Julius-Bernstein-Institute of Physiology, Martin Luther University Halle-Wittenberg, Halle, Germany.
Acta Physiol (Oxf). 2021 Nov;233(3):e13715. doi: 10.1111/apha.13715. Epub 2021 Jul 16.
This study investigates the role of calcineurin for angiotensin II (AngII)-induced vascular remodelling with the help of a mouse model lacking the catalytic beta subunit of calcineurin (PPP3CB KO).
Wildtype (WT) and PPP3CB KO mice were treated for 4 weeks with AngII followed by assessment of blood pressure, histological evaluation of aortas and mRNA analysis of aortic genes PPP3CB-dependently regulated by AngII. Primary murine vascular smooth muscle cells (VSMCs) were used for qPCR, ELISA and Western Blot experiments as well as wound healing and cell proliferation assays.
Upon AngII treatment, PPP3CB KO mice showed less aortic media thickening, lumen dilation and systolic blood pressure compared to WT mice. Next-generation sequencing data of aortic tissue indicated an increase in extracellular matrix components (EMCs), cell migration and cell proliferation. A PPP3CB-dependent increase in EMC was confirmed by qPCR in aorta and VSMCs. PPP3CB-dependent stimulation of VSMC migration could be verified by wound healing assays but markers of enhanced cell proliferation were only detectable in aortic tissue of WT mice but not in isolated WT or KO VSMCs. We could demonstrate in VSMCs with pharmacological inhibitors that PPP3CB leads to enhanced heparin-binding EGF-like growth factor (HB-EGF) secretion, epidermal growth factor receptor (EGFR) activation and consecutive stimulation of transforming growth factor β(TGFβ) and connective tissue growth factor (CTGF) signalling that enhances collagen expression.
AngII-induced vascular remodelling involves PPP3CB, which leads to enhanced EMC production, VSMC migration and sustained increase in systolic blood pressure via HBEGF/EGFR-TGFβ-CTGF signalling.
本研究借助缺乏钙调神经磷酸酶催化β亚基（PPP3CB KO）的小鼠模型，探讨钙调神经磷酸酶在血管紧张素 II（AngII）诱导的血管重构中的作用。
野生型（WT）和 PPP3CB KO 小鼠接受 AngII 治疗 4 周，然后评估血压、主动脉组织学评估和 AngII 依赖调节的主动脉基因的 mRNA 分析。原代小鼠血管平滑肌细胞（VSMCs）用于 qPCR、ELISA 和 Western Blot 实验以及伤口愈合和细胞增殖实验。
在 AngII 处理后，PPP3CB KO 小鼠的主动脉中层增厚、管腔扩张和收缩压均低于 WT 小鼠。主动脉组织的下一代测序数据表明细胞外基质成分（EMCs）、细胞迁移和细胞增殖增加。qPCR 在主动脉和 VSMCs 中证实了 PPP3CB 依赖性 EMC 增加。伤口愈合实验证实了 PPP3CB 依赖性 VSMC 迁移刺激，而增强的细胞增殖标志物仅在 WT 小鼠的主动脉组织中检测到，而在分离的 WT 或 KO VSMCs 中未检测到。我们在 VSMCs 中用药理学抑制剂证明 PPP3CB 导致肝素结合表皮生长因子样生长因子（HB-EGF）分泌、表皮生长因子受体（EGFR）激活和转化生长因子 β（TGFβ）和结缔组织生长因子（CTGF）信号的连续刺激增强胶原表达。
AngII 诱导的血管重构涉及 PPP3CB，它通过 HBEGF/EGFR-TGFβ-CTGF 信号通路导致 EMC 产生增加、VSMC 迁移和收缩压持续升高。
