[Gly14]-humanin通过FPRL1修复Cathepsin D功能,促进HUVECs中ox-LDL的自噬性降解
首发时间:2019-05-15
摘要:血管内皮细胞中的氧化低密度脂蛋白(ox-LDL)的异常聚集是动脉粥样硬化病变的主要病理变化之一。我们之前研究表明,外源性给予S14G-HN(HNG)可以通过抑制入胞,促进外排,减少ox-LDL在血管内皮细胞的聚集。在该研究中,流式细胞仪检测细胞内dil标记的ox-LDL(Dil-ox-LDL)含量及脂质定量试剂盒检测结果显示,ox-LDL(0, 30 μg/mL,60 μg/mL,90 μg/mL,120 μg/mL)能呈浓度依赖性引起HUVECs内脂质和胆固醇的聚集。Western blotting结果表明,ox-LDL以浓度依赖性方式增加自噬标志性蛋白P62和LC3-II的表达。Cathepsin D活性试剂盒结果显示ox-LDL暴露会抑制细胞Cathepsin D的功能。HNG预处理能够降低ox-LDL引起的HUVECs中脂质和胆固醇聚集,增加细胞LC3-II蛋白水平,减少p62蛋白含量,并修复ox-LDL诱导的Cathepsin D功能受损。用FPRL1 siRNA和FPRL1特异性抑制剂Boc-MLF抑制FPRL1途径,会抑制HNG的上述所有保护作用。这些结果表明,Humanin抑制ox-LDL诱导的HUVECs内脂质聚集作用还与其改善受损的Cathepsin D酶活性,进而修复HUVECs中由ox-LDL引起的自噬流损伤,促进脂质的自噬性降解有关。
关键词: 神经生物学 HNG Ox-LDL FPRL1 HUVECs Cathepsin D
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[Gly14]-humanin repairs cathepsin D function by FPRL1 and promotes autophagic degradation of ox-LDL in HUVECs
Abstract:Abnormal aggregation of oxidized low density lipoprotein (ox-LDL) in vascular endothelial cells is one of the major pathological changes in atherosclerotic lesions. Our previous studies have shown that exogenous administration of S14G-HN (HNG) can promote efflux and reduce ox-LDL accumulation in vascular endothelial cells by inhibiting cell entry. In this study, flow cytometry was used to detect intracellular dil-labeled ox-LDL (Dil-ox-LDL) levels and lipid quantification kits showed ox-LDL (0, 30 μg/mL, 60 μg/ mL, 90 μg/mL, 120 μg/mL) caused concentration of lipids and cholesterol in HUVECs in a concentration-dependent manner. Western blotting results showed that ox-LDL increased the expression of autophagy marker proteins P62 and LC3-II in a concentration-dependent manner. The Cathepsin D activity kit showed that ox-LDL inhibited the function of Cathepsin D. HNG pretreatment could reduce lipid and cholesterol aggregation in HUVECs induced by ox-LDL, increase LC3-II protein level, decrease p62 protein content, and repair ox-LDL-induced Cathepsin D function impaired. Inhibition of the FPRL1 pathway by FPRL1 siRNA and the FPRL1 specific inhibitor Boc-MLF, all protections of HNG were blocked. These results indicated that Humanin could repair Cathepsin D activity and protein levels in HUVECs to repair lysosomal functional damage induced by ox-LDL, further repair ox-LDL-induced autophagic damage in HUVECs.
Keywords: Neurobiology HNG Ox-LDL FPRL1 HUVECs Cathepsin D
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[Gly14]-humanin通过FPRL1修复Cathepsin D功能,促进HUVECs中ox-LDL的自噬性降解
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