环状 RNA 相互作用蛋白激酶 3 通过与 FUS 相互作用介导沉默信息调节因子 1/过氧化物酶体增殖物激活受体 γ 共激活因子 1α/线粒体融合蛋白 2 通路缓解血管钙化。
CircHIPK3 relieves vascular calcification via mediating SIRT1/PGC-1α/MFN2 pathway by interacting with FUS.
机构信息
Department of Ultrasound Medicine, Hainan General Hospital, Hainan Affiliated Hospital of Hainan Medical University, Haikou, 570311, Hainan Province, China.
Department of Vascular Surgery, Hainan General Hospital, Hainan Affiliated Hospital of Hainan Medical University, No.19, Xiuhua Road, Haikou, 570311, Hainan Province, China.
出版信息
BMC Cardiovasc Disord. 2023 Nov 27;23(1):583. doi: 10.1186/s12872-023-03602-3.
BACKGROUND
Circular RNAs (circRNAs) have been reported to regulate the biological processes of human diseases. CircHIPK3 has been implicated in vascular calcification, but the downstream regulatory mechanisms remain unclear. Our study aimed to understand the regulatory function of circHIPK3 in vascular calcification.
METHODS
CircHIPK3 expression in atherosclerosis (AS) serum samples and vascular smooth muscle cells (VSMCs) calcification model was assessed by quantitative real-time polymerase chain reaction (qRT-PCR). The binding relationships between fused in sarcoma (FUS) and circHIPK3 or sirtuin 1 (SIRT1) were verified by RNA immunoprecipitation (RIP) assay and RNA pull-down assays. Alkaline phosphatase (ALP) activity and alizarin red staining assays were performed to evaluate the biological effect of β-glycerophosphate (β-GP) and circHIPK3 on calcium deposition. qRT-PCR and western blot assays were used to examine the effect of β-GP, circHIPK3, SIRT1, mitofusin 2 (MFN2), and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) on VSMCs calcification and the expression of calcification-related proteins.
RESULTS
In AS serum samples and VSMCs calcification model, the expression of circHIPK3 was significantly reduced. CircHIPK3 overexpression inhibited ALP activity and calcium deposition in β-GP-induced VSMCs. Moreover, circHIPK3 could recruit FUS to further stabilize SIRT1 mRNA. CircHIPK3 promoted MFN2 expression to alleviate VSMCs calcification via activating SIRT1/PGC-1α signaling.
CONCLUSION
The positive regulation of circHIPK3/FUS/SIRT1/PGC-1α/MFN2 signaling pathway contributed to the alleviate VSMCs calcification, revealing a novel regulatory axis for vascular calcification.
背景
环状 RNA（circRNAs）已被报道可调节人类疾病的生物学过程。CircHIPK3 已被牵涉到血管钙化，但下游的调控机制尚不清楚。我们的研究旨在了解 circHIPK3 在血管钙化中的调控功能。
方法
通过定量实时聚合酶链反应（qRT-PCR）评估动脉粥样硬化（AS）血清样本和血管平滑肌细胞（VSMCs）钙化模型中的 circHIPK3 表达。通过 RNA 免疫沉淀（RIP）测定和 RNA 下拉测定验证融合肉瘤（FUS）与 circHIPK3 或 Sirtuin 1（SIRT1）之间的结合关系。碱性磷酸酶（ALP）活性和茜素红染色测定用于评估β-甘油磷酸（β-GP）和 circHIPK3 对钙沉积的生物学效应。qRT-PCR 和 Western blot 测定用于研究β-GP、circHIPK3、SIRT1、线粒体融合蛋白 2（MFN2）和过氧化物酶体增殖物激活受体γ共激活因子 1-α（PGC-1α）对 VSMCs 钙化和钙化相关蛋白表达的影响。
结果
在 AS 血清样本和 VSMCs 钙化模型中，circHIPK3 的表达显著降低。circHIPK3 过表达抑制了β-GP 诱导的 VSMCs 中的 ALP 活性和钙沉积。此外，circHIPK3 可以募集 FUS 以进一步稳定 SIRT1 mRNA。circHIPK3 通过激活 SIRT1/PGC-1α 信号通路促进 MFN2 表达，从而减轻 VSMCs 钙化。
结论
circHIPK3/FUS/SIRT1/PGC-1α/MFN2 信号通路的正调控有助于减轻 VSMCs 钙化，为血管钙化提供了一个新的调控轴。
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