Research Paper Volume 16, Issue 3 pp 2438—2456

Interaction of Sp1 and Setd8 promotes vascular smooth muscle cells apoptosis by activating Mark4 in vascular calcification

Yun Li1,2, , Meijuan Cheng1,2, , Jingjing Jin1,2, , Dongxue Zhang1,2, , Shenglei Zhang1,2, , Yaling Bai1,2, , Jinsheng Xu1,2, ,

  • 1 Department of Nephrology, The Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, People’s Republic of China
  • 2 Hebei Clinical Research Center for Chronic Kidney Disease, Hebei Key Laboratory of Vascular Calcification in Kidney Disease, Shijiazhuang, People’s Republic of China

Received: October 10, 2023       Accepted: December 29, 2023       Published: February 1, 2024
How to Cite

Copyright: © 2024 Li et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


Vascular calcification (VC) is directly related to high mortality in chronic kidney disease (CKD), and cellular apoptosis of vascular smooth muscle cells (VSMCs) is a crucial process in the initiation of VC. Microtubule affinity-regulating kinase 4 (Mark4), known as a serine/threonine protein kinase, can induce cell apoptosis and autophagy by modulating Akt phosphorylation. However, the potential functions and molecular mechanisms of Mark4 in VSMCs apoptosis and calcification need to be further explored. Initially, our data indicated that the mRNA expression of Mark4 was prominently elevated in high phosphorus-stimulated human VSMCs compared with the other members in Marks. Consistently, Mark4 expression was found to be significantly increased in the calcified arteries of both CKD patients and rats. In vitro, silencing Mark4 suppressed apoptosis-specific marker expression by promoting Akt phosphorylation, finally attenuating VSMCs calcification induced by high phosphate. Mechanically, the transcription factor Sp1 was enriched in the Mark4 promoter region and modulated Mark4 transcription. Moreover, SET domain-containing protein 8 (Setd8) was proved to interact with Sp1 and jointly participated in the transcriptional regulation of Mark4. Finally, rescue experiments revealed that Setd8 contributed to VSMCs apoptosis and calcification by modulating Mark4 expression. In conclusion, these findings reveal that Mark4 is transcriptionally activated by Sp1, which is interacted with Setd8, to promote VSMCs calcification through Akt-mediated antiapoptotic effects, suggesting that Mark4 represents a potent and promising therapeutic target for VC in CKD.


CKD: Chronic kidney disease; VC: Vascular calcification; VSMCs: Vascular smooth muscle cells; Mark4: Microtubule affinity-regulating kinase4; Akt: Protein Kinase B; Sp1: Specific protein 1; Bcl-2: B-cell lymphoma-2; Bax: BCL2-Associated X; Setd8: SET domain-containing protein 8; caspase-3: Cysteinyl aspartate specific proteinase 3; Runx2: Runt-related transcription factor 2; GAPDH: Glyceraldehyde 3-phosphate dehydrogenase; DMEM: Dulbecco’s Modified Eagle Medium; FBS: Fetal Bovine Serum; BSA: Bovine Serum Albumin; RT-qPCR: Real-time-quantitative Polymerase Chain Reaction; ChIP: Chromatin immunoprecipitation.