Highly Up-Regulation of Fas Ligand Gene Expression after Increasing in Oxidized Low-Density Lipoprotein

Abstract

Oxidized low-density lipoprotein (ox-LDL) shows many harmful effects such as induction of apoptosis on function of endothelial cell (EC). Fas ligand (FASL) induces apoptosis in divergent pathological conditions. EC apoptosis which induced by ox-LDL is related with FASL. In this study, we aimed to determine how different ox-LDL levels affect FASL expression in ECs. We treated human umbilical vein endothelial cells (HUVECs) with two different concentration of ox-LDL. FASL gene expression in groups was detected by quantitative polymerase chain reaction (qPCR). We compared FASL gene expression level between the groups according to ∆∆Ct method. FASL gene expression was statistic significantly up-regulated in the group treated higher amounts of ox-LDL (P<0.001). We found that treatment with higher amounts of ox-LDL in HUVECs increased FASL gene expression dramatically. According to our findings, we concluded that increasing the amount of ox-LDL may be critical in inducing the FASL-dependent apoptotic pathway in ECs.

Keywords

• Oxidized low-density lipoprotein
• Fas ligand
• Gene expression
• Quantitative polymerase chain reaction
• Human umbilical vein endothelial cell

References

1. Cao X, Yang J, He L, Liu C. 2024. Circ_0005699 Expedites ox-LDL-Triggered endothelial cell injury via targeting miR-384/ASPH axis. Cardiovasc Toxicol, 24 (10): 1067-1076.
2. Imanishi T, Hano T, Sawamura T, Takarada S, Nishio I. 2002. Oxidized low density lipoprotein potentiation of Fas-induced apoptosis through lectin-like oxidized-low density lipoprotein receptor-1 in human umbilical vascular endothelial cells. Circ J, 66 (11): 1060-1064.
3. Khatana C, Saini NK, Chakrabarti S, Saini V, Sharma A, Saini RV, Saini AK. 2020. Mechanistic insights into the oxidized low-density lipoprotein-induced atherosclerosis. Oxid Med Cell Longev, 5245308.
4. Li D, Yang B, Mehta JL. 1998. Ox-LDL induces apoptosis in human coronary artery endothelial cells: Role of PKC, PTK, bcl-2, and Fas. Am J Physiol, 275: H568-H576.
5. Li HL, Wang AB, Zhang R, Wei YS, Chen HZ, She ZG, Huang Y, Liu DP, Liang CC. 2006. A20 inhibits oxidized low-density lipoprotein-induced apoptosis through negative Fas/Fas ligand-dependent activation of caspase-8 and mitochondrial pathways in murine RAW264.7 macrophages. Cell Physiol, 208: 307-18.
6. Li Q, Wang Y, Li H, Shen G, Hu S. 2014. Ox-LDL influences peripheral Th17/Treg balance by modulating Treg apoptosis and Th17 proliferation in atherosclerotic cerebral infarction. Cell Physiol Biochem, 33(6): 1849-1862.
7. Li X, Yang Y, Wang Z, Jiang S, Meng Y, Song X, Zhao L, Zou L, Li M, Yu T. 2021. Targeting non-coding RNAs in unstable atherosclerotic plaques: Mechanism, regulation, possibilities, and limitations. Int J Biol Sci, 17(13): 3413-3427.
8. Liu Q, Liu Z, Zhou LJ, Cui YL, Xu JM. 2020. The long noncoding RNA NKILA protects against myocardial ischaemic injury by enhancing myocardin expression via suppressing the NF-κB signalling pathway. Exp Cell Res, 15; 387(2): 111774.

9. Livak KJ, Schmittgen TD. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods, 25(4): 402-8.
10. Ma J, Ling J, Tong R, Guo J, Zhu Z. 2024. siLOXL2 inhibits endothelial inflammatory response and EndMT induced by ox-LDL. Cerebrovasc Dis Extra, 14 (1): 165-176.
11. Malarkannan S. 2020. Molecular mechanisms of FASL-mediated ‘reverse-signaling’. Mol Immunol, 127: 31-37.
12. Munno M, Mallia A, Greco A, Modafferi G, Banfi C, Eligini S. 2024. Radical Oxygen species, oxidized low-density lipoproteins, and lectin-like oxidized low-density lipoprotein receptor 1: a vicious circle in atherosclerotic process. Antioxidants, 13 (5): 583.
13. Poznyak AV, Nikiforov NG, Markin AM, Kashirskikh DA, Myasoedova VA, Gerasimova EV, Orekhov AN. 2021. Overview of OxLDL and its impact on cardiovascular health: focus on atherosclerosis. Front Pharmacolü 11: 613780.
14. RT2 Profiler PCR Arrays & Assays Data Analysis Handbook. 2019. https://dataanalysis2.qiagen.com/static/templates/RT2_Profiler_PCR_Data_Analysis_Handbook_190930.pdf (accessed date: November 01, 2024).
15. Sata M, Walsh K. 1998. Oxidized LDL activates Fas-mediated endothelial cell apoptosis. J Clin Invest, 102: 1682-1689.
16. Tian H, Yao ST, Yang NN, Ren J, Jiao P, Zhang X, Qin SC. 2017. D4F alleviates macrophage-derived foam cell apoptosis by inhibiting the NF-κB-dependent Fas/FASL pathway. Sci Rep, 7(1): 7333.
17. Wu MY, Li CJ, Hou MF, and Chu PY. 2017. New insights into the role of inflammation in the pathogenesis of atherosclerosis. Int J Mol Sci, 18 (10): 2034.