Matrine blocks AGEs- induced HCSMCs phenotypic conversion via suppressing Dll4-Notch pathway

Abstract

Vascular smooth muscle cells (VSMCs) contractile- synthetic phenotypic conversion takes responsibility in the atherosclerotic plaque formation by abnormal synthesis, secretion and deposition of extracellular matrix (ECM). Matrine exerts therapeutic effects on both cardiovascular diseases and organ fibrosis. In this study, we investigated matrine's inhibitory effect and mechanisms on AGEs- induced VSMC contractile- synthetic phenotypic conversion. Cultured human coronary smooth muscle cells (HCSMCs) were exposed to AGEs. Matrine at serially diluted concentrations were used to treat the cells. HCSMCs phenotype was identified by immunofluorescent staining of contractile phenotypic markers including mooth muscle myosin heavy chain (MYH11) and smooth muscle α-actin (ACTA2). Sircol collagen assay was used to assess the collagen secretion level. Notch signaling activation was determined by luciferase assay. Western blotting was used to evaluate expression levels of collagen I, collagen VIII, Delta-like (Dll)1, Dll3, Dll4, Jagged1, Jagged2, Notch intracellular domain (NICD)1 and Hes family basic helix-loop-helix (bHLH) transcription factor1 (HES1). Matrine pre-treatment recovered the AGEs- induced contractile- synthetic phenotypic conversion by increasing MYH11 and ACTA2 in HCSMCs. Matrine reduced AGEs- mediated activation of Notch signaling, down-regulated expression levels of NICD1, HES1, collagen I and collagen VIII and collagen secretion contents in HCSMCs. Matrine inhibited expression level of Dll4 without affecting other Notch ligands including Dll1, Dll3, Jagged1 and Jagged2 in HCSMCs exposed to AGEs. These results suggested that AGEs exposure facilitated the contractile- synthetic phenotypic conversion of HCSMCs. Matrine blocked this phenotypic conversion by suppressing Dll4- Notch signaling pathway activation.