Vascular Function Laboratory, Human Aging Research Institute and School of Life Science, Nanchang university, and Jiangxi Key Laboratory of Human Aging, Nanchang, China.
Institute of Translational Medicine, Nanchang University, Nanchang, China.
Aging Cell. 2022 Sep;21(9):e13699. doi: 10.1111/acel.13699. Epub 2022 Aug 25.
Hypertension is common in elderly population. We designed to search comprehensively for genes that are chronologically shifted in their expressions and to define their contributions to vascular aging and hypertension. RNA sequencing was conducted to search for senescence-shifted transcripts in human umbilical vein endothelial cells (HUVECs). Small interfering RNA (siRNA), small-molecule drugs, CRISPR/Cas9 techniques, and imaging were used to determine genes' function and contributions to age-related phenotypes of the endothelial cell and blood vessel. Of 25 genes enriched in the term of "regulation of blood pressure," NPRA was changed most significantly. The decreased NPRA expression was replicated in aortas of aged mice. The knockdown of NPRA promoted HUVEC senescence and it decreased expressions of protein kinase cGMP-dependent 1 (PKG), sirtuin 1 (SIRT1), and endothelial nitric oxide synthase (eNOS). Suppression of NPRA also decreased the phosphorylation of AMP-activated protein kinase (AMPK) as well as the ratio of oxidized nicotinamide adenine dinucleotide (NAD )/reduced nicotinamide adenine dinucleotide (NADH) but increased the production of reactive oxygen species (ROS). 8-Br-cGMP (analog of cGMP), or AICAR (AMPK activator), counteracted the observed changes in HUVECs. The Npr1 mice presented an elevated systolic blood pressure and their vessels became insensitive to endothelial-dependent vasodilators. Further, vessels from Npr1 mice increased Cdkn1a but decreased eNos expressions. These phenotypes were rescued by intravenously administrated 8-Br-cGMP and viral overexpression of human PKG, respectively. In conclusion, we demonstrate NPRA/PKG/AMPK as a novel and critical signaling axis in the modulation of endothelial cell senescence, vascular aging, and hypertension.
高血压在老年人群中很常见。我们旨在全面寻找表达时程改变的基因，并定义它们对血管衰老和高血压的贡献。我们对人脐静脉内皮细胞（HUVEC）中的衰老相关转录本进行了 RNA 测序。采用小干扰 RNA（siRNA）、小分子药物、CRISPR/Cas9 技术和成像技术来确定基因的功能及其对内皮细胞和血管与年龄相关表型的贡献。在“血压调节”这一术语中富集的 25 个基因中，NPRA 的变化最为显著。在老年小鼠的主动脉中也复制到 NPRA 表达降低的现象。NPRA 的敲低促进了 HUVEC 的衰老，降低了蛋白激酶 cGMP 依赖性 1（PKG）、SIRT1 和内皮型一氧化氮合酶（eNOS）的表达。NPRA 的抑制还降低了 AMP 激活蛋白激酶（AMPK）的磷酸化以及氧化型烟酰胺腺嘌呤二核苷酸（NAD）/还原型烟酰胺腺嘌呤二核苷酸（NADH）的比值，但增加了活性氧（ROS）的产生。8-Br-cGMP（cGMP 类似物）或 AICAR（AMPK 激活剂）逆转了 HUVEC 中的观察到的变化。Npr1 小鼠表现出收缩压升高，并且它们的血管对内皮依赖性血管舒张剂变得不敏感。此外，Npr1 小鼠的血管增加了 Cdkn1a 但降低了 eNos 的表达。这些表型分别通过静脉内给予 8-Br-cGMP 和病毒过表达人 PKG 得到挽救。总之，我们证明了 NPRA/PKG/AMPK 作为调节内皮细胞衰老、血管衰老和高血压的一个新的和关键的信号轴。
