LncRNA‐ES3 inhibition by Bhlhe40 is involved in high glucose–induced calcification/senescence of vascular smooth muscle cells

Abstract Long noncoding RNAs (lncRNAs) have been investigated as novel regulatory molecules involved in diverse biological processes. Our previous study demonstrated that lncRNA‐ES3 is associated with the high glucose–induced calcification/senescence of human aortic vascular smooth muscle cells (HA‐VSMCs). However, the mechanism of lncRNA‐ES3 in vascular calcification/aging remained largely unknown. Here, we report that the expression of basic helix‐loop‐helix family member e40 (Bhlhe40) was decreased significantly in HA‐VSMCs treated with high glucose, whereas the expression of basic leucine zipper transcription factor (BATF) was increased. Overexpression of Bhlhe40 and inhibition of BATF alleviated calcification/senescence of HA‐VSMCs, as confirmed by Alizarin Red S staining and the presence of senescence‐associated β‐galactosidase–positive cells. Moreover, we identified that Bhlhe40 regulates lncRNA‐ES3 in HA‐VSMCs by binding to the promoter region of the lncRNA‐ES3 gene ( LINC00458 ). Upregulation or inhibition of lncRNA‐ES3 expression significantly promoted or reduced calcification/senescence of HA‐VSMCs, respectively. Additionally, we identified that lncRNA‐ES3 functions in this process by suppressing the expression of miR‐95‐5p, miR‐6776‐5p, miR‐3620‐5p, and miR‐4747‐5p. The results demonstrate that lncRNA‐ES3 triggers gene silencing of multiple miRNAs by binding to Bhlhe40, leading to calcification/senescence of VSMCs. Our findings suggest that pharmacological interventions targeting lncRNA‐ES3 may be therapeutically beneficial in ameliorating vascular calcification/aging.