Long non-coding RNA alpha-2-macroglobulin antisense RNA 1 regulating oxidized low density lipoprotein-induced human brain microvascular endothelial cell damage by targeting microRNA-106b-5p

LI Wei; WANG Li; WANG Zhi-Hua; LIU Qing-Chun; HAN Rong-Sheng

doi:10.16098/j.issn.0529-1356.2023.03.010

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Acta Anatomica Sinica ›› 2023, Vol. 54 ›› Issue (3) : 319-327. DOI: 10.16098/j.issn.0529-1356.2023.03.010

Long non-coding RNA alpha-2-macroglobulin antisense RNA 1 regulating oxidized low density lipoprotein-induced human brain microvascular endothelial cell damage by targeting microRNA-106b-5p

LI Wei^1* WANG Li² WANG Zhi-hua³ LIU Qing-chun¹ HAN Rong-sheng⁴

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Abstract

Objective To investigate the effect of long non-coding RNA(lncRNA) alpha-2-macroglobulin antisense RNA 1 (A2M-AS1) targeting microRNA (miR）-106b-5p on oxidized low-density lipoprotein (ox-LDL)-induced injury of human brain microvascular endothelial cells. Methods Human brain microvascular endothelial cells (ox-LDL group) were induced by ox-LDL, normal cultured cells were control group (Ctrl); A2M-AS1 overexpression (pcDNA-A2M-AS1 group), empty vector (pcDNA group), miR-106b-5p inhibitor (anti-miR-106b-5p group), negative control (anti-miR-NC group), pcDNA-A2M-AS1 with control mimic NC (miR-NC group), pcDNA-A2M-AS1 with miR-106b-5p mimic (miR-106b-5p mimics group) were transfected into cells and treated with ox-LDL, n=9. Real-time PCR was used to detect the expression levels of A2M-AS1 and miR-106b-5p; Kits were used to detect malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT)); Flow cytometry and TUNEL detected apoptosis; Dual luciferase reporter gene assay detected A2M-AS1 and miR-106b-5p targeting; Western blotting detected Bcl-2 and Bax protein expression. Results Compared with the Ctrl group, the expression level of A2M-AS1 in the ox-LDL group decreased, and the activity of SOD and CAT and the protein level of Bcl-2 decreased (P＜0.05), while the expression level of miR-106b-5p and the level of MDA increased (P<0.05), and the rate of apoptosis and the protein level of Bax increased (P<0.05). Overexpressing A2M-AS1 or interfering with miR-106b-5p decreased the MDA level, apoptosis rate and Bax protein level after ox-LDL-induced cells, and increased SOD, CAT activity and Bcl-2 protein level (P＜0.05). A2M-AS1 targeted miR-106b-5p; up-regulation of miR-106b-5p reversed the effect of overexpressed lncRNA A2M-AS1 on ox-LDL-induced injury of human brain microvascular endothelial cells (P<0.05). Conclusion A2M-AS1 attenuates ox-LDL-induced injury of human brain microvascular endothelial cells by targeting miR-106b-5p.

Key words

Long non-coding RNA

/ Alpha-2-macroglobulin antisense RNA 1 / MicroRNA-106b-5p / Oxidized low density lipoprotein / Oxidative stress / Real-time PCR / Flow cytometry / Human brain microvascular endothelial cell

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LI Wei WANG Li WANG Zhi-hua LIU Qing-chun HAN Rong-sheng.

Long non-coding RNA alpha-2-macroglobulin antisense RNA 1 regulating oxidized low density lipoprotein-induced human brain microvascular endothelial cell damage by targeting microRNA-106b-5p

[J]. Acta Anatomica Sinica. 2023, 54(3): 319-327 https://doi.org/10.16098/j.issn.0529-1356.2023.03.010

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