High glucose induces hippocampal neuron impairment through the SKP1/COX7C pathway: a potential mechanism for perimenopausal depression
Description
Perimenopause raises the risk and incidence of depression, whereas the underlying molecular mechanism remains unclear. Disturbed glucose regulation has been widely documented in depressive disorders, which renders the brain susceptible to various stresses such as estrogen depletion. However, whether and how glucose dysfunction regulates depression-like behaviors and neuronal damage in perimenopausal transition remains unexplored. Here, a prominent depressive phenotype was found in perimenopausal mice induced by the ovarian toxin 4‐vinylcyclohexene diepoxide (VCD). The VCD depression susceptible group (VCDSS) and the VCD depression resilient group (VCDRES) were determined using a ROC-based behavioral screening approach. We found that the hippocampus, a crucial region linked to depression, had hyperglycemia and mitochondrial abnormalities. Interestingly, oral administration of the SGLT2 inhibitor empagliflozin (EMPA) and intrahippocampal glucose infusion suggest a close relationship between hyperglycemia in the hippocampus and the susceptibility to depression. We verified that cytochrome c oxidase 7c (COX7C) downregulation is a potential cause of the high glucose-induced neuronal injury using proteomic screening and biochemical validations. High glucose causes COX7C to be ubiquitinated in a S-phase kinase associated protein 1 (SKP1)-dependent manner. According to these results, SKP1/COX7C represents a unique therapeutic target and a novel molecular route for treating perimenopausal depression. This data is the original data related to the study, including western blot, historical and fluorescence staining results.
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Mouse hippocampus tissues and cells were homogenized in RIPA buffer supplemented with a protease inhibitor complex. The protein concentration was evaluated by the BCA Protein Assay Kit. All samples were separated using SDS-PAGE gels and then placed onto a Millipore Immobilon-P Membrane in accordance with standard procedure. After blocking the membrane for 2 h with 5% skim milk, the primary antibody was incubated for the entire night at 4°C. The secondary antibody, peroxidase affinipure goat anti-rabbit or anti-mouse IgG, was exposed to ECL after being incubated for two hours at room temperature. Using Image J software, target protein band intensities were evaluated and converted to β-Actin to provide relative values. Perfused mice's uteri and brains were removed, and they were subsequently preserved for 24 h in 4% paraformaldehyde. The uterus was initially dehydrated with ethanol and then embedded in paraffin for hematoxylin and eosin (HE) staining. HE was used to stain each tissue after it had been divided into 5-µm-thick slices. Brain portions were dried in 30% sucrose solution, frozen in an OCT freezer at -80°C, and then sectioned using a cryostat into slices that were 15 μm thick for Nissl staining. Each tissue was stained with Nissl staining solution c. For immunofluorescence staining, after permeabilization with 0.3% Triton X-100 and 5% BSA in PBS, the slices were blocked in QuickBlock Immunostaining blocking solution before incubation with anti-COX7C antibody or anti-SKP1 antibody at 4°C overnight. After three washes with PBS, samples were incubated with TRITC-conjugated secondary antibody for 2 h at room temperature. The slices were washed three times with PBS and mounted with DAPI. A light microscope was used to capture the HE and Nissl images, and a confocal microscope was used to acquire the IHC images.
Institutions
- Shenyang Pharmaceutical UniversityLiaoning, Shenyang
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Funders
- Xingliao Talents ProgramGrant ID: XLYC1802007