Abstract
Endothelial dysfunction, a central hallmark of cardiovascular pathogenesis in diabetes mellitus, is characterized by impaired endothelial nitric oxide synthase (eNOS) and NO bioavailability. However, the underlying mechanisms remain unclear. Here in this study, we aimed to identify the role of calmodulin (CaM) in diabetic eNOS dysfunction. Human umbilical vein endothelial cells and murine endothelial progenitor cells (EPCs) treated with high glucose (HG) exhibited downregulated CaM mRNA/protein and vascular endothelial growth factor (VEGF) expression with impeded eNOS phosphorylation and cell migration/tube formation. These perturbations were reduplicated in CALM1-knockdown cells but prevented in CALM1-overexpressing cells. EPCs from type 2 diabetes animals behaved similarly to HG-treated normal EPCs, which could be rescued by CALM1-gene transduction. Consistently, diabetic animals displayed impaired eNOS phosphorylation, endothelium-dependent dilation, and CaM expression in the aorta, as well as deficient physical interaction of CaM and eNOS in the gastrocnemius. Local CALM1 gene delivery into a diabetic mouse ischemic hindlimb improved the blunted limb blood perfusion and gastrocnemius angiogenesis, and foot injuries. Diabetic patients showed insufficient foot microvascular autoregulation, eNOS phosphorylation, and NO production with downregulated CaM expression in the arterial endothelium, and abnormal CALM1 transcription in genome-wide sequencing analysis. Therefore, our findings demonstrated that downregulated CaM expression is responsible for endothelium dysfunction and angiogenesis impairment in diabetes, and provided a novel mechanism and target to protect against diabetic endothelial injury.
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Acknowledgements
We thank Dr. Jin-huan Gao from Xuanwu Hospital, Capital Medical University for technical support in the establishment of hind limb ischemia model. This study was supported by the National Natural Science Foundation of China (81570206 and 81970197), and the Scientific Research Key Program of Beijing Municipal Commission of Education (KZ201710025023).
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TTL, HHX, and ZJL performed the research and wrote the manuscript. HPZ, ZXZ and HTZ prepared patient samples and collected information. ZQW, JYX, QL, YM and HJY provided research material and techniques and contributed to data interpretation. DLL directed the project, wrote, reviewed, and edited the manuscript. DLL is the guarantor of this project and, as such, has full access to all the data in the study and take responsibility for the integrity of the data and the accuracy of the data analysis.
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Liu, Tt., Xu, Hh., Liu, Zj. et al. Downregulated calmodulin expression contributes to endothelial cell impairment in diabetes. Acta Pharmacol Sin 44, 2492–2503 (2023). https://doi.org/10.1038/s41401-023-01127-1
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DOI: https://doi.org/10.1038/s41401-023-01127-1
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