Oxidized low-density lipoprotein (ox-LDL) could induce endothelial injury and played a vital role in the progression and development of atherosclerosis. This study aimed to investigate the role of Opa-interacting protein 5 antisense RNA 1 (OIP5-AS1) in ox-LDL-induced human umbilical vascular endothelial cells (HUVECs) injury and the potential mechanisms.
METHODS:
Cell proliferation and apoptosis were evaluated by Cell Counting Kit-8 (CCK-8) assay and flow cytometry assay, respectively. The levels of lactate dehydrogenase (LDH), reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD) and nitric oxide (NO) were detected by corresponding detection kits, respectively. Quantitative real-time PCR (qRT-PCR) was conducted to measure the expression of OIP5-AS1 or microRNA-30c-5p (miR-30c-5p) in HUVECs. Binding between OIP5-AS1 and miR-30c-5p was predicted through bioinformatics analysis and confirmed by dual-luciferase reporter assay and RNA immunoprecipitation (RIP). Western blot was used to analyze p-IκB, IκB, p-p65 and p65 levels.
RESULTS:
In HUVECs, exposure to ox-LDL led to a decrease in cell viability and an increase in LDH release and apoptosis with concomitant enhancement of oxidative stress, as evidenced by increased ROS and MDA generation, as well as decreased SOD activity and NO levels, while OIP5-AS1 knockdown or miR-30c-5p upregulation could rescue these effects above. Mechanically, OIP5-AS1 functioned as a sponge of miR-30c-5p. OIP5-AS1-induced injury and apoptosis, oxidative stress and activation of NF-κB pathway were reversed by miR-30c-5p in ox-LDL-treated HUVECs.
CONCLUSION:
OIP5-AS1 contributed to ox-LDL-treated HUVECs injury by activation of NF-κB pathway via miR-30c-5p.
Global Burden of Disease Study C. Global, regional, and national incidence, prevalence, and years lived with disability for 301 acute and chronic diseases and injuries in 188 countries, 1990-2013: a systematic analysis for the Global Burden of Disease Study 2013. Lancet. 2015;386:743–800.
2.
BadimonL, VilahurG. Thrombosis formation on atherosclerotic lesions and plaque rupture. J Intern Med. 2014;276:618–32.
DruryMI, SweeneyEC, BarragryJ, MackeyJ. Localised (pretibial) myxoedema in hyperthyroidism: a report of six cases. J Ir Med Assoc. 1971;64:623–6.
5.
Krüger-GengeA, BlockiA, FrankeRP, JungF. Vascular Endothelial Cell Biology: An Update. Int J Mol Sci. 2019;20.
6.
LibbyP, RidkerPM, MaseriA. Inflammation and atherosclerosis. Circulation. 2002;105:1135–43.
7.
ShenZ, LiH. Long non-coding RNA GAS5 knockdown facilitates proliferation and impedes apoptosis by regulating miR-128-3p/FBLN2 axis in ox-LDL-induced THP-1 cells. Clin Hemorheol Microcirc. 2020.
8.
PirilloA, NorataGD, CatapanoAL. LOX-1, OxLDL, and atherosclerosis. Mediators Inflamm. 2013;2013:152786.
9.
ThakkarS, WangX, KhaidakovM, DaiY, GokulanK, MehtaJL, et al. Structure-based Design Targeted at LOX-1, a Receptor for Oxidized Low-Density Lipoprotein. Sci Rep. 2015;5:16740.
10.
WuJ, ZhaoW, WangZ, XiangX, ZhangS, LiuL. Long non-coding RNA SNHG20 promotes the tumorigenesis of oral squamous cell carcinoma via targeting miR-197/LIN28 axis. J Cell Mol Med. 2019;23:680–8.
11.
ZhangX, YangL, XuG. Silencing of long noncoding RNA TUG1 inhibits viability and promotes apoptosis of acute myeloid leukemia cells by targeting microRNA-221-3p/KIT axis. Clin Hemorheol Microcirc. 2020;76:425–37.
12.
AryalB, SuarezY. Non-coding RNA regulation of endothelial and macrophage functions during atherosclerosis. Vascul Pharmacol. 2019;114:64–75.
13.
SallamT, SandhuJ, TontonozP. Long Noncoding RNA Discovery in Cardiovascular Disease: Decoding Form to Function. Circ Res. 2018;122:155–66.
14.
ZhouT, DingJW, WangXA, ZhengXX. Long noncoding RNAs and atherosclerosis. Atherosclerosis. 2016;248:51–61.
15.
LiM, DuanL, LiY, LiuB. Long noncoding RNA/circular noncoding RNA-miRNA-mRNA axes in cardiovascular diseases. Life Sci. 2019;233:116440.
16.
ZhangBY, JinZ, ZhaoZ. Long intergenic noncoding RNA 00305 sponges miR-136 to regulate the hypoxia induced apoptosis of vascular endothelial cells. Biomed Pharmacother. 2017;94:238–43.
17.
HalimulatiM, DumanB, NijiatiJ, AizeziA. Long noncoding RNA TCONS_00024652 regulates vascular endothelial cell proliferation and angiogenesis via microRNA-21. Exp Ther Med. 2018;16:3309–16.
18.
WangM, LiuY, LiC, ZhangY, ZhouX, LuC. Long noncoding RNA OIP5-AS1 accelerates the ox-LDL mediated vascular endothelial cells apoptosis through targeting GSK-3beta via recruiting EZH2. Am J Transl Res. 2019;11:1827–34.
19.
ZhaoZ, VajenT, KarshovskaE, DickhoutA, SchmittMM, MegensRTA, et al. Deletion of junctional adhesion molecule A from platelets increases early-stage neointima formation after wire injury in hyperlipidemic mice. J Cell Mol Med. 2017;21:1523–31.
20.
VenugopalP, KoshyT, LavuV, Ranga RaoS, RamasamyS, HariharanS, et al. Differential expression of microRNAs let-7a, miR-125b, miR-100, and miR-21 and interaction with NF-kB pathway genes in periodontitis pathogenesis. J Cell Physiol. 2018;233:5877–84.
21.
LiuAH, WuYT, WangYP. MicroRNA-129-5p inhibits the development of autoimmune encephalomyelitis-related epilepsy by targeting HMGB1 through the TLR4/NF-kB signaling pathway. Brain Res Bull. 2017;132:139–49.
22.
KagiyaT. MicroRNAs: Potential Biomarkers and Therapeutic Targets for Alveolar Bone Loss in Periodontal Disease. Int J Mol Sci. 2016;17.
LiuC, PanC, CaiY, WangH. Interplay Between Long Noncoding RNA ZEB1-AS1 and miR-200s Regulates Osteosarcoma Cell Proliferation and Migration. Journal of cellular biochemistry. 2017;118:2250–60.
25.
ShiJ, WangH, FengW, HuangS, AnJ, QiuY, et al. Long non-coding RNA HOTTIP promotes hypoxia-induced glycolysis through targeting miR-615-3p/HMGB3 axis in non-small cell lung cancer cells. European journal of pharmacology. 2019;862:172615.
26.
ZhangM, XueY, ChenH, MengL, ChenB, GongH, et al. Resveratrol Inhibits MMP3 and MMP9 Expression and Secretion by Suppressing TLR4/NF-κB/STAT3 Activation in Ox-LDL-Treated HUVECs. Oxidative medicine and cellular longevity. 2019;2019.
27.
GuoF, TangC, LiY, LiuY, LvP, WangW, et al. The interplay of LncRNA ANRIL and miR-181b on the inflammation-relevant coronary artery disease through mediating NF-kappaB signalling pathway. J Cell Mol Med. 2018;22:5062–75.
28.
LiS, SunY, ZhongL, XiaoZ, YangM, ChenM, et al. The suppression of ox-LDL-induced inflammatory cytokine release and apoptosis of HCAECs by long non-coding RNA-MALAT1 via regulating microRNA-155/SOCS1 pathway. Nutr Metab Cardiovasc Dis. 2018;28:1175–87.
29.
LiuJ, HuangGQ, KeZP. Silence of long intergenic noncoding RNA HOTAIR ameliorates oxidative stress and inflammation response in ox-LDL-treated human macrophages by upregulating miR-330-5p. J Cell Physiol. 2019;234:5134–42.
30.
LiY, HanX, FengH, HanJ. Long noncoding RNA OIP5-AS1 in cancer. Clin Chim Acta. 2019;499:75–80.
31.
WangY, ShiF, XiaY, ZhaoH. LncRNA OIP5-AS1 predicts poor prognosis and regulates cell proliferation and apoptosis in bladder cancer. J Cell Biochem. 2018.
32.
ZouY, YaoS, ChenX, LiuD, WangJ, YuanX, et al. LncRNA OIP5-AS1 regulates radioresistance by targeting DYRK1A through miR-369-3p in colorectal cancer cells. Eur J Cell Biol. 2018;97:369–78.
33.
BaiY, LiS. Long noncoding RNA OIP5-AS1 aggravates cell proliferation, migration in gastric cancer by epigenetically silencing NLRP6 expression via binding EZH2. J Cell Biochem. 2020;121:353–62.
34.
WangM, SunX, YangY, JiaoW. Long non-coding RNA OIP5-AS1 promotes proliferation of lung cancer cells and leads to poor prognosis by targeting miR-378a-3p. Thorac Cancer. 2018;9:939–49.
35.
YangS, SunJ. LncRNA SRA deregulation contributes to the development of atherosclerosis by causing dysfunction of endothelial cells through repressing the expression of adipose triglyceride lipase. Mol Med Rep. 2018;18:5207–5214.
36.
YariM, BitarafanS, BroumandMA, FazeliZ, RahimiM, GhaderianSMH, et al. Association between Long Noncoding RNA ANRIL Expression Variants and Susceptibility to Coronary Artery Disease. Int J Mol Cell Med. 2018;7:1–7.
37.
ZhangM, XueY, ChenH, MengL, ChenB, GongH, et al. Resveratrol Inhibits MMP3 and MMP9 Expression and Secretion by Suppressing TLR4/NF-κB/STAT3 Activation in Ox-LDL-Treated HUVECs. Oxid Med Cell Longev. 2019;2019:9013169.
38.
ZhongX, ZhangL, LiY, LiP, LiJ, ChengG. Kaempferol alleviates ox-LDL-induced apoptosis by up-regulation of miR-26a-5p via inhibiting TLR4/NF-κB pathway in human endothelial cells. Biomedicine & Pharmacotherapy. 2018;108:1783–9.
39.
YuanX, ChenJ, DaiM. Paeonol promotes microRNA-126 expression to inhibit monocyte adhesion to ox-LDL-injured vascular endothelial cells and block the activation of the PI3K/Akt/NF-κB pathway. International Journal of Molecular Medicine. 2016;38:1871–8.
40.
YangL, GaoC. MiR-590 Inhibits Endothelial Cell Apoptosis by Inactivating the TLR4/NF-κB Pathway in Atherosclerosis. Yonsei Medical Journal. 2019;60:298–307.
