连接蛋白43在氧化低密度脂蛋白诱导RAW264.7细胞自噬中的作用

doi:10.3969/j.issn.1006-5725.2023.20.003

Abstract

Abstract:

Objective This study aims to explore the role of connexin 43 （Cx43） in the autophagy of RAW264.7 cell induced by oxidized low-density lipoprotein （ox-LDL）. Methods The ox-LDL （100 μg/mL， 24 h） was used to establish the model of autophagy and foam cell formation. RAW264.7 mouse mononuclear macrophage lines were randomly divided into control group， ox-LDL group， ox-LDL+Gap26 group and ox-LDL+Gap19 group. The mRNA levels of Cx43， autophagy protein Beclin-1 and LC3B were detected by qRT-PCR. The protein levels of Cx43， Beclin-1 and LC3B were detected by Western blot. The expression and localization of Cx43， Beclin-1 and LC3B were detected by cellular immunofluorescence. Results The results of oil red O staining showed that red lipid droplets appeared in the cell plasma of the ox-LDL group and cells were foamy 24 h after ox-LDL intervention in RAW264.7 cells. By Western blot， qRT-PCR and immunofluorescence 24 h after ox-LDL intervention in RAW264.7 cells， the expression level of Cx43 in the cells of the ox-LDL group was increased， the expression levels of Beclin-1， LC3B mRNA and protein were decreased in the cells of the ox-LDL group compared with the control group， the expression levels of Beclin-1 and LC3B were decreased in the ox-LDL+Gap26 group compared with the ox-LDL group， and the expression levels of autophagy-related proteins in the ox LDL+Gap19 group were increased compared with the ox-LDL group. Conclusion Cx43 blocking can significantly promote RAW264.7 autophagy induced by ox-LDL.

Key words: Cx43, macrophage, autophagy

CLC Number:

Siyuan XIONG,Lu WANG,Junjie TIAN,Yang GU,Yajing MA,Ketao MA,Yingying ZHANG,Xinzhi. LI. Role of Cx43 in ox⁃LDL induced RAW264.7 autophagy[J]. The Journal of Practical Medicine, 2023, 39(20): 2572-2578.

Figures/Tables 6

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References 25

1	HEDAR AHMED M， STRADNER MARTIN H， ROESSLER A， et al. Autoimmune Rheumatic Diseases and Vascular Function： The Concept of Autoimmune Atherosclerosis［J］. J Clin Med， 2021， 10（19）：4427.
2	CAO H， JIA Q， YAN L， et al. Quercetin Suppresses the Progression of Atherosclerosis by Regulating MST1-Mediated Autophagy in ox-LDL-Induced RAW264.7 Macrophage Foam Cells［J］. Int J Mol Sci， 2019， 20（23）：6093.
3	CHEN W， LI X， GUO S， et al. Tanshinone IIA harmonizes the crosstalk of autophagy and polarization in macrophages via miR-375/KLF4 pathway to attenuate atherosclerosis［J］. Int Immunopharmacol， 2019， 70： 486-497.
4	CONWAY O， AKPINAR H A， ROGOV V V， et al. Selective Autophagy Receptors in Neuronal Health and Disease［J］. J Mol Biol， 2020， 432（8）： 2483-2509.
5	PURI D， SUBRAMANYAM D. Stress - （self） eating： Epigenetic regulation of autophagy in response to psychological stress［J］.FEBS J， 2019， 286（13）： 2447-2460.
6	GAO J， CHENG T， QIN A， et al. Glucocorticoid impairs cell-cell communication by autophagy-mediated degradation of connexin 43 in osteocytes［J］. Oncotarget， 2016， 7（19）： 26966-26978.
7	CAO Q， DU H， FU X， et al. Artemisinin Attenuated Atherosclerosis in High-Fat Diet-Fed ApoE-/- Mice by Promoting Macrophage Autophagy Through the AMPK/mTOR/ULK1 Pathway［J］. Cardiovasc Pharmacol， 2020， 75（4）： 321-332.
8	QUAN M， LV H， LIU Z， et al. MST1 Suppresses Disturbed Flow Induced Atherosclerosis［J］. Circ Res， 2022， 131（9）： 748-764.
9	COMARIŢA I K， VÎLCU A， CONSTANTIN A， et al. Therapeutic Potential of Stem Cell-Derived Extracellular Vesicles on Atherosclerosis-Induced Vascular Dysfunction and Its Key Molecular Players［J］. Front Cell Dev Biol， 2022， 10： 817180.
10	SÁEZ J C， CONTRERAS-DUARTE S， LABRA V C， et al. Interferon-γ and high glucose-induced opening of Cx43 hemichannels causes endothelial cell dysfunction and damage［J］. Biochim Biophys Acta Mol Cell Res， 2020， 1867（8）： 118720.
11	RAMADAN R， VROMANS E， ANANG D C， et al. Connexin43 Hemichannel Targeting With TAT-Gap19 Alleviates Radiation-Induced Endothelial Cell Damage［J］. Front Pharmacol， 2020， 11： 212.
12	LI M， WANG Z， SHAO J， et al. Captopril Attenuates the Upregulated Connexin 43 Expression in Artery Calcification［J］.Arch Med Res， 2020， 51（3）： 215-223.
13	KOELWYN G J， CORR E M， ERBAY E， et al. Regulation of macrophage immunometabolism in atherosclerosis［J］. Nat Immunol， 2018， 19（6）： 526-537.
14	PENG X， CHEN H， LI Y， et al. Effects of NIX-mediated mitophagy on ox-LDL-induced macrophage pyroptosis in atherosclerosis［J］. Cell Biol Int， 2020， 44（7）： 1481-1490.
15	YANG L， LI T， ZHA L. Foxc2 Alleviates Ox-LDL-Induced Lipid Accumulation， Inflammation， and Apoptosis of Macrophage via Regulating the Expression of Angptl2［J］. Inflammation， 2020， 43（4）： 1397-1410.
16	XUE Z， YUAN W， LI J， et al. Cyclophilin A mediates the ox-LDL-induced activation and apoptosis of macrophages via autophagy［J］. Int J Cardiol， 2017， 230： 142-148.
17	左夏林，唐艳艳，李孔平，等.Cx43缝隙连接蛋白羧基末端磷酸化修饰对其功能的调控及其在脑缺血中的研究进展［J］. 实用医学杂志，2021，37（21）：2804-2808.
18	刘卫东，孔文洁，高峰.缝隙连接蛋白调节机制的研究进展［J］.生理科学进展，2021，52（4）：275-279.
19	王文博，荣毅，陈玲，等. Cx43调控NLRP3炎性小体参与α1-AR激活诱导的心脏急性交感应激［J］. 安徽医科大学学报，2022，57（4）：534-539.
20	周高生，姚华国，邓烈华. 脓毒症时血管内皮细胞连接变化的研究进展［J］. 广东医学院学报，2013，31（3）：319-321.
21	吴玲玲，何芳. 缝隙连接在血管内皮和平滑肌细胞中的作用及其与高血压的关系［J］. 国际病理科学与临床杂志，2013，33（4）：332-337.
22	SHEN C， CHEN J， LEE Y， et al. mTOR- and SGK-Mediated Connexin 43 Expression Participates in Lipopolysaccharide-Stimulated Macrophage Migration through the iNOS/Src/FAK Axis［J］. J Immunol， 2018， 201（10）： 2986-2997.
23	GAO J， CHENG T S， QIN A， et al. Correction： Glucocorticoid impairs cell-cell communication by autophagy-mediated degradation of connexin 43 in osteocytes［J］. Oncotarget， 2019， 10（40）： 4079.
24	林张军，李倩，芮耀诚. Nogo-B抑制Ox-LDL诱导的巨噬细胞泡沫化［J］. 中国生物化学与分子生物学报， 2019， 35（11）： 1225-1233.
25	陈方，吴小枫，林珍，等. 油酰乙醇胺抑制巨噬细胞泡沫化改善动脉粥样硬化的机制［J］. 免疫学杂志，2022，38（12）：1021-1029.

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Role of Cx43 in ox⁃LDL induced RAW264.7 autophagy

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