SIRT1 and Connexin40 Mediate the Normal Shear Stress-Induced Inhibition of the Proliferation of Endothelial Cells Co-Cultured with Vascular Smooth Muscle Cells

Yao, Qing; Qi, Ying‐Xin; Zhang, Ping; Cheng, Baohua; Yan, Zhiqiang; Jiang, Zong‐Lai

doi:10.1159/000343376

Cited by 19 publications

(15 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It should be noted that in the present study, we found that shear stress of 5 dyn/cm 2 decreased the level of Cx40 in HCAECs co-cultured with HCSMCs, while shear stress of 12 dyn/cm 2 had no effect, which is not consistent with the findings in the previous study of HUVECs [38]. This discrepancy may be attributed to the difference in the cell lines used, indirectly supported by the difference of HCAECs and HUVECs in the wound closure under low laminar shear stress (3 dyn/cm 2 ) [39].…”

Section: Discussioncontrasting

confidence: 99%

“…In case of Cx40, a previous study showed that its endothelial protein expression was elevated markedly by shear stress from 6-10 dyn/cm 2 , while higher shear stress did not affect the expression of Cx40 in HUVECs cultured alone compared to static conditions [20]. However, an earlier study showed that shear stress of 15 dyn/cm 2 downregulated the level of Cx40 expression in rat aortic HUVECs co-cultured with HUVSMCs compared to static conditions [38]. These discrepancies might be explained by different cellular culture models used in the studies and suggested that SMCs might affect the mechanical response of ECs to shear stress.…”

Section: Discussionmentioning

confidence: 93%

See 1 more Smart Citation

Role of Myoendothelial Gap Junctions in the Regulation of Human Coronary Artery Smooth Muscle Cell Differentiation by Laminar Shear Stress

Zhang

Chen²,

Zhang³

et al. 2016

Cell Physiol Biochem

View full text Add to dashboard Cite

Background/Aims: Smooth muscle cells may dedifferentiate into the synthetic phenotype and promote atherosclerosis. Here, we explored the role of myoendothelial gap junctions in phenotypic switching of human coronary artery smooth muscle cells (HCASMCs) co-cultured with human coronary artery endothelial cells (HCAECs) exposed to shear stress. Methods: HCASMCs and HCAECs were seeded on opposite sides of Transwell inserts, and HCAECs were exposed to laminar shear stress of 12 dyn/cm² or 5 dyn/cm². The myoendothelial gap junctions were evaluated by using a multi-photon microscope. Results: In co-culture with HCAECs, HCASMCs exhibited a contractile phenotype, and maintained the expression of differentiation markers MHC and H1-calponin. HCASMCs and HCAECs formed functional intercellular junctions, as evidenced by colocalization of connexin(Cx)40 and Cx43 on cellular projections inside the Transwell membrane and biocytin transfer from HCAECs to HCASMCs. Cx40 siRNA and 18-α-GA attenuated protein expression of MHC and H1-calponin in HCASMCs. Shear stress of 5 dyn/cm² increased Cx43 and decreased Cx40 expression in HCAECs, and partly inhibited biocytin transfer from HCAECs to HCASMCs, which could be completely blocked by Cx43 siRNA or restored by Cx40 DNA transfected into HCAECs. The exposure of HCAECs to shear stress of 5 dyn/cm² promoted HCASMC phenotypic switching, manifested by morphological changes, decrease in MHC and H1-calponin expression, and increase in platelet-derived growth factor (PDGF)-BB release, which was partly rescued by Cx43 siRNA or Cx40 DNA or PDGF receptor signaling inhibitor. Conclusions: The exposure of HCAECs to shear stress of 5 dyn/cm² caused the dysfunction of Cx40/Cx43 heterotypic myoendothelial gap junctions, which may be replaced by homotypic Cx43/Cx43 channels, and induced HCASMC transition to the synthetic phenotype associated with the activation of PDGF receptor signaling, which may contribute to shear stress-associated arteriosclerosis.

show abstract

Section: Discussioncontrasting

confidence: 99%

Section: Discussionmentioning

confidence: 93%

Role of Myoendothelial Gap Junctions in the Regulation of Human Coronary Artery Smooth Muscle Cell Differentiation by Laminar Shear Stress

Zhang

Chen²,

Zhang³

et al. 2016

Cell Physiol Biochem

View full text Add to dashboard Cite

show abstract

“…SIRT1 has been proved to play critical roles in vascular homeostasis and remodeling [22,39]. Here, using SIRT1 activator-resveratrol, SIRT1 overexpression plasmid, specific siRNA transfection, and knockout mice, we revealed the protective role of SIRT1 in maintaining the normal contractile differentiation of VSMCs.…”

Section: Discussionmentioning

confidence: 90%

“…To further study the role of SIRT1, FOXO3a and FOXO4 in response to mechanical stretch, resveratrol, an activator of SIRT1 [22] was used to stimulate SIRT1 in VSMCs under static conditions. The results showed that VSMCs treated with resveratrol (1 μM) did not have a significant effect on the expression of SIRT1, whereas 10 μM, 25 μM, 50 μM and 100 μM Resveratrol all significantly increased the expression of SIRT1 (Fig.…”

Section: Resveratrol Modulated Expression Of Foxo3a Foxo4 and Contramentioning

confidence: 99%

SIRT1 and FOXO Mediate Contractile Differentiation of Vascular Smooth Muscle Cells under Cyclic Stretch

Huang

Yan

Zhao

et al. 2015

Cell Physiol Biochem

Self Cite

View full text Add to dashboard Cite

Background/Aims: Physiological mechanical stretch in vivo helps to maintain the quiescent contractile differentiation of vascular smooth muscle cells (VSMCs), but the underlying mechanisms are still unclear. Here, we investigated the effects of SIRT1 in VSMC differentiation in response to mechanical cyclic stretch. Methods and Results: Rat VSMCs were subjected to 10%-1.25Hz-cyclic stretch in vitro using a FX-4000T system. The data indicated that the expression of contractile markers, including α-actin, calponin and SM22α, was significantly enhanced in VSMCs that were subjected to cyclic stretch compared to the static controls. The expression of SIRT1 and FOXO3a was increased by the stretch, but the expression of FOXO4 was decreased. Decreasing SIRT1 by siRNA transfection attenuated the stretch-induced expression of contractile VSMC markers and FOXO3a. Furthermore, increasing SIRT1 by either treatment with activator resveratrol or transfection with a plasmid to induce overexpression increased the expression of FOXO3a and contractile markers, and decreased the expression of FOXO4 in VSMCs. Similar trends were observed in VSMCs of SIRT1 (+/-) knockout mice. The overexpression of FOXO3a promoted the expression of contractile markers in VSMCs, while the overexpression of FOXO4 demonstrated the opposite effect. Conclusion: Our results indicated that physiological cyclic stretch promotes the contractile differentiation of VSMCs via the SIRT1/FOXO pathways and thus contributes to maintaining vascular homeostasis.

show abstract

“…In mice, neointima formation is accompanied by a progressive down-regulation of SIRT1 [19]. In cultured endothelial cells, laminar flow increases whereas oscillating flow decreases the expression and activity of SIRT1 [20, 21]. Despite this information, it is unclear whether or not endothelial SIRT1 modulates arterial remodeling, and if so, by what mechanism(s).…”

Section: Introductionmentioning

confidence: 99%

Endothelial SIRT1 prevents adverse arterial remodeling by facilitating HERC2-mediated degradation of acetylated LKB1

et al. 2016

View full text Add to dashboard Cite

Aims-SIRT1 exerts potent activity against cellular senescence and vascular ageing. By decreasing LKB1 protein levels, it promotes the survival and regeneration of endothelial cells. The present study aims to investigate the molecular mechanisms underlying SIRT1-mediated LKB1 degradation for the prevention of vascular ageing.Methods and Results-Co-immunoprecipitation assay demonstrated that SIRT1, via its amino-terminus, binds to the DOC domain of HERC2 [HECT and RLD domain containing E3 ubiquitin protein ligase 2], which then ubiquitinates LKB1 in the nuclear compartment of endothelial cells. Site-directed mutagenesis revealed that acetylation at lysine (K) 64 of LKB1 triggers the formation of SIRT1/HERC2/LKB1 protein complex and subsequent proteasomal degradation. In vitro cellular studies suggested that accumulation of acetylated LKB1 in the nucleus leads to endothelial activation, in turn stimulating the proliferation of vascular smooth muscle cells and the production of extracellular matrix proteins. Chromatin immunoprecipitation quantitative PCR confirmed that acetylated LKB1 interacts with and activates TGFβ1 promoter, which is inhibited by SIRT1. Knocking down either SIRT1 or HERC2 results in an increased association of LKB1 with the positive regulatory elements of TGFβ1 promoter. In mice without endothelial nitric oxide synthase, selective overexpression of human SIRT1 in endothelium prevents hypertension and age-related adverse arterial remodeling. Lentiviral-mediated knockdown of HERC2 abolishes the beneficial effects of endothelial SIRT1 on both arterial remodeling and arterial blood pressure control.Conclusion-By downregulating acetylated LKB1 protein via HERC2, SIRT1 fine-tunes the crosstalk between endothelial and vascular smooth muscle cells to prevent adverse arterial remodeling and maintain vascular homeostasis.

show abstract

SIRT1 and Connexin40 Mediate the Normal Shear Stress-Induced Inhibition of the Proliferation of Endothelial Cells Co-Cultured with Vascular Smooth Muscle Cells

Cited by 19 publications

References 42 publications

Role of Myoendothelial Gap Junctions in the Regulation of Human Coronary Artery Smooth Muscle Cell Differentiation by Laminar Shear Stress

Role of Myoendothelial Gap Junctions in the Regulation of Human Coronary Artery Smooth Muscle Cell Differentiation by Laminar Shear Stress

SIRT1 and FOXO Mediate Contractile Differentiation of Vascular Smooth Muscle Cells under Cyclic Stretch

Endothelial SIRT1 prevents adverse arterial remodeling by facilitating HERC2-mediated degradation of acetylated LKB1

Contact Info

Product

Resources

About