α-klotho通过抑制Wnt3a/GSK-3β/mTOR信号通路恢复自噬来改善血管紧张素II诱导的内皮细胞衰老：α-klotho在冠状动脉粥样硬化疾病预后中的潜在机制。

Klotho ameliorates angiotension-II-induced endothelial senescence via restoration of autophagy by inhibiting Wnt3a/GSK-3β/mTOR signaling: A potential mechanism involved in prognostic performance of Klotho in coronary atherosclerotic disease.

作者信息

Mao Qi, Deng Mengyang, Zhao Jianhua, Zhou Denglu, Tong Wuyang, Xu Shangcheng, Zhao Xiaohui

机构信息

Department of Cardiology, Institute of Cardiovascular Research, Xinqiao Hospital of Army Medical University, Chongqing 400037, China.

Center of Laboratory Medicine, Chongqing Prevention and Treatment Center for Occupational Diseases, Chongqing 400060, China; Chongqing Key Laboratory of Prevention and Treatment for Occupational Diseases and Poisoning, Chongqing 400060, China.

出版信息

Mech Ageing Dev. 2023 Apr;211:111789. doi: 10.1016/j.mad.2023.111789. Epub 2023 Feb 9.

DOI:10.1016/j.mad.2023.111789

PMID:36764463

Abstract

OBJECTIVE

We aimed to evaluate the prognostic performance of circulating Klotho in coronary atherosclerotic disease (CAD), and to further explore the effect of Klotho on stress-mediated endothelial senescence and underlying mechanism.

METHODS

A cohort of 295 patients had a 12-month follow-up for major adverse cardiovascular events (MACE). Serum Klotho was detected by enzyme linked immunosorbent assay. Cell viability, SA-β-Gal staining, the expression of P53 and P16 were analyzed for endothelial senescence. Oxidative stress was evaluated by measurement of reactive oxygen species, superoxide dismutase and malondialdehyde. LC3, P62, Wnt3a, GSK-3β and mTOR were analyzed by western blotting. Autophagosome formation was detected by adenovirus transfection.

RESULTS

In epidemiological analysis, low Klotho (≤295.9 pg/ml) was significantly associated with MACE risk (HR=2.266, 95 %CI 1.229-4.176). In experimental analysis, Klotho alleviated endothelial senescence and oxidative stress caused by Ang-II exposure; Klotho restored impaired autophagic flux to ameliorate Ang-II induced endothelial senescence; Ang-II activated Wnt3a/GSK-3β/mTOR signaling to inhibit autophagy, whereas Klotho restored autophagy through blockade of Wnt3a/GSK-3β/mTOR signaling; Klotho ameliorated endothelial senescence by suppressing Wnt3a/GSK-3β/mTOR pathway under Ang-II exposure.

CONCLUSIONS

Prognostic significance of Klotho in CAD is potentially ascribed to its anti-endothelial senescence effect via autophagic flux restoration by inhibiting Wnt3a/ GSK-3β/mTOR signaling.

摘要

目的

我们旨在评估循环中α-klotho蛋白在冠状动脉粥样硬化性心脏病（CAD）中的预后价值，并进一步探讨α-klotho蛋白对压力介导的内皮细胞衰老的影响及其潜在机制。

方法

对295例患者进行为期12个月的主要不良心血管事件（MACE）随访。采用酶联免疫吸附测定法检测血清α-klotho蛋白水平。分析细胞活力、衰老相关β-半乳糖苷酶（SA-β-Gal）染色、P53和P16的表达以评估内皮细胞衰老。通过测量活性氧、超氧化物歧化酶和丙二醛来评估氧化应激。采用蛋白质印迹法分析微管相关蛋白1轻链3（LC3）、p62、Wnt3a、糖原合成酶激酶3β（GSK-3β）和哺乳动物雷帕霉素靶蛋白（mTOR）。通过腺病毒转染检测自噬体形成。

结果

在流行病学分析中，低α-klotho蛋白水平（≤295.9 pg/ml）与MACE风险显著相关（HR=2.266，95%CI 1.229-4.176）。在实验分析中，α-klotho蛋白可减轻血管紧张素II（Ang-II）暴露引起的内皮细胞衰老和氧化应激；α-klotho蛋白恢复受损的自噬流以改善Ang-II诱导的内皮细胞衰老；Ang-II激活Wnt3a/GSK-3β/mTOR信号通路以抑制自噬，而α-klotho蛋白通过阻断Wnt3a/GSK-3β/mTOR信号通路恢复自噬；在Ang-II暴露下，α-klotho蛋白通过抑制Wnt3a/GSK-3β/mTOR信号通路改善内皮细胞衰老。