Krupple样因子6及诱导型一氧化氮合酶在铜绿假单胞菌上清液感染RAW264.7凋亡过程中的表达
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摘要
目的检测Krupple样因子6 (Krupple-like factor 6,KLF6)及诱导型一氧化氮合酶(inducible nitric oxide synthase,i NOS)在铜绿假单胞菌(pseudomonas aeruginosa,PA)上清液感染的小鼠单核巨噬细胞白血病细胞(RAW264. 7)中的表达水平,探讨KLF6及i NOS在PA上清液感染RAW264. 7凋亡过程中的作用。方法 MTT法检测PA上清液对RAW264. 7增殖的影响;流式细胞术检测凋亡率; Hoechst 33342染色检测RAW264. 7细胞核形态变化; Western blot检测PA上清液对KLF6、i NOS表达的影响。结果 PA上清液抑制RAW264. 7的增殖,且其抑制作用呈浓度和时间依赖性,PA上清液能够明显增加RAW264. 7的凋亡比例,促进KLF6、i NOS的表达。结论 PA能够抑制RAW264. 7的增殖,其可能通过增加KLF6及i NOS的表达,从而诱导细胞凋亡。
Aim To detect the expression of Krupplelike factor 6( KLF6) and inducible nitric oxide synthase( i NOS) in mouse mononuclear macrophage leukemia cells infected with pseudomonas aeruginosa( PA) supernatant,and to investigate the role of KLF6 protein and i NOS protein in the apoptosis of RAW264. 7 infected with PA supernatant. Methods The effect of PA supernatant on the proliferation of RAW264. 7 was detected by MTT assay. The apoptotic rate was detected by flow cytometry. The morphological changes of RAW264. 7 were detected by Hoechst33342 staining. The effect of PA supernatant on KLF6and i NOS protein expression was detected by Western blot. Results PA supernatant inhibited the proliferation of RAW264. 7 in a concentration-and time-dependent manner. PA supernatant significantly increased the percentage of RAW264. 7 apoptosis and promoted the expression of KLF6 and i NOS. Conclusions PA can inhibit the proliferation of RAW264. 7,which may induce apoptosis through KLF6 and i NOS protein expression.
Aim To detect the expression of Krupplelike factor 6( KLF6) and inducible nitric oxide synthase( i NOS) in mouse mononuclear macrophage leukemia cells infected with pseudomonas aeruginosa( PA) supernatant,and to investigate the role of KLF6 protein and i NOS protein in the apoptosis of RAW264. 7 infected with PA supernatant. Methods The effect of PA supernatant on the proliferation of RAW264. 7 was detected by MTT assay. The apoptotic rate was detected by flow cytometry. The morphological changes of RAW264. 7 were detected by Hoechst33342 staining. The effect of PA supernatant on KLF6and i NOS protein expression was detected by Western blot. Results PA supernatant inhibited the proliferation of RAW264. 7 in a concentration-and time-dependent manner. PA supernatant significantly increased the percentage of RAW264. 7 apoptosis and promoted the expression of KLF6 and i NOS. Conclusions PA can inhibit the proliferation of RAW264. 7,which may induce apoptosis through KLF6 and i NOS protein expression.
引文
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[2]Rodrigo-Troyano A,Suarez-Cuartin G,PeiróM,et al.Pseudomonas aeruginosa resistance patterns and clinical outcomes in hospitalized exacerbations of COPD[J].Respirology,2016,21(7):1235-42.
[3]Jacome P,Alves L R,Jácome-Júnior A T,et al.Detection of blaSPM-1,blaKPC,blaTEM and blaCTX-M genes in isolates of Pseudomonas aeruginosa,Acinetobacter spp.and Klebsiella spp.from cancer patients with healthcare-associated infections[J].Med Microbiol,2016,65(7):658-65.
[4]Lovewell R R,Patankar Y R,Berwin B.Mechanisms of phagocytosis and host clearance of Pseudomonas aeruginosa[J].Am JPhysiol Lung Cell Mol Physiol,2014,306(7):L591-603.
[5]Andreoli V,Gehrau R C,Bocco J L.Biology of Kruppel-like factor6 transcriptional regulator in cell life and death[J].IUBMB Life,2010,62(12):896-905.
[6]Lee M,Rey K,Besler K,et al.Immunobiology of nitric oxide and regulation of inducible nitric oxide synthase[J].Results Probl Cell Differ,2017,62:181-207.
[7]Fite A,Abou-Samra A B,Seyoum B.Macrophages inhibit insulin signalling in adipocytes:role of inducible nitric oxide synthase and nitric oxide[J].Can J Diabetes,2015,39(1):36-43.
[8]喻志芳,韩彬,柴文戍.Krupple样因子6调控诱导型一氧化氮合酶介导铜绿假单胞菌感染小鼠肺组织细胞凋亡的研究[J].中国全科医学,2016,19(20):2435-9.[8]Yu Z F,Han B,Chai W S.KLF6 and i NOS regulated apoptosis during Pseudomonas Aeruginosa lung infection in mice[J].Chin Generat Practice,2016,19(20):2435-9.
[9]孟子卜,柴文戍.铜绿假单胞菌上清液诱导J774细胞凋亡过程中Fas蛋白的表达[J].中国药理学通报,2012,28(1):147.[9]Meng Z B,Chai W S.Expression of Fas protein during the cellfree supernatant from pseudomonas aeruginosa-induced apoptosis of J774 cells[J].Chin Pharmacol Bull,2012,28(1):147.
[10]Li T,Huang X,Yuan Z,Wang L,et al.Pyocyanin induces NK92cell apoptosis via mitochondrial damage and elevated intracellular Ca2+[J/OL].Innate Immun,2019,25(1):3-12.
[11]Kaminski A,Gupta K H,Goldufsky J W,et al.Pseudomonas aeruginosa ExoS induces intrinsic apoptosis in target host cells in a manner that is dependent on its GAP domain activity[J].Sci Rep,2018,8(1):14047.
[12]Mgbemena V,Segovia J,Chang T H,et al.KLF6 and i NOS regulates apoptosis during respiratory syncytial virus infection[J].Cell Immunol,2013,283(1-2):1-7.
[13]Kamijo R,Harada H,Matsuyama T,et al.Requirement for transcription factor IRF-1 in NO synthase induction in macrophages[J].Science,1994,263(5 153):1612-5
[14]王春波,李金莲,张正洋,等.扇贝多肽对UVB照射Ha CaT细胞后NO释放及热休克蛋白70表达的影响[J].中国药理学通报,2011,27(3):334-7.[14]Wang C B,Li J L,Zhang Z Y,et al.,Effect of polypeptide from Chlamys farreri on UVB-induced NO release and HSP70 expression in Ha CaT cells[J].Chin Pharmacol Bull,2011,27(3):334-7.
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