects of CHR21. The density of each lane was presented as mean ± standard deviation for three individual experiments. * p < 0.05 vs. control group, ^&& p < 0.01 vs. SNP group, ^## p < 0.01 vs. CHR21 + SNP group. Blots were quantified using Image J software. Similar results were obtained with CHR20 (data not shown).

Both the PI3K/Akt/eNOS pathway and the ERK1/2 pathways were involved in the neuroprotective effects of CHR20/21. Pre-incubation of LY294002, Akt VIII inhibitor, and PD98059 at a dose of 10 µM, respectively blocked the neuroprotective effects of CHR20 (A) and CHR21 (B) in RGC-5 cells. MTT activity was determined by MTT assay. Each experiment was performed in triplicate. ^## p < 0.01 vs. SNP group; ** p < 0.01 vs. CHR20/CHR21 + SNP group; ^& p < 0.05, ^&& p < 0.01 vs. CHR20/CHR21 + PD group, or CHR20/CHR21 + LY group, respectively; (C) SNP inhibited the phosphorylations of Akt, eNOS and ERK1/2, respectively. And LY294002 and PD98059 reversed the effects of CHR21. The density of each lane was presented as mean ± standard deviation for three individual experiments. * p < 0.05 vs. control group, ^&& p < 0.01 vs. SNP group, ^## p < 0.01 vs. CHR21 + SNP group. Blots were quantified using Image J software. Similar results were obtained with CHR20 (data not shown).