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Updated: Aug 24, 2026
Calcification of Vascular Smooth Muscle Cells and Imaging of Aortic Calcification and Inflammation
Published on: May 31, 2016
Exosomal ALKBH5 Alleviates Vascular Calcification by Suppressing Cell Apoptosis via m6A-Modified GSDME
Guian Xu1, Qingman Li1, Lijie Zhu1
1Department of Cardiology, Central China Fuwai Hospital, Zhengzhou University, Zhengzhou, China.
Abstract:
This study aimed to explore the function and regulatory mechanism of ALKBH5 in the progression of coronary artery calcification. Human aortic vascular smooth muscle cells (HA-VSMCs) were treated with inorganic phosphate (Pi) and exosomes derived from bone marrow mesenchymal stem cell (BMSC) carrying ALKBH5, a GSDME overexpression vector or si-GSDME. The morphology and size of the exosomes were assessed using nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM). Calcium deposition was measured using Alizarin red staining and cell pyroptosis was evaluated using Hoechst 33342/PI staining. The association between ALKBH5 and m6A modifications was confirmed by methylated-RNA immunoprecipitation assay (MeRIP) and dot blot assays. The expression levels of ALKBH5 and GSDME were quantified by quantitative real-time polymerase chain reaction (qRT-PCR), and protein levels were quantified by western blot. BMSCs-derived exosomes reduced calcium deposition and cell pyroptosis in Pi-treated HA-VSMCs. Exosomes containing ALKBH5 overexpression inhibited high mobility group box 1 (HMGB1) and cell apoptosis, thereby promoting vascular calcification, whereas ALKBH5 knockdown in exosomes exerted the opposite effect on calcification development. Additionally, ALKBH5 was found to regulate the m6A modification of GSDME. Overexpression of GSDME reversed the effects of ALKBH5 in exosomes on HMGB1 expression and cell apoptosis. Exosomal ALKBH5 mitigated HMGB1 expression and cell pyroptosis by modulating the m6A modification of GSDME, thus influencing the progression of coronary artery calcification.
Insights
Exosomes carrying ALKBH5 from bone marrow mesenchymal stem cells (BMSCs) reduce vascular calcification by inhibiting pyroptosis. ALKBH5 regulates GSDME m6A modification, impacting HMGB1 expression and cell apoptosis in coronary artery disease.
Area of Science:
- Cardiovascular Biology
- Molecular Medicine
- Cellular Biology
Background:
- Coronary artery calcification (CAC) is a hallmark of atherosclerosis.
- The role of specific RNA modifications and exosomal contents in CAC progression remains incompletely understood.
Purpose of the Study:
- To investigate the function and regulatory mechanism of AlkB homolog 5 (ALKBH5) in coronary artery calcification.
- To elucidate the role of ALKBH5-modified exosomes derived from bone marrow mesenchymal stem cells (BMSCs) in regulating vascular smooth muscle cell pyroptosis and calcification.
Main Methods:
- Human aortic vascular smooth muscle cells (HA-VSMCs) were treated with inorganic phosphate (Pi) and engineered exosomes.
- Exosome characterization (NTA, TEM), calcium deposition assay (Alizarin red), pyroptosis assessment (Hoechst 33342/PI staining), m6A modification analysis (MeRIP, dot blot), gene expression (qRT-PCR), and protein analysis (western blot) were performed.
Main Results:
- BMSCs-derived exosomes attenuated Pi-induced calcium deposition and pyroptosis in HA-VSMCs.
- Exosomal ALKBH5 overexpression inhibited high mobility group box 1 (HMGB1) and cell apoptosis, promoting vascular calcification, while ALKBH5 knockdown had opposing effects.
- ALKBH5 was confirmed to regulate the m6A modification of Gasdermin E (GSDME), and GSDME overexpression reversed the effects of exosomal ALKBH5 on HMGB1 and apoptosis.
Conclusions:
- Exosomal ALKBH5 mitigates HMGB1 expression and cell pyroptosis by modulating GSDME m6A modification.
- This mechanism influences the progression of coronary artery calcification, highlighting exosomal ALKBH5 as a potential therapeutic target.
