N-acetylneuraminic acid modulates SQSTM1/p62 sialyation-mediated ubiquitination degradation contributing to vascular endothelium dysfunction in experimental atherosclerosis mice

作者全名："Chen, Le; Qiu, Hongmei; Chen, Qingqiu; Xiang, Peng; Lei, Jin; Zhang, Jun; Lu, Yining; Wang, Xianmin; Wu, Shengde; Yu, Chao; Ma, Limei"

作者地址："[Chen, Le; Qiu, Hongmei; Chen, Qingqiu; Xiang, Peng; Lu, Yining; Wang, Xianmin; Yu, Chao; Ma, Limei] Chongqing Med Univ, Coll Pharm, Chongqing 400010, Peoples R China; [Chen, Le; Qiu, Hongmei; Chen, Qingqiu; Xiang, Peng; Zhang, Jun; Lu, Yining; Wang, Xianmin; Yu, Chao; Ma, Limei] Chongqing Key Lab Pharmaceut Metab Res, Chongqing, Peoples R China; [Lei, Jin] Northwest Univ, Xian 1 Hosp, Affiliated Hosp 1, Xian, Peoples R China; [Wu, Shengde] Chongqing Med Univ, Dept Urol, Childrens Hosp, Chongqing, Peoples R China"

通信作者："Yu, C; Ma, LM (通讯作者)，Chongqing Med Univ, Coll Pharm, Chongqing 400010, Peoples R China."

来源：IUBMB LIFE

ESI学科分类：BIOLOGY & BIOCHEMISTRY

WOS号：WOS:001082155500001

JCR分区：Q2

影响因子：3.7

年份：2023

卷号：　

期号：　

开始页：　

结束页：　

文献类型：Article; Early Access

关键词：atherosclerosis; endothelial injury; Neu5Ac; sialylation; SQSTM1/p62

摘要："Sialic acid (SIA) has been reported to be a risk factor for atherosclerosis (AS) due to its high plasma levels in such patients. However, the effect of increasing SIA in circulation on endothelial function during AS progression remains unclear. In the present study, ApoE(-/-) mice and endothelial cells line (HUVEC cells) were applied to investigate the effect of SIA on AS progression and its potential molecular mechanism. In vivo, mice were injected intraperitoneally with Neu5Ac (main form of SIA) to keep high-level SIA in circulation. ORO, H&E, and Masson staining were applied to detect the plaque progression. In vitro, HUVECs were treated with Neu5Ac at different times, CCK-8, RT-PCR, western blot, and immunoprecipitation methods were used to analyze its effects on endothelial function and the potential involved mechanism. Results from the present study showed that high plasma levels of Neu5Ac in ApoE(-/-) mice could aggravate the plaque areas as well as increase necrotic core areas and collagen fiber contents. Remarkably, Neu5Ac levels in circulation displayed a positive correlation with AS plaque areas. Furthermore, results from HUVECs showed that Neu5Ac inhibited cells viability in a time/dose-dependent manner, by then induced the activation of inflammation makers such as ICAM-1 and IL-1 beta. Mechanism study showed that the activation of excessive autophagy medicated by SQSTM1/p62 displayed an important role in endothelium inflammatory injury. Neu5Ac could modify SQSTM1/p62 as a sialylation protein, and then increase its level with ubiquitin binding, further inducing ubiquitination degradation and being involved in the excessive autophagy pathway. Inhibition of sialylation by P-3Fax-Neu5Ac, a sialyltransferase inhibitor, reduced the binding of SQSTM1/p62 to ubiquitin. Together, these findings indicated that Neu5Ac increased SQSTM1/p62-ubiquitin binding through sialylation modification, thereby inducing excessive autophagy and subsequent endothelial injury. Inhibition of SQSTM1/p62 sialylation might be a potential strategy for preventing such disease with high levels of Neu5Ac in circulation."

基金机构："We thank all patients and healthy donors who have participated in this study. We are grateful to Associate Professor Jun Zhang (Institute of Life Sciences, Chongqing Medical University) for the gifts of HUVECs mRFP-GFP-LC3 reporter cell line and autophagy"

基金资助正文："We thank all patients and healthy donors who have participated in this study. We are grateful to Associate Professor Jun Zhang (Institute of Life Sciences, Chongqing Medical University) for the gifts of HUVECs mRFP-GFP-LC3 reporter cell line and autophagy antibodies. The authors wish to thank Zhiyi Yuan, Tingting Wang, Wanping Zhang, and Hui Hu for the proofreading and critical revision of the manuscript and Tongchuan Wang, Rong Hu, Zhiwen Dong, and Siyi Yan for the management of laboratory animals."