MGP regulates perivascular adipose-derived stem cells differentiation towards smooth muscle cells via BMP2/SMAD pathway enhancing neointimal formation
MGP regulates perivascular adipose-derived stem cells differentiation towards smooth muscle cells via BMP2/SMAD pathway enhancing neointimal formation
复制标题
MGP 通过 BMP2/SMAD 通路调节血管周围脂肪干细胞向平滑肌细胞分化，增强新内膜形成
作者：
Ni Hui;Liu Chang;Chen Yuwen;Lu Yunrui;Ji Yongli;Xiang Meixiang;Xie Yao
血管周围脂肪干细胞（PV-ADSCs）可分化为平滑肌细胞（SMCs），参与血管重塑。然而，其潜在的机制并没有得到很好的探讨。我们先前的单细胞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参与调节血管钙化和动脉粥样硬化中SMCs的行为。在本研究中，我们研究了MGP在体外PV-ADSCs向SMC分化中的作用，以及在体内血管重塑中的作用。PV-ADSC分离自小鼠睾丸的血管周围区域。QRT-PCR、Western印迹和免疫荧光证实PV-ADSC中MGP表达较高。TGF-β1诱导PV-ADSC向SMCs分化时，MGP分泌沿着增加。慢病毒基因敲减MGP可显著促进PV-ADSCs中BMP 2的表达和SMAD 1/5/8的磷酸化，从而抑制PV-ADSCs向SMCs的分化。这种抑制可以通过进一步应用BMP 2抑制剂部分逆转。相反，外源性MGP抑制PV-ADSC中BMP 2的表达和SMAD 1/5/8磷酸化，从而促进其向SMCs分化。经MGP基因敲减处理的PV-ADSCs移植于小鼠股动脉导丝损伤模型，可明显减轻新生内膜增生。结论：MGP通过BMP 2/SMAD信号通路促进PV-ADSCs向平滑肌细胞分化。本研究为PV-ADSCs与血管周围组织的研究提供了补充。
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 to subcutaneous ADSCs. MGP involves in regulating SMCs behaviors in vascular calcification and atherosclerosis. In present study, we investigated MGP’s role in PV-ADSCs differentiation towards SMCs in vitro, and in vascular remodeling in vivo. PV-ADSCs were isolated from perivascular regions of mouse aortas. QRT-PCR, Western blot, and immunofluorescence confirmed higher MGP expression in PV-ADSCs. The MGP secretion increased along PV-ADSCs differentiation towards SMCs in response to TGF-β1. Lentivirus knockdown of MGP markedly promoted the BMP2 expression and phosphorylation of SMAD1/5/8 in PV-ADSCs, subsequently inhibiting its differentiation towards 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 towards 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 towards SMCs through BMP2/SMAD-mediated signaling pathway. The present study offered a supplement to the society of perivascular tissues and PV-ADSCs.