Fig. 7: 8-AG reduces the expression of pro-inflammation genes, reduces the levels of pro-inflammatory cytokines, and affects the purine metabolome in F344 rat retinae.

Retina from young, water-treated aged, and 8-AG-treated aged rats were harvested, retinal lysates were used for qPCR, multiplex cytokine profiling using a flow-cytometer, and purine metabolome quantification using UPLC-MS/MS. a–f Transcript levels in the retinae for the following genes normalized with Gapdh level. N = 3. a Mt2a, b Adgre1, c Itgal, d Cd68, e Itgb2, and f Rt1-da. g–l Retinal cytokine/chemokine levels normalized by net retina weight and calibrated by standard curves. N = 6. g IL-33, h GM-CSF, i IL-28, j IL-1-alpha, k MCP-1, and l IL12p-70 (the 70 kDa complex of IL-12). m A brief illustration reminding the salvage and degradation of purine nucleosides. The tissue-damaging purines are in red background; 8-aminoguanine (8-AG) shown in light blue background, is a PNPase inhibitor. n–z Concentrations of retinal purine metabolites from young (black circle), water-treated aged (magenta square), and 8-AG-treated aged rats (blue triangle), normalized with the total protein in micrograms (ng of metabolite/µg of total protein). N = 4. n 8-AG; o Guanosine; p Adenosine; q Inosine; r 8-Aminoguanosine; s Guanine; t Adenine; u Hypoxanthine; v Xanthine; w 3’5’-cGMP; x 5’-GMP; y 3’5’-cAMP; z 5’-AMP. aa Dot plot of transcripts involved in the purine salvage and degradation pathway from young (black) and aged (magenta) rat retinae, using RNA-seq data. b Dot plot of transcripts involved in the purine salvage and degradation pathway from aged (magenta) and 8-AG-treated aged (blue) rat retinae, using the RNA-seq data. N = 3. Data and error bars are means±SDs. P values were calculated using One-way ANOVA for all the plots. ns stands for no significance.