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發表期刊：Chinese Medicine

發表時間：2025

Abstract:

Background

Diabetic cardiomyopathy (DCM) is a cardiovascular complication, with cardiomyocyte senescence being a key pathological process. Cyclovirobuxine D (CVB-D), the active compound in Buxus sinica (Rehd. et Wils.) var. parvifolia M. Cheng. CVB-D has potentially promising diabetes-related cardiomyocyte senescence-mitigating effects. Nevertheless, the impact of CVB-D on inhibiting cardiomyocyte senescence has not been widely investigated and molecular mechanisms remain ambiguous.

Methods

A diabetic mouse model was established via a high-fat diet (HFD) combined with streptozotocin (STZ). Sirtuin 3 (SIRT3) knockout, SIRT3 overexpression, and ATP5O knockout mouse models were constructed through 4-week intravenous injections of AAV9-U6-SIRT3, AAV9-CMV-SIRT3, AAV9-U6-ATP5O, and their negative controls (AAV9-CMV-NC and AAV9-U6-NC). A primary mice cardiomyocytes (NMVMs) senescence model was developed using high palmitic acid and high glucose (PA/HG). Western blotting, reverse transcription-quantitative PCR (qRT-PCR), immunofluorescence, β-galactosidase staining and flow cytometry were performed to determine the protective role of CVB-D against cardiomyocyte senescence. The underlying mechanisms of CVB-D were investigated via molecular docking, coimmunoprecipitation (Co-IP), microscale thermophoresis (MST), surface plasmon resonance (SPR) binding assay, isothermal titration calorimetry (ITC) and LC–MS/MS analysis.

Results

CVB-D treatment improves mitochondrial dysfunction in DCM and thus alleviates the aging of cardiomyocytes in vitro and in vivo. And then, the results revealed that CVB-D can upregulate the acetylation level of ATP5O by upregulating the expression of SIRT3 to alleviate cardiomyocyte senescence. The results of Co-IP, MST, SPR, and ITC, among other experiments revealed that CVB-D plays a functional role through the SIRT3–ATP5O axis. Potential ATP5O acetylation sites by the LC–MS/MS analysis, we found that SIRT3 deacetylates the K162 site of ATP5O in primary mouse cardiomyocytes. Furthermore, transfection with a deacetylated or acetylated mimic plasmid containing ATP5O decreased or promoted mitochondrial damage, respectively. SIRT3 overexpression ameliorated DCM, whereas ATP5O knockout inhibited the protective effects of SIRT3 overexpression.

Conclusion

It is the first time that we confirm CVB-D ameliorating cardiomyocyte senescence in DCM by enhancing mitochondria dysfunction through activated SIRT3–ATP5O axis. It also suggests that CVB-D could be employed in the future to treat cardiomyocyte senescence caused by DCM.

https://doi.org/10.1186/s13020-025-01254-3