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Abstract：
Perivascular adipose-derived stem cells (PV-ADSCs) could differentiate into smooth muscle cells (SMCs), participating in vascular remodeling. However, its underlying mechanism is not well explored. Our previous single-cell RNA-sequencing dataset identified a unique expression of matrix Gla protein (MGP) in PV-ADSCs compared with subcutaneous ADSCs. MGP involves in regulating SMC behaviors in vascular calcification and atherosclerosis. In this study, we investigated MGP\'s role in PV-ADSCs differentiation toward SMCs in vitro and in vascular remodeling in vivo. PV-ADSCs were isolated from perivascular regions of mouse aortas. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR), Western blot, and immunofluorescence confirmed higher MGP expression in PV-ADSCs. The MGP secretion increased along PV-ADSCs differentiation toward SMCs in response to transforming growth factor-beta 1 (TGF-β1). Lentivirus knockdown of MGP markedly promoted the bone morphogenetic protein 2 (BMP2) expression and phosphorylation of SMAD1/5/8 in PV-ADSCs, subsequently inhibiting its differentiation toward SMCs. Such inhibition could be partially reversed by further application of BMP2 inhibitors. On the contrary, exogenous MGP inhibited BMP2 expression and SMAD1/5/8 phosphorylation in PV-ADSCs, thereby promoting its differentiation toward SMCs. Transplantation of cultured PV-ADSCs, which was pretreated by MGP knockdown, in mouse femoral artery guide-wire injury model significantly alleviated neointimal hyperplasia. In conclusion, MGP promoted the differentiation of PV-ADSCs toward SMCs through BMP2/SMAD-mediated signaling pathway. This study offers a supplement to the society of perivascular tissues and PV-ADSCs.
摘要：
血管周脂肪干细胞(PV-ADSCs)可以分化为平滑肌细胞(SMCs),参与血管重塑。然而,其潜在机制尚未得到很好的探索。我们之前的单细胞RNA测序数据集鉴定了与皮下ADSC相比，PV-ADSC中基质Gla蛋白（MGP）的独特表达。MGP参与调节SMC在血管钙化和动脉粥样硬化中的行为。在这项研究中,我们研究了MGP在体外PV-ADSCs向SMCs分化和体内血管重塑中的作用。从小鼠主动脉的血管周围区域分离PV-ADSC。定量逆转录-聚合酶链反应（qRT-PCR），蛋白质印迹,免疫荧光证实在PV-ADSCs中有较高的MGP表达。响应于转化生长因子-β1(TGF-β1),MGP分泌随着PV-ADSCs向SMC分化而增加。慢病毒敲除MGP显著促进PV-ADSCs骨形态发生蛋白2(BMP2)的表达和SMAD1/5/8的磷酸化,随后抑制其向SMC的分化。这种抑制可以通过进一步应用BMP2抑制剂而部分逆转。相反,外源性MGP抑制PV-ADSCs中BMP2的表达和SMAD1/5/8的磷酸化,从而促进其向SMC的分化。培养的PV-ADSCs的移植，通过MGP敲除预处理，在小鼠股动脉导丝损伤模型中，可显着减轻新生内膜增生。总之,MGP通过BMP2/SMAD介导的信号通路促进PV-ADSCs向SMCs分化。这项研究为血管周组织和PV-ADSCs的社会提供了补充。
