经靶向OPN激活的IKBα/NF-κB信号通路对细胞自噬、细胞增殖和细胞周期的影响  

Effects of IKBα/NF-κB signaling pathway(activated by trans-targeted osteopontin)on cell autophagy,proliferation and cell cycle

作　　者：周锐 刘川川 刘瑞欣[1] 刘辉琦[1] 韩莹 马兰[3] 王生兰[1] ZHOU Rui;LIU Chuanchuan;LIU Ruixin;LIU Huiqi;HAN Ying;MA Lan;WANG Shenglan(Medical Department of Qinghai University,Xining 810001,China;Key Laboratory of Hydatid Disease,Qinghai University Affiliated Hospital,Xining 810001,China;Research Center for High Altitude Medicine,Qinghai University,Xining 810001,China)

机构地区：[1]青海大学医学部,西宁810001 [2]青海大学附属医院包虫病重点实验室,西宁810001 [3]青海大学高原医学研究中心,西宁810001

出　　处：《中国高原医学与生物学杂志》2024年第3期145-155,共11页Journal of Chinese High Altitude Medicine & Biology

基　　金：国家自然科学基金项目(8186006);青海省科技厅自然科学基金项目(2021-ZJ-738);青海大学青年科研基金项目(2023-QYY-4)。

摘　　要：目的探讨经靶向骨桥蛋白(OPN)激活的IKBα/NF-κB信号通路对低氧环境中肺动脉平滑肌细胞(PASMCs)的自噬、增殖和周期的影响。方法培养大鼠PASMCs;观察低氧组PASMCs中OPN的表达情况和IKBα/NF-κB的表达情况。将带有OPN(过表达或敲低)的慢病毒与PASMCs共培养,在OPN过表达的PASMCs中加入NF-κB抑制剂QNZ干扰IKBα/NF-κB信号通路,随后在基因和蛋白层面观察OPN对自噬的影响,并在电镜下观察自噬体数量。用5-乙炔基-2-脱氧嘧啶核苷法和流式细胞术检测OPN对细胞增殖能力和细胞周期的影响。结果低氧组PASMCs表达的OPN高于常氧组(P<0.05),低氧组NF-κB高于常氧组(P<0.05)。常氧组可通过PASMCs过表达OPN激活NF-κB来抑制自噬蛋白LC3B、Beclin1,加入NF-κB抑制剂后可以促进PASMCs的自噬蛋白表达(P<0.05);干扰低氧组PASMCs中OPN的表达可以进一步促进自噬蛋白和自噬小体的表达(P<0.05)。抑制PASMCs中OPN和NF-κB的表达可以阻断细胞从G0/G1期向S期过渡并减弱由OPN增多引起的增殖(P<0.05)。结论经靶向OPN可以激活IKBα/NF-κB信号通路;激活的IKBα/NF-κB信号通路对低氧环境中PASMCs的自噬、增殖和细胞周期产生影响。Objective To explore the effects of IKBα/NF-κB signaling pathway activated by trans-targeted osteopontin(OPN)on the cell autophagy,proliferation and cell cycle of pulmonary artery smooth muscle cells(PASMCs)in hypoxic environment.Methods Rats PASMCs were cultured.The expressions of OPN and IKBα/NF-κB were observed in the PASMCs of hypoxia group.Lentiviruses with OPN(overexpressed or knocked down)were co-cultured with PASMCs,and the NF-κB inhibitor QNZ was added to PASMCs with OPN overexpression to interfere with the IKBα/NF-κB signaling pathway.The effects of OPN on autophagy were observed at the gene and protein levels.The number of autophagosomes was observed under electron microscopy.The effects of OPN on the cell proliferation ability and cell cycle were detected by 5-ethynyl-2-deoxypyrimidine nucleoside assay and flow cytometry.Results PASMCs in the hypoxia group expressed higher OPN than the normoxia group(P<0.05),and NF-κB was higher in the hypoxia group than the normoxia group(P<0.05).The normoxia group could inhibit autophagy proteins LC3B and Beclin1 by activating NF-κB through overexpression of OPN in PASMCs,and the addition of NF-κB inhibitor could promote the expression of autophagy proteins in PASMCs(P<0.05).Interfering with the expression of OPN in PASMCs in the hypoxia group could further promote the expression of autophagy proteins and autophagic vesicles(P<0.05).The inhibition of OPN and NF-κB expression in PASMCs could block the cell transition from G0/G1 phase to S phase and attenuate the proliferation induced by increased OPN(P<0.05).Conclusion Trans-targeted OPN activates the IKBα/NF-κB signaling pathway.The activated IKBα/NF-κB signaling pathway affects the autophagy,proliferation,and cell cycle of PASMCs in hypoxic environment.

关 键 词：骨桥蛋白 IKBα/NF-κB通路 自噬 低氧 肺动脉平滑肌细胞 增殖 

分 类 号：R818[医药卫生—放射医学]