[1]于晓晴,钟根深,熊熙文,等.三磷腺苷合酶抑制因子1基因敲除对小鼠巨噬细胞线粒体功能和三磷腺苷水平的影响[J].新乡医学院学报,2019,36(6):501-505.[doi:10.7683/xxyxyxb.2019.06.001]
　YU Xiao-qing,ZHONG Gen-shen,XIONG Xi-wen,et al.Effects of adenosine triphosphate synthase inhibitor 1 gene knockout on the levels of adenosine triphosphate and mitochondrial function in mouse macrophages[J].Journal of Xinxiang Medical University,2019,36(6):501-505.[doi:10.7683/xxyxyxb.2019.06.001]
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三磷腺苷合酶抑制因子1基因敲除对小鼠巨噬细胞线粒体功能和三磷腺苷水平的影响
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
36
期数:
2019年6
页码:
501-505
栏目:
基础研究
出版日期:
2019-06-05

文章信息/Info

Title:
Effects of adenosine triphosphate synthase inhibitor 1 gene knockout on the levels of adenosine triphosphate and mitochondrial function in mouse macrophages
作者:
于晓晴1; 2; 钟根深1; 2; 熊熙文3; 梁银明1; 2; 王　辉1; 2; 章小英4; 叶建平4
（1.新乡医学院医学检验学院，河南　新乡　453003；2.河南省分子诊断与医学检验技术协同创新中心，河南　新乡　453003；3.新乡医学院医学法医学院，河南　新乡　453003；4.上海交通大学第六人民医院中心实验室，上海　201306）
Author(s):
YU Xiao-qing1; 2; ZHONG Gen-shen1; 2; XIONG Xi-wen3; LIANG Yin-ming1; 2; WANG Hui1; 2; ZHANG Xiao-ying4; YE Jian-ping4
（1.School of Laboratory Medicine，Xinxiang Medical University，Xinxiang 453003，Henan Province，China；2.Henan Province Collaborative Innovation Center of Molecular Diagnosis and Laboratorial Medicine，Xinxiang 453003，Henan Province，China；3.School of Forensic Medicine，Xinxiang Medical University，Xinxiang 453003，Henan Province,China；4.Central Laboratory,the Sixth People′s Hospital Affiliated to Shanghai Jiaotong University，Shanghai 201306，China）
关键词:
ATP合酶抑制因子1; 巨噬细胞; 线粒体; 三羧酸循环; 氧耗量
Keywords:
adenosine triphosphate synthase inhibitor 1; macrophage; mitochondria; tricarboxylic acid cycle; oxygen consumption
分类号:
R589.2
DOI:
10.7683/xxyxyxb.2019.06.001
文献标志码:
A
摘要:
目的　探讨三磷腺苷合酶抑制因子1（ATPIF1）基因敲除对小鼠巨噬细胞内三磷腺苷（ATP）水平及线粒体功能的影响。方法　选取5只野生型C57BL/6小鼠为WT组，5只ATPIF1基因敲除小鼠为KO组。处死小鼠后，提取小鼠骨髓细胞进行培养，并诱导其分化成熟为骨髓来源巨噬细胞（BMDM）。在未加脂多糖（LPS）的基础状态及LPS刺激2、4 h 后收集细胞，应用ATP检测试剂盒检测细胞内ATP水平，应用细胞能量代谢分析仪检测细胞氧消耗率，采用实时荧光定量聚合酶链反应检测细胞内三羧酸循环中关键酶丙酮酸激酶（PKm）、异柠檬酸脱氢酶（IDH）、柠檬酸合酶 （CS）、α-酮戊二酸脱氢酶（OGDC）mRNA相对表达量。结果　与WT组比较，在基础状态下KO组小鼠BMDM 内ATP水平、基础氧耗量、用于ATP合成的氧耗量、最大氧耗量及PKm、IDH、CS、OGDC mRNA相对表达量显著增加（P<0.05，P<0.01）；LPS刺激2、4 h后，KO组小鼠BMDM 内ATP水平、基础氧耗量、用于ATP合成的氧耗量和最大氧耗量及CS、OGDC mRNA相对表达量均显著增加（P<0.05）。结论　ATPIF1基因敲除可增加小鼠BMDM内ATP水平，并提高三羧酸循环相关酶mRNA表达和线粒体氧化磷酸化水平。
Abstract:
Objective　To investigate the effect of adenosine triphosphate synthase inhibitor 1 （ATPIF1） gene knockout on adenosine triphosphate （ATP） level and mitochondrial function of mouse macrophages.Methods　Five wild type C57BL/6 mice were selected as WT group，and five ATPIF1 gene knockout mice were selected as KO group.After mice were sacrificed，bone marrow cells were cultured and induced to differentiate into bone marrow-derived macrophages （BMDM）.Then cells were collected at the basic state without lipopolysaccharide （LPS） and at 2，4 hours after LPS stimulation，then ATP level in the BMDM was detected by ATP assay kit；cell oxygen consumption rate was measured by cell energy metabolism analyzer.The relative mRNA expression levels of key enzymes in the cell tricarboxylic acid cycle，including pyruvate kinase （PKm），isocitrate dehydrogenase （IDH），citrate synthase （CS） and α-ketoglutarate dehydrogenase （OGDC） was detected by real-time fluorescence quantitative polymerase chain reaction.Results　Compared with the WT group，in the basic state，the ATP level，basic oxygen consumption，oxygen consumption for ATP synthesis,maximum oxygen consumption，and the relative mRNA expressions of key enzymes PKm,IDH,CS and OGDC were significantly increased （P<0.05，P<0.01）.At 2，4 hours after LPS stimulation，the ATP level，the basic oxygen consumption，oxygen consumption for ATP synthesis，maximum oxygen consumption and the mRNA expression levels of CS and OGDC of tricarboxylic acid cycle genes in the KO group were significantly higher compared with those in the WT group（P<0.05）.Conclusion　ATPIF1 gene knockout increased the ATP level in the BMDM of mice，and increased the expression of mRNA of key enzymes of the tricarboxylic acid cycle and increased the level of mitochondrial oxidative phosphorylation.

参考文献/References:

［1］　BONORA M，PATERGNANI S，RIMESSI A，et al.ATP synthesis and storage［J］.Purinergic Signal，2012，8（3）：343-357.
［2］　HARA K Y，KONDO A.ATP regulation in bioproduction［J］.Microb Cell Fact，2015，14：198.
［3］　GARCIA-BERMUDEZ J，CUEZVA J M.The ATPase inhibitory factor 1 （IF1）：a master regulator of energy metabolism and of cell survival［J］.Biochim Biophys Acta，2016，1857（8）：1167-1182.
［4］　BASON J V，MONTGOMERY M G，LESLIE A G，et al.Pathway of binding of the intrinsically disordered mitochondrial inhibitor protein to F1-ATPase［J］.Proc Natl Acad Sci USA，2014，111（31）：11305-11310.
［5］　SANTACATTERINA F，SANCHEZ-CENIZO L，FORMENTINI L，et al.Down-regulation of oxidative phosphorylation in the liver by expression of the ATPase inhibitory factor 1 induces a tumor-promoter metabolic state［J］.Oncotarget，2016，7（1）：490-508.
［6］　FORMENTINI L，SANCHEZ-ARAGO M，SANCHEZ-CENIZO L，et al.The mitochondrial ATPase inhibitory factor 1 triggers a ROS-mediated retrograde prosurvival and proliferative response［J］.Mol Cell，2012，45（6）：731-742.
［7］　FORMENTINI L，PEREIRA M P，SANCHEZ-CENIZO L，et al.In vivo inhibition of the mitochondrial H+-ATP synthase in neurons promotes metabolic preconditioning［J］.EMBO J，2014，33（7）：762-778.
［8］　CHIN R M，FU X，PAI M Y，et al.The metabolite alpha-ketoglutarate extends lifespan by inhibiting ATP synthase and TOR［J］.Nature，2014，510（7505）：397-401.
［9］　关丫丫，梁银明，王辉，等.三磷腺苷合酶抑制因子1基因敲除对小鼠成纤维细胞中三磷腺苷水平及脂肪细胞分化的影响［J］.新乡医学院学报，2018，35（8）：658-661.
［10］　ESPARZA-MOLTO P B，NUEVO-TAPIOLES C，CUEZVA J M.Regulation of the H+-ATP synthase by IF1：a role in mitohormesis［J］.Cell Mol Life Sci，2017，74（12）：2151-2166.
［11］　GARCIA-AGUILAR A，CUEZVA J M.A review of the inhibition of the mitochondrial ATP synthase by IF1 in vivo：reprogramming energy metabolism and inducing mitohormesis［J］.Front Physiol，2018，9：1322.
［12］　GARCIA-BERMUDEZ J，SANCHEZ-ARAGO M，SOLDEVILLA B，et al.PKA phosphorylates the ATPase inhibitory factor 1 and inactivates its capacity to bind and inhibit the mitochondrial H+-ATP synthase［J］.Cell Rep，2015，12（12）：2143-2155.
［13］　SANCHEZ-ARAGO M，FORMENTINI L，MARTINEZ-REYES I，et al.Expression，regulation and clinical relevance of the ATPase inhibitory factor 1 in human cancers［J］.Oncogenesis，2013，2：e46.
［14］　SANCHEZ-ARAGO M，GARCIA-BERMUDEZ J，MARTINEZ-REYES I，et al.Degradation of IF1 controls energy metabolism during osteogenic differentiation of stem cells［J］.EMBO Reports，2013，14（7）：638-644.
［15］　SANTACATTERINA F，SNCHEZ-CENIZO L，FORMENTINI L，et al.Down-regulation of oxidative phosphorylation in the liver by expression of the ATPase inhibitory factor 1 induces a tumor-promoter metabolic state［J］.Oncotarget，2016，7（1）：490-508.
［16］　ANDREAS K，BORIS M，LIMBERG S H，et al."Host tissue damage" signal ATP promotes non-directional migration and negatively regulates toll-like receptor signaling in human monocytes［J］.J Biol Chem，2005，280（37）：32459-32467.
［17］　HAM J，REES D A.The adenosine a2b receptor：its role in inflammation［J］.Endocr Metab Immune Disord Drug Targets，2008，8（4）：244-254.
［18］　VITIELLO L，GORINI S，ROSANO G，et al.Immunoregulation through extracellular nucleotides［J］.Blood，2012，120（3）：511-518.
［19］　LEE J H，ZHANG Y，ZHAO Z，et al.Intracellular ATP in balance of pro- and anti-inflammatory cytokines in adipose tissue with and without tissue expansion［J］.Int J Obes （Lond），2017，41（4）：645-651.
［20］　CHANG Y P，KA S M，HSU W H，et al.Resveratrol inhibits NLRP3 inflammasome activation by preserving mitochondrial integrity and augmenting autophagy［J］.J Cell Physiol，2015，230（7）：1567-1579.

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更新日期/Last Update: 2019-06-05