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. 2022 Aug 12;20:117. doi: 10.1186/s12958-022-00991-6

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© The Author(s) 2022

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 2 — Effects of melatonin on AMPK/SIRT1 pathway dysregulation and mitochondrial dysfunction induced by high glucose. A The expression levels of SIRT1 and AMPK in rat testicular tissue detected by Western blot and the statistical graph of gray value analysis. B The expression levels of mitochondrial function-related proteins (mtTFA, ATPB and COXIV) in rat testicular tissue were detected by Western blot and their gray values were analyzed. C The expression levels of SIRT1, AMPK and mitochondrial function-related proteins in TM3 cells detected by Western blot and the statistical graphs of gray value analysis. D Acetylation level of total proteins in TM3 cells of each group was detected. E–F Protein levels of SIRT1, PGC-1α and NRF1 in cytoplasm and nucleus of TM3 cells were detected by Western blot and gray values of the protein bands were analyzed. G and J Mitochondrial membrane potential detection of TM3 cells by JC-1 probe and the ratio of J-aggregates to monomer (green fluorescence) was statistically analyzed. H and K Detection of the number and morphology of mitochondria in TM3 cells by MitoTracker and the percentage of cells with mitochondrial fragmentation (n = 150) was statistically analyzed. I and L Detection of mitochondrial reactive oxygen species in TM3 cells by MitoSOX and average fluorescence intensity analysis. Con, control group; DM, diabetes mellitus group; DM + INS, insulin treatment group of diabetic rats; DM + MT, melatonin treatment group of diabetic rats; NG, normal glucose control group; HG, high glucose treatment group; HG + MT, high glucose and melatonin treatment group; A-B, Data are expressed as fold change relative to Con; C, E–F, Data are expressed as fold change relative to NG. *, P < 0.05; #, P < 0.01. Bar: G-H, 15 μm; I, 50 μm