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Both high glucose and phosphate overload promote senescence-associated calcification of vascular muscle cells

  • Nephrology - Original Paper
  • Published:
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Abstract

Purpose

The NAD+-dependent deacetylase, sirtuin 1 (SIRT1), plays an important role in vascular calcification induced by high glucose and/or high phosphate levels. However, the mechanism by which SIRT1 regulates this process is still not fully understood. Thus, this study aimed to determine the role of high glucose and phosphate in vascular calcification and the molecular mechanisms underlying SIRT1 regulation.

Methods

Vascular smooth muscle cells (VSMCs) were cultured under normal, high phosphate, and/or high-glucose conditions for 9 days. Alizarin red staining and calcification content analyses were used to determine calcium deposition. VSMC senescence was detected by β-galactosidase (SA-β-Gal) staining and p21 expression.

Results

Mouse VSMCs exposed to high phosphate and high glucose in vitro showed increased calcification, which was correlated with the induction of cell senescence, as confirmed by the increased SA-β-galactosidase activity and p21 expression. SRT1720, an activator of SIRT1, inhibits p65 acetylation, the nuclear factor-κ-gene binding (NF-κB) pathway, and VSMC transdifferentiation, prevents senescence and reactive oxygen species (ROS) production, and reduces vascular calcification. In contrast, sirtinol, an inhibitor of SIRT1, increases p65 acetylation, activates the NF-κB pathway, induces vascular smooth muscle cell transdifferentiation and senescence, and promotes vascular calcification.

Conclusions

High glucose and high phosphate levels induce senescence and vascular calcification in VSMCs, and the combined effect of high glucose and phosphate can inhibit SIRT1 expression. SIRT1 inhibits vascular smooth muscle cell senescence and osteogenic differentiation by inhibiting NF-κB activity, thereby inhibiting vascular calcification.

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Abbreviations

CKD:

Chronic kidney disease

DM:

Diabetes mellitus

VSMCs:

Vascular smooth muscle cells

NF-κB:

Nuclear factor-κ-gene binding

α-SMA:

Alpha smooth muscle actin

SM22α:

Smooth muscle 22 alpha

RUNX2:

Runt-related transcription factor-2

Pit-1:

Type III sodium‑dependent phosphate cotransporter-1

BMP2:

Bone morphogenetic protein 2

VC:

Vascular calcification

SA-β-Gal:

Senescence-associated-β-galactosidase

SIRT1:

Sirtuin 1

AcP65:

Acetylated-p65

ROS:

Reactive oxygen species

ESRD:

End stage renal disease

VC:

Vascular calcification

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Funding

This work was supported by grants from the Natural Science Foundation of Anhui Province (No. 2008085MH244) and the National Natural Science Foundation of China in the Second Hospital of Anhui Medical University (No. 2020GMFY04).

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Authors and Affiliations

  1. Department of Nephrology, The Second Affiliated Hospital of Anhui Medical University, Hefei, China

    Mingming Zhang, Tianyu Li, Yuying Zhang, Xuerong Wang & Deguang Wang

  2. Department of Biochemistry and Molecular Biology, Anhui Medical University, Hefei, China

    Zhenzhen Tu, Yuying Zhang, Yang Feng, Fan He, Mingyue Ni & Haisheng Zhou

  3. The Center for Scientific Research of Anhui Medical University, Hefei, China

    Dandan Zang & Haisheng Zhou

  4. Department of Clinical Laboratory, Affiliated Hefei Hospital of Anhui Medical University, Hefei, China

    Deping Xu

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  1. Mingming Zhang
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  2. Tianyu Li
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  4. Yuying Zhang
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  9. Fan He
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  11. Deguang Wang
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Contributions

HZ and DW conceived and designed the experiments. MZ, TL, ZT, YZ, XW and DZ performed the experiments. MZ, HZ, and DW interpreted the data that contributed to the manuscript structure and flow. YF, FH, and MN collected the experimental specimens. MZ, DW, and HZ wrote the manuscript. All authors reviewed and confirmed the manuscript.

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Correspondence to Deguang Wang.

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Zhang, M., Li, T., Tu, Z. et al. Both high glucose and phosphate overload promote senescence-associated calcification of vascular muscle cells. Int Urol Nephrol 54, 2719–2731 (2022). https://doi.org/10.1007/s11255-022-03195-4

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