Molecular and cellular pharmacologyMatrine blocks AGEs- induced HCSMCs phenotypic conversion via suppressing Dll4-Notch pathway
Introduction
Coronary artery heart disease is highly associated with diabetes, which is characterized by formation and progression of atherosclerotic plaques (Harper et al., 2016). Under circumstance of hyperglycemia, set of high-molecular weight protein and fluorescent entities are produced after non-enzymatic glycation reactions, referred as advanced glycation end products (AGEs)(Vlassara and Uribarri, 2014). It was believed that AGEs was an indicator of adverse outcomes of atherosclerosis (Ahmad et al., 2017; Falcone et al., 2013). Previous investigations have proved that AGEs induced pathological changes of atherosclerosis such as proliferation and migration in arterial and vascular smooth muscle cells (VSMCs)(San Martin et al., 2007).
As a component of arterial vessel, dysfunctions of VSMCs are involved in the occurrence and progression of atherosclerosis (Bennett et al., 2016). VSMCs behave a range of phenotypes (Thyberg, 2002). Under normal physiological conditions, the VSMCs are at their quiescent status which is also referred as contractile phenotype (Tang et al., 2017), maintaining blood vessel tone by performing constriction and relaxation (Neuman et al., 2009). This phenotype is marked by specific contractile proteins including smooth muscle myosin heavy chain (MYH11) and smooth muscle α-actin (ACTA2)(Owens et al., 2004). When encountering harmful stimuli, the phenotype conversion takes place (Rensen et al., 2007). VSMCs lose expressions of the contractile proteins and produce excessive extracellular matrix (ECM) proteins, participating in vessel stenosis and atherosclerosis.
Notch receptors are highly conserved, playing critical roles in many cellular biological processes (Yamamoto et al., 2014). Delta-like (Dll1, Dll3 and Dll4) and Serrate (Jagged1, Jagged2) are recognized as the ligands of Notch receptors (Hori et al., 2013). These ligands would result in cleavage of Notch receptors and release of Notch intracellular domain (NICD) in to plasma (Tagami et al., 2008) which further initiates expression of its targeted genes (Gokulan and Halagowder, 2014). Notch signaling pathway was proposed to govern the phenotypic modulation of VSMCs (Lin and Lilly, 2014). Another previous study pointed out that the activation of Notch signaling lead to synthesis of ECM (Dees et al., 2011). Moreover, the blockage of Dll4 by specific antibody attenuated atherosclerosis (Fukuda et al., 2012). These above information raised our interests in investigating the possible involvement of Dll4- Notch signaling in the contractile- synthetic phenotype conversion of VSMCs.
Matrine (C15H24N2O) (Fig. 1A) is one of the bio-active molecules of Sophora alpecuroides L. have been attracting researchers’ attention due to its wide spectrum of biological activities and bio-safety. In our previous studies, we observed matrine attenuated cardiac fibrosis in experimental diabetic cardiomyopathy. A recently published study showed that parathyroid hormone, a Notch signaling pathway activator, impaired matrine's anti-fibrotic activity (Yang et al., 2016). Thus, we hypothesize that matrine participates in VSMC phenotypic conversion by regulating Notch signaling. In the current study, human coronary smooth muscle cells (HCSMCs) were exposed to AGEs. The involvement of Notch signaling pathway in HCSMCs phenotypic conversion was investigated. The anti-fibrotic effect of matrine on AGEs- exposed HCSMCs was also studied. We believe that results from our study would not only add current knowledge of diabetic atherogenesis, but also provide novel theoretical basis for clinical application of matrine.
Section snippets
AGEs- bovine serum albumin (BSA) preparation
The AGEs-BSA was prepared in accordance with previous descriptions. Briefly, BSA (Hyclone) was incubated with 0.1 mmol/l glyceraldehydes (Sigma-Aldrich) in 0.2 mmol/l NaPO4 buffer solution (pH = 7.4) at sterile condition at 37 ℃ for 7 days. Control BSA was simultaneously prepared without incubation with glyceraldehydes.
Cell culture
HCSMCs were provided by PromoCell (Germany) and maintained in Dulbecco's modified Eagle's medium (DMEM, Gibco) supplemented with antibiotic mix (Invitrogen) in a humidified
AGEs incubation facilitated the contractile- synthetic phenotypic conversion of HCSMCs
As demonstrated in Fig. 2A, AGEs incubation significantly reduced the expression levels of molecular markers of contractile phenotype of VSMCs, namely MYH11 and ACTA2 in HCSMCs. Moreover, also shown in Fig. 2B and C, the expression levels of collagen proteins as wells as the collagen secretion content were dramatically increased by AGEs incubation in HCSMCs.
AGEs incubation elevated expression of Notch ligands which activated Notch signaling
The results were demonstrated in Fig. 3. The expression levels of Notch ligands including Dll1, Dll3, Dll4, Jagged1 and Jagged2 were
Discussion
VSMCs are critical for maintaining the integrity and normal function of arteries. VSMCs display a spectrum of different phenotypes under different pathophysiological conditions to meet current needs of the arterial vascular system (Zhang et al., 2016). Due to alterations of lifestyle and diet habits, the mortality and morbidity of diabetes is increasing fast in both developing and developed countries (Echouffo-Tcheugui and Dagogo-Jack, 2012). Uncontrolled and sustained hyperglycemia is the most
Funding
This work received support from the following financial sources:
- 1.
Special Financial Grant from the China Postdoctoral Science Foundation (No. 2017T100760)
- 2.
China Postdoctoral Science Foundation (No. 2016M590956)
- 3.
National Scientific Foundation of China (No. 81600646)
- 4.
Sailing Foundation (No. LHJJ20159029)
References (42)
- et al.
Notch transcriptional control of vascular smooth muscle regulatory gene expression and function
J. Biol. Chem.
(2013) - et al.
Advanced glycation end-products: implications for diabetic and non-diabetic nephropathies
Diabetes Metab.
(2010) - et al.
Vascular calcification in type-2 diabetes and cardiovascular disease: integrative roles for OPG, RANKL and TRAIL
Vasc. Pharmacol.
(2016) - et al.
Notch signaling governs phenotypic modulation of smooth muscle cells
Vasc. Pharmacol.
(2014) - et al.
Matrine attenuates cardiac fibrosis by affecting ATF6 signaling pathway in diabetic cardiomyopathy
Eur. J. Pharmacol.
(2017) - et al.
Type VIII collagen mediates vessel wall remodeling after arterial injury and fibrous cap formation in atherosclerosis
Am. J. Pathol.
(2013) - et al.
Notch ligand endocytosis: mechanistic basis of signaling activity
Semin. Cell Dev. Biol.
(2012) - et al.
The four-and-a-half LIM domain protein 2 regulates vascular smooth muscle phenotype and vascular tone
J. Biol. Chem.
(2009) - et al.
Ligand-independent mechanisms of notch activity
Trends Cell Biol.
(2015) - et al.
Nox1-based NADPH oxidase-derived superoxide is required for VSMC activation by advanced glycation end-products
Free Radic. Biol. Med.
(2007)
Advanced glycation endproducts in diabetes and diabetic complications
Endocrinol. Metab. Clin. N. Am.
Effect of matrine on transforming growth factor beta1 and hepatocyte growth factor in rat liver fibrosis model
Asian Pac. J. Trop. Med.
Akt1 isoform modulates phenotypic conversion of vascular smooth muscle cells
Biochim. Biophys. Acta
A glycation angle to look into the diabetic vasculopathy: cause and cure
Curr. Vasc. Pharmacol.
Vascular smooth muscle cells in atherosclerosis
Circ. Res.
HES1, a target of Notch signaling, is elevated in canine osteosarcoma, but reduced in the most aggressive tumors
BMC Vet. Res.
Notch signalling regulates fibroblast activation and collagen release in systemic sclerosis
Ann. Rheum. Dis.
Preventing diabetes mellitus in developing countries
Nat. Rev. Endocrinol.
Plasma levels of soluble receptor for advanced glycation end products and coronary atherosclerosis: possible correlation with clinical presentation
Dis. Markers
Notch ligand delta-like 4 blockade attenuates atherosclerosis and metabolic disorders
Proc. Natl. Acad. Sci. USA
Expression pattern of Notch intracellular domain (NICD) and Hes-1 in preneoplastic and neoplastic human oral squamous epithelium: their correlation with c-Myc, clinicopathological factors and prognosis in oral cancer
Med. Oncol.
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