[Gly14]-humanin restores cathepsin D function via FPRL1 and promotes autophagic degradation of Ox-LDL in HUVECs
[Gly14]-humanin restores cathepsin D function via FPRL1 and promotes autophagic degradation of Ox-LDL in HUVECs
复制标题
[Gly14]-人素通过 FPRL1 恢复组织蛋白酶 D 功能并促进 HUVEC 中 Ox-LDL 的自噬降解
DOI：
10.1016/j.numecd.2020.07.022
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发表时间：
2020-11-27
影响因子：
3.9
通讯作者：
Zhang, Yanlin
中科院分区：
文献类型：
--
作者：
Ding, Yu;Feng, Yue;Zhang, Yanlin
背景与目的：氧化低密度脂蛋白在血管内皮细胞中的异常聚集是动脉粥样硬化病变的主要病理改变之一。本研究旨在探讨人蛋白(HN)促进人脐静脉内皮细胞(HUVEC)氧化低密度脂蛋白(Ox-LDL)自噬降解的机制。方法与结果：流式细胞术和脂质定量结果显示，Ox-LDL可引起HUVEC中脂质和胆固醇的积聚。Western印迹结果显示，Ox-LDL以浓度依赖的方式增加自噬相关蛋白P62和Lc3-II的表达。组织蛋白酶D活性测定显示，Ox-LDL可抑制组织蛋白酶D的功能，HNG可减少Ox-LDL诱导的HUVEC的脂质和胆固醇聚集，增加Lc3-II蛋白水平，降低p62蛋白含量，并逆转Ox-LDL诱导的组织蛋白酶D功能损害。用FPRL1 siRNA或FPRL1特异性抑制剂Boc-MLF抑制FPRL1通路可阻断HNG的所有保护作用。提示HNG可通过其膜蛋白受体FPRL1恢复Ox-LDL诱导的HUVECs组织蛋白酶D活性和蛋白水平，修复Ox-LDL诱导的HUVEC溶酶体功能损伤，进而修复Ox-LDL诱导的HUVECs自噬损伤。结论：HNG通过其膜蛋白受体FPRL1恢复Ox-LDL诱导的HUVECs组织蛋白酶D活性，促进Ox-LDL的自噬降解。(C)2020年意大利糖尿病学会、意大利动脉粥样硬化研究学会、意大利人类营养学会和费德里科二世大学临床内外科。爱思唯尔出版，版权所有。
Background and aim: Abnormal aggregation of oxidized low-density lipoprotein (OxLDL) in vascular endothelial cells (VECs) is one of the major pathological changes in atherosclerotic lesions. Our research aimed to assess the mechanism of humanin (HN) in promoting autophagic degradation of Ox-LDL in HUVECs.Methods and results: Flow cytometry and lipid quantitation results showed that Ox-LDL caused lipid and cholesterol accumulation in HUVECs. Western blot results showed that Ox-LDL increased the expression of autophagy-related proteins P62 and LC3-II in a concentrationdependent manner. The cathepsin D activity assay showed that Ox-LDL inhibited the function of cathepsin D. HNG pretreatment reduced lipid and cholesterol aggregation in HUVECs induced by Ox-LDL, increased LC3-II protein level, decreased P62 protein content, and reversed Ox-LDLinduced cathepsin D functional impairment. Inhibition of the FPRL1 pathway by FPRL1 siRNA or the FPRL1-specific inhibitor Boc-MLF blocked all HNG's protective effects. These results indicate that HNG could restore cathepsin D activity and protein level in HUVECs to repair lysosomal functional damage induced by Ox-LDL, further repairing Ox-LDL-induced autophagic damage in HUVECs.Conclusion: HNG restores the activity of Ox-LDL-induced damaged lysosomal enzyme cathepsin D through its membrane protein receptor FPRL1 to promote autophagic degradation of Ox-LDL in HUVECs. (C) 2020 The Italian Diabetes Society, the Italian Society for the Study of Atherosclerosis, the Italian Society of Human Nutrition and the Department of Clinical Medicine and Surgery, Federico II University. Published by Elsevier B.V. All rights reserved.