The ameliorative effect of terpinen-4-ol on ER stress-induced vascular calcification depends on SIRT1-mediated regulation of PERK acetylation

Abstract

BackgroundEndoplasmic reticulum (ER) stress-mediated phenotypic switching of vascular smooth muscle cells (VSMCs) is key to vascular calci cation (VC) in patients with chronic kidney disease (CKD). Terpinen-4-ol exerts protective effect against cardiovascular disease, but its role and speci c mechanism in VC remain unclear. We explored whether terpinen-4-ol alleviates ER stress-mediated VC through sirtuin 1 (sirt1) and elucidated its mechanism to provide evidence for its application in the clinical prevention and treatment of VC. MethodsIn this study, CKD-related VC animal model and β-glycerophosphate (β-GP)-induced VSMCs calci cation model were established. We investigated the part of terpinen-4-ol in ER stress-induced VC in vitro and in vivo. However, in order to clarify whether terpinen-4-ol inhibits the molecular mechanism of ERs-induced VC through sirt1, we further veri ed the above signal transduction by knocking down sirt1 in vitro and in vivo. ResultsTerpinen-4-ol inhibited calcium deposition, phenotypic switching, and ER stress of VSMCs in vitro and in vivo. Furthermore, pre-incubation with terpinen-4-ol or a sirt1 agonist and transfection with lentivirus overexpressing sirt1 decreased β-GP-induced calcium salt deposition, increased sirt1 protein level, and inhibited PERK-eIF2α-ATF4 pathway activation in VSMCs, thus, alleviating VC. The opposite results were obtained in sirt1-knockdown models. Moreover, sirt1 physically interacted with and deacetylated PERK.Mass spectrometry analysis identi ed lysine K889 as the acetylation site of sirt1, which regulates PERK.Finally, inhibition of sirt1 reduced the effect of terpinen-4-ol on the deacetylation of PERK in vitro and in vivo and weakened the inhibitory effect of terpinen-4-ol against ER stress-mediated VC. ConclusionsTerpinen-4-ol inhibits the post-transcriptional modi cation of PERK at the lysine K889 acetylation site by upregulating sirt1 expression level, thereby ameliorating VC by regulating ER stress. This provides evidence of the molecular mechanism of terpinen-4-ol, which supports its development as a promising therapeutic agent for CKD-VC.

Melatonin promotes sirtuin 1 expression and inhibits IRE1α–XBP1S–CHOP to reduce endoplasmic reticulum stress–mediated apoptosis in chondrocytes

“…It was recently shown that Sirt1 inhibition induces hyperacetylation and phosphorylation of eIF2α and PERK to regulate PERK-ATF4 signaling of ER stress ( Prola et al, 2017 ; Kang et al, 2018 ). Additionally, SIRT1 deacetylates PERK physically through physical interactions ( Zhang et al, 2021 ). It has been shown that SIRT1 deacetylates XBP1s and inhibits the transcriptional activity of XBP1s to regulate UPR signaling ( Wang et al, 2011 ).…”

CircZXDC Promotes Vascular Smooth Muscle Cell Transdifferentiation via Regulating miRNA-125a-3p/ABCC6 in Moyamoya Disease

“…Despite the fact that its structural features have been largely characterized, knowledge with respect to its functions was limited to proton pumping, and not much is known regarding its function in VSMCs [ 55 , 56 ]. Recently, several studies have reported that ABCC6 might be associated with endoplasmic reticulum stress (ERS) [ 27 , 28 ], and ERS was considered a key mediator for VSMC transdifferentiation towards the synthetic state [ 57 , 58 , 59 ]. Interestingly, previous studies have revealed that RNF213 suppression affects ERS, suggesting an intrinsic connection between RNF213 and ERS [ 60 ].…”

Possible effects of fibroblast growth factor 21 on vascular calcification via suppressing activating transcription factor 4 mediated apoptosis and osteogenic transformation in rats

“…25 Among the ERS pathways, the ATF4 pathway is most closely related to the development of VC. 26 ATF4, namely cAMP responseelement binding protein 2 (CREB2), is a common basic/leucine zipper domain transcription factor, which is also in the 5' regulatory region of the human FGF21 gene. 12 When ERS was initiated, ATF4 was activated by one of ERS transducer molecules, PERK, and it can directly up-regulate its downstream molecule, CHOP, to induce osteoblast apoptosis.…”

“…Whereas, the initiation of ERS‐mediated apoptotic process may trigger by multiple signaling pathways 25 . Among the ERS pathways, the ATF4 pathway is most closely related to the development of VC 26 . ATF4, namely cAMP response‐element binding protein 2 (CREB2), is a common basic/leucine zipper domain transcription factor, which is also in the 5' regulatory region of the human FGF21 gene 12 .…”