Dissecting Molecular Mechanisms Underlying Pulmonary Vascular Smooth Muscle Cell Dedifferentiation in Pulmonary Hypertension: Role of Mutated Caveolin-1 (Cav1F92A)-Bone Marrow Mesenchymal Stem Cells

Wancheng Yu; Haiying Chen; Hongli Yang; Jie Ding; Peng Xia; Xu Mei; Lei Wang; Shuangfeng Chen; Chengwei Zou; Le-Xin Wang

doi:10.1016/j.hlc.2018.08.002

Dissecting Molecular Mechanisms Underlying Pulmonary Vascular Smooth Muscle Cell Dedifferentiation in Pulmonary Hypertension: Role of Mutated Caveolin-1 (Cav1^F92A)-Bone Marrow Mesenchymal Stem Cells

Heart Lung Circ. 2019 Oct;28(10):1587-1597. doi: 10.1016/j.hlc.2018.08.002. Epub 2018 Sep 7.

Authors

Wancheng Yu¹, Haiying Chen², Hongli Yang², Jie Ding², Peng Xia³, Xu Mei⁴, Lei Wang³, Shuangfeng Chen², Chengwei Zou⁵, Le-Xin Wang⁶

Affiliations

¹ Department of Cardiac Surgery, Provincial Hospital Affiliated to Shandong University, Shandong 250021, China.
² Central laboratory of Liaocheng People's Hospital, Liaocheng, Shandong, 252000, China.
³ Department of Cardiology, Liaocheng People's Hospital and Affiliated Liaocheng People's Hospital of Shandong University, Liaocheng, Shandong, 252000, China.
⁴ Department of Geriatrics, Shandong University Qilu Hospital, Shandong, China.
⁵ Department of Cardiac Surgery, Provincial Hospital Affiliated to Shandong University, Shandong 250021, China. Electronic address: zouchengwei@sdu.edu.cn.
⁶ Department of Cardiology, Liaocheng People's Hospital and Affiliated Liaocheng People's Hospital of Shandong University, Liaocheng, Shandong, 252000, China; School of Biomedical Sciences, Charles Sturt University, Wagga Wagga, NSW, Australia. Electronic address: lwang@csu.edu.au.

PMID: 30262154
DOI: 10.1016/j.hlc.2018.08.002

Abstract

Background: Pulmonary arterial hypertension (PAH) is characterised by remodelling in vascular smooth muscles, and switching from contractile (differentiated) to synthetic (dedifferentiated) phenotype. This study aimed to investigate the effect of a mutated caveolin-1 (Cav1^F92A) gene from bone marrow mesenchymal stem cells (rBMSCs) on phenotypic switching in the smooth muscle cells during PAH.

Methods: Human pulmonary smooth muscle cells (HPASMCs) were treated with monocrotaline (MCT,1μM), and co-cultured with Cav1^F92A gene modified rBMSCs (rBMSCs/Cav1^F92A). The nitric oxide (NO) production, cell adhesion, cell viability and inflammatory cytokines expression in rBMSCs was measured to evaluate the survival rate of rBMSCs and the changes of inflammatory cytokines. The concentration of NO/cGMP (nitric oxide/Guanosine-3',5'-cyclic monophosphate), the tumour necrosis factor-alpha (TNF-α), transforming growth factor-beta1 (TGF-β1) mRNA, the expression of contractile smooth muscle cells (SMCs) phenotype markers (thrombospondin-1 and Matrix Gla protein, MGP), the synthetic SMCs phenotype markers (H-caldesmon and smooth muscle gene SM22 alpha, SM22α), cell migration and the morphological changes in rBMSCs/Cav1^F92A co-cultured HPASMCs were investigated.

Results: Cav1^F92A increased NO concentration, cell adhesion, cell viability, anti-inflammatory cytokines interleukin-4 (IL-4), and interleukin-10 (IL-10), but decreased the inflammatory cytokines interleukin-1α (IL-1α), interferon-γ (INF-γ) and TNF-α expression in rBMSCs. rBMSCs/Cav1^F92A activated the NO/cGMP, down-regulated TNF-α, TGF-β1, thrombospondin-1 and MGP expression, up-regulated SM22α and H-caldesmon expression, restored cell morphology, and inhibited cell migration in MCT treated HPASMCs.

Conclusions: rBMSCs/Cav1^F92A inhibits switching from contractile to synthetic phenotype in HPASMCs. It also inhibits migration and promotes morphological restoration of these cells. rBMSCs/Cav1^F92A may be used as a therapeutic modality for PAH.

Keywords: Human pulmonary artery smooth muscle cells; Mutation caveolin-1(Cav1(F92A)); Nitric oxide; Phenotypic switching; Rat bone marrow mesenchymal stem cells.

MeSH terms

Caveolin 1 / genetics*
Caveolin 1 / metabolism
Cell Dedifferentiation
Cell Movement
Cell Proliferation
Cells, Cultured
DNA / genetics*
DNA Mutational Analysis
Humans
Hypertension, Pulmonary / genetics*
Hypertension, Pulmonary / metabolism
Hypertension, Pulmonary / pathology
Mesenchymal Stem Cells / cytology
Mesenchymal Stem Cells / metabolism*
Muscle, Smooth, Vascular / metabolism*
Muscle, Smooth, Vascular / pathology
Mutation*
Pulmonary Artery / metabolism*
Pulmonary Artery / pathology

Substances

CAV1 protein, human
Caveolin 1
DNA