Adipose-Derived Stem Cells Transfected with GCP-2 Alleviate Pulmonary Arterial Hypertension in Rats by Modulating Apelin/APJ System and eNOS Signaling

Abstract

Objectives: Pulmonary arterial hypertension (PAH) is a disease characterized by the dysfunction of the lung endothelium and angiogenesis. In this study, we aimed to investigate the effects of granulocyte chemoattractant protein 2 (GCP-2) overexpression on adipose-derived stem cells (ADSCs) and the role of ADSCs overexpressing GCP-2 (ADSC-GCP-2) in PAH. Methods: ADSCs were obtained from the adipose tissues of rats and the expression of CD29 and CD45 surface antigens was detected by flow cytometry. The symptoms and tissue damage of PAH rats induced by monocrotaline were evaluated by observing the weight, heart rate, mean blood pressure (MBP) and hematoxylin-eosin (HE) staining. The mRNA and protein expression of genes were tested by Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot, respectively. Transwell® and tube formation assays were adopted to measure cell migration and tube formation capacity. Finally, the levels of vascular regulatory and inflammatory factors were measured by ELISA (Enzyme-Linked Immunosorbent Assay). Results: The improvement of cell migration ability, angiogenesis ability (upregulated VEGF (Vascular Endothelial Growth Factor)-A, FGF2 and TGF-β1), tube formation ability and decreased proinflammatory factors levels (downregulated IL (Interleukin) and TNF (Tumor Necrosis Factor)-α) in MBP induced by monocrotaline was significantly downregulated in the rats with ADSC-GCP-2 treatment compared with ADSC-vector, accompanied by improvement in pulmonary artery thickening and vascular remodeling (downregulated VEGF, HGF and α-actin). Moreover, ADSC-GCP-2 could also significantly reduce tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-17A, and IL-23 levels and increase IL-10 levels to alleviate inflammation in PAH rats (p < 0.01). In addition, ADSC-GCP-2 activated the apelin/APJ (Angiotensin Receptor-Like 1) and eNOS signaling pathways in PAH rats. Conclusions: ADSC-GCP-2 significantly alleviated PAH progression by regulating the apelin/APJ and eNOS signaling pathways. This study provides a potential therapeutic direction for treating PAH.