Fig. 2: Hcy induces mitochondrial dysfunction in the hippocampus of rats.

A, B Representative TEM images of mitochondria during Hcy treatment. The red “m” in the images represented mitochondria (n = 5 for each group, 6 serial sections per rat). C Relative mtDNA/nDNA ratio in the hippocampus of Con and HHcy rats. D, E Western blotting of p-AMPKα1/AMPKα, m-PGC1α/PGC1α, NRF1, and TFAM protein abundance in the hippocampus of Con and HHcy rats. F, G Red arrows indicated damaged mitochondria. All mitochondria were divided into 4 levels according to the degree of damage from mild to severe. The scoring criteria referred to [74]. H, I Western blotting measured the expression of LC3B, P62, PINK1, PARKIN, and TOMM40 in the hippocampus of Con and Hcy groups. J Representative confocal fluorescence micrographs from the CA1 region of the hippocampus stimulated with or without Hcy and subjected to immunofluorescence labeling of TOMM40 (green) and LC3B (red). K The representative line scans were from the LC3B puncta colocalizing with TOMM40. L Relative PINK1 activity in the hippocampus of Con and HHcy rats. M–Q Relative ATP, MDA, MitoSOX level, and SOD activity in the hippocampus of Con and HHcy rats. R, S Immunoblot of COX5A, SDHB, UQCRC2, and ATP5A in the hippocampus of Con and HHcy rats. T The relative activity of Sirt1 in the hippocampus between Con and Hcy groups was measured using the Sirt1 activity assay. U RT-qPCR verification of Sirt1. Gapdh normalized the data. V, W Immunoblot analysis of p-Sirt1/Sirt1 in the hippocampus of Con and HHcy rats. X–Z NAD⁺, NADH, and NAD⁺/NADH levels of the hippocampus in rats between Con and Hcy groups were measured by a related kit (n = 5 for each group). Data were presented as mean ± SEM. Unpaired t-test was used to analyze the data (*P < 0.05, **P < 0.01, ***P < 0.001; n = 6 for each group).