PCSK9/LOX-1 is associated with T2DM and regulates high glucose-induced lipid metabolism dysfunction in human microvascular endothelial cells
Abstract
Introduction: The proprotein convertase subtilisin/kexin type 9/lectin-like oxidized low-density lipoprotein receptor-1 (PCSK9/LOX-1) axis plays a crucial role in regulating vascular endothelial cell function, but its specific involvement in type 2 diabetes mellitus (T2DM) remains unclear. This study aims to explore the potential mechanism of the PCSK9/LOX-1 axis in high-glucose (HG)-induced vascular endothelial cell dysfunction.
Material and methods: Peripheral blood samples were collected from T2DM patients to analyse the correlation between PCSK9 and blood lipid levels. Human microvascular endothelial cells (HMEC-1) exposed to high glucose concentration were used as a model of diabetic= angiopathy (DA). Levels of PCSK9, reactive oxygen species (ROS), malonodialdehyde (MDA), interleukins (IL): IL-6, IL-1β, superoxide dismutase (SOD), and tumour necrosis factor alpha (TNF-α) were determined by enzyme-linked immunosorbent assay (ELISA) and biochemical methods. Additionally, intracellular total cholesterol (TC) and cholesterol ester (CE) levels were detected using enzyme chemistry. Expression of PCSK9 and LOX-1 was assessed through real-time quantitative polymerase chain reaction (RT-qPCR) and western blotting.
Results: Compared to the normal group, PCSK9 levels were significantly elevated in T2DM patients. Furthermore, PCSK9 levels were found to be positively correlated with body mass index (BMI), waistline, triglyceride (TG), cholesterol, low-density lipoprotein cholesterol (LDL-C), glycated hemoglobin (HbAlc), and intracardiac fat pad levels in T2DM patients. HG exposure led to reduced activity of HMEC-1 cells, along with increased levels of apoptosis, oxidative stress, and inflammation, all of which were counteracted by si-PCSK9. The inhibitory effects of si-PCSK9 on LOX-1 expression, as well as TC and CE contents in HMEC-1 cells induced by HG, were observed. Moreover, intervention with oe-LOX-1 reversed the effects of si-PCSK9 in HG-induced HMEC-1 cells.
Conclusion: Silencing of PCSK9 inhibited HG-induced inflammation, oxidative stress, and lipid metabolic dysfunction in HMEC-1 cells via LOX-1.
Keywords: T2DMdiabetic angiopathyPCSK9lipid metabolismHMEC-1 cells
References
- Athyros VG, Doumas M, Imprialos KP, et al. Diabetes and lipid metabolism. Hormones (Athens). 2018; 17(1): 61–67.
- Kahn SE, Cooper ME, Del Prato S. Pathophysiology and treatment of type 2 diabetes: perspectives on the past, present, and future. Lancet. 2014; 383(9922): 1068–1083.
- Prattichizzo F, Giuliani A, Sabbatinelli J, et al. Prevalence of residual inflammatory risk and associated clinical variables in patients with type 2 diabetes. Diabetes Obes Metab. 2020; 22(9): 1696–1700.
- Defronzo RA. Banting Lecture. From the triumvirate to the ominous octet: a new paradigm for the treatment of type 2 diabetes mellitus. Diabetes. 2009; 58(4): 773–795.
- Prattichizzo F, La Sala L, Ceriello A. Two drugs are better than one to start T2DM therapy. Nat Rev Endocrinol. 2020; 16(1): 15–16.
- Yin Z, Wang X, Yang X, et al. Salvia miltiorrhiza in Anti-diabetic Angiopathy. Curr Mol Pharmacol. 2021; 14(6): 960–974.
- Xiang H, Song R, Ouyang J, et al. Organelle dynamics of endothelial mitochondria in diabetic angiopathy. Eur J Pharmacol. 2021; 895: 173865.
- Li L, Sawamura T, Renier G. Glucose enhances endothelial LOX-1 expression: role for LOX-1 in glucose-induced human monocyte adhesion to endothelium. Diabetes. 2003; 52(7): 1843–1850.
- Kattoor AJ, Goel A, Mehta JL. LOX-1: Regulation, Signaling and Its Role in Atherosclerosis. Antioxidants (Basel). 2019; 8(7).
- Ding Z, Liu S, Wang X, et al. Oxidant stress in mitochondrial DNA damage, autophagy and inflammation in atherosclerosis. Sci Rep. 2013; 3: 1077.
- Ding Z, Liu S, Wang X, et al. Hemodynamic shear stress via ROS modulates PCSK9 expression in human vascular endothelial and smooth muscle cells and along the mouse aorta. Antioxid Redox Signal. 2015; 22(9): 760–771.
- Peng J, Zhu CG, Li JJ. The predictive utility of circulating PCSK9 levels on diabetes mellitus. Cardiovasc Diabetol. 2021; 20(1): 45.
- Seidah NG, Awan Z, Chrétien M, et al. PCSK9: a key modulator of cardiovascular health. Circ Res. 2014; 114(6): 1022–1036.
- Li Q, Xuan W, Jia Z, et al. HRD1 prevents atherosclerosis-mediated endothelial cell apoptosis by promoting LOX-1 degradation. Cell Cycle. 2020; 19(12): 1466–1477.
- Raheem Lateef Al-Awsi G, Hadi Lafta M, Hashim Kzar H, et al. PCSK9 pathway-noncoding RNAs crosstalk: Emerging opportunities for novel therapeutic approaches in inflammatory atherosclerosis. Int Immunopharmacol. 2022; 113(Pt A): 109318.
- Ren Yi, Xie W, Yang S, et al. Angiotensin-converting enzyme 2 inhibits inflammation and apoptosis in high glucose-stimulated microvascular endothelial cell damage by regulating the JAK2/STAT3 signaling pathway. Bioengineered. 2022; 13(4): 10802–10810.
- Han YM, Yang H, Huang QL, et al. Risk prediction of diabetes and pre-diabetes based on physical examination data. Math Biosci Eng. 2022; 19(4): 3597–3608.
- Gamble W, Vaughan M, Kruth HS, et al. Procedure for determination of free and total cholesterol in micro- or nanogram amounts suitable for studies with cultured cells. J Lipid Res. 1978; 19(8): 1068–1070.
- Sun S, Cheng B, Wu X, et al. Chlamydia pneumoniae disrupts lipid metabolism in human umbilical vein endothelial cells. Mol Med Rep. 2014; 10(2): 1150–1156.
- Da Dalt L, Ruscica M, Bonacina F, et al. PCSK9 deficiency reduces insulin secretion and promotes glucose intolerance: the role of the low-density lipoprotein receptor. Eur Heart J. 2019; 40(4): 357–368.
- Sabatine MS. PCSK9 inhibitors: clinical evidence and implementation. Nat Rev Cardiol. 2019; 16(3): 155–165.
- Guo Y, Yan B, Tai S, et al. PCSK9: Associated with cardiac diseases and their risk factors? Arch Biochem Biophys. 2021; 704: 108717.
- Cammisotto V, Baratta F, Simeone PG, et al. Proprotein Convertase Subtilisin Kexin Type 9 (PCSK9) Beyond Lipids: The Role in Oxidative Stress and Thrombosis. Antioxidants (Basel). 2022; 11(3).
- Yan Li, Jia Q, Cao H, et al. Fisetin ameliorates atherosclerosis by regulating PCSK9 and LOX-1 in apoE mice. Exp Ther Med. 2021; 21(1): 25.
- Katsuki S, K Jha P, Lupieri A, et al. Proprotein Convertase Subtilisin/Kexin 9 (PCSK9) Promotes Macrophage Activation via LDL Receptor-Independent Mechanisms. Circ Res. 2022; 131(11): 873–889.
- Wu NQ, Shi HW, Li JJ. Proprotein Convertase Subtilisin/Kexin Type 9 and Inflammation: An Updated Review. Front Cardiovasc Med. 2022; 9: 763516.
- Talasaz AH, Ho ACJ, Bhatty F, et al. Meta-analysis of clinical outcomes of PCSK9 modulators in patients with established ASCVD. Pharmacotherapy. 2021; 41(12): 1009–1023.
- Wiciński M, Żak J, Malinowski B, et al. PCSK9 signaling pathways and their potential importance in clinical practice. EPMA J. 2017; 8(4): 391–402.
- Momtazi-Borojeni AA, Jaafari MR, Abdollahi E, et al. Impact of PCSK9 Immunization on Glycemic Indices in Diabetic Rats. J Diabetes Res. 2021; 2021: 4757170.
- Ding Z, Liu S, Wang X, et al. Cross-talk between LOX-1 and PCSK9 in vascular tissues. Cardiovasc Res. 2015; 107(4): 556–567.
- Ding Z, Pothineni NV, Goel A, et al. PCSK9 and inflammation: role of shear stress, pro-inflammatory cytokines, and LOX-1. Cardiovasc Res. 2020; 116(5): 908–915.
- Liu W, He P, Cheng B, et al. Chlamydia pneumoniae disturbs cholesterol homeostasis in human THP-1 macrophages via JNK-PPARγ dependent signal transduction pathways. Microbes Infect. 2010; 12(14-15): 1226–1235.
- Cupido AJ, Reeskamp LF, Hingorani AD, et al. Joint Genetic Inhibition of PCSK9 and CETP and the Association With Coronary Artery Disease: A Factorial Mendelian Randomization Study. JAMA Cardiol. 2022; 7(9): 955–964.
- Tam J, Thankam F, Agrawal DK, et al. Critical Role of LOX-1-PCSK9 Axis in the Pathogenesis of Atheroma Formation and Its Instability. Heart Lung Circ. 2021; 30(10): 1456–1466.
- Huang YP, Wang YS, Liu YY, et al. Chemical Characterization and Atherosclerosis Alleviation Effects of Gypenosides from through Ameliorating Endothelial Dysfunction via the PCSK9/LOX-1 Pathway. J Agric Food Chem. 2022; 70(38): 11944–11957.
- Li W, Park H, Guo E, et al. Aerobic Exercise Training Inhibits Neointimal Formation via Reduction of PCSK9 and LOX-1 in Atherosclerosis. Biomedicines. 2020; 8(4).
- Zhang Y, Gu Y, Chen Y, et al. Dingxin Recipe IV attenuates atherosclerosis by regulating lipid metabolism through LXR-α/SREBP1 pathway and modulating the gut microbiota in ApoE mice fed with HFD. J Ethnopharmacol. 2021; 266: 113436.
