一种简易的细胞烟草烟雾暴露装置的构建
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摘要
目的设计一种简易的细胞烟草烟雾暴露装置,并用其检测烟草烟雾暴露对人支气管上皮(HBE)细胞自噬的影响。方法2017年3月份起在广东药科大学广东省生物活性药物研究重点实验室利用transwell小室和六孔板构建烟草烟雾暴露装置;利用该装置对HBE细胞进行烟草烟雾处理;细胞免疫荧光和蛋白质免疫印迹法检测自噬蛋白LC3B的表达。结果成功制作简易装置用于细胞烟草烟雾暴露。利用该装置将HBE细胞烟草烟雾暴露后出现明显的细胞自噬。结论制作的细胞烟草烟雾暴露装置能用于细胞烟草烟雾暴露的研究,该装置具有制作简易、价格低廉等优点。
Objective To design a simple cell tobacco smoke exposure device and test the effect on the HBE cells. Methods The transwell and 6-wells plates had been used for constructing the tobacco smoke exposure device since March 2017, and the device was used for the tobacco and smoking of HBE cells, and the expression of autophagy protein LC3 B was tested by the immunocytochemistry and western immunoblotting. Results The tobacco smoke exposure device was successfully made, and the obvious autophagy occurred after using the device for tobacco smoke exposure. Conclusion The self-made tobacco smoke exposure device for the study of the cell tobacco smoking exposure has the advantages of simple formulation and low price.
Objective To design a simple cell tobacco smoke exposure device and test the effect on the HBE cells. Methods The transwell and 6-wells plates had been used for constructing the tobacco smoke exposure device since March 2017, and the device was used for the tobacco and smoking of HBE cells, and the expression of autophagy protein LC3 B was tested by the immunocytochemistry and western immunoblotting. Results The tobacco smoke exposure device was successfully made, and the obvious autophagy occurred after using the device for tobacco smoke exposure. Conclusion The self-made tobacco smoke exposure device for the study of the cell tobacco smoking exposure has the advantages of simple formulation and low price.
引文
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[4]Guo T,Chai X,Liu Q,et al.Downregulation of P16 promotes cigarette smoke extract-induced vascular smooth muscle cell proliferation via preventing G1/S phase transition[J].Experimental&Therapeutic Medicine,2017,14(1):214.
[5]Gu XM,Wang XG,Sun J,et al.The role of Hu R in mediating snail expression in human small airway epithelium induced by cigarette smoke extract.[J].Zhonghua jie he he hu xi za zhi=Zhonghua jiehe he huxi zazhi=Chinese journal of tuberculosis and respiratory diseases,2017,40(7):515.
[6]Xu H,Sun Q,Lu L,et al.Micro RNA-218 acts by repressing TNFR1-mediated activation of NF-κB,which is involved in MUC5AC hyper-production and inflammation in smoking-induced bronchiolitis of COPD[J].Toxicology Letters,2017,280:171-180.
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[8]Thorne D,Wilson J,Kumaravel TS,et al.Measurement of oxidative DNA damage induced by mainstream cigarette smoke in cultured NCI-H292 human pulmonary carcinoma cells[J].Mutation Research/Genetic Toxicology and Environmental Mutagenesis,2009,673(1):3-8.
[9]Phillips J,Kluss B,Richter A,et al.Exposure of bronchial epithelial cells to whole cigarette smoke:assessment of cellular responses[J].Altern Lab Anim,2005,33(3):239-248.
[10]Adamson J,Azzopardi D,Errington G,et al.Assessment of an in vitro,whole cigarette smoke exposure system:The Borgwaldt RM20S 8-syringe smoking machine[J].Chemistry Central Journal,2011,5(1):50.
[11]Azzopardi D,Haswell LE,Foss-Smith G,et al.Evaluation of an air-liquid interface cell culture model for studies on the inflammatory and cytotoxic responses to tobacco smoke aerosols[J].Toxicology in Vitro,2015,29(7):1720-1728.
[12]Bhayana S,Baisantry A,Kraemer TD,et al.Autophagy in kidney transplants of sirolimus treated recipients:[J].Journal of Nephropathology,2017,6(2):90.
[13]Mauthe M,Reggiori F.Using microbes as a key tool to unravel the mechanism of autophagyand the functions of the ATG pro teins[J].Microbial Cell,2016,4(1):1-5.
[14]Yin Z,Pascual C,Klionsky D.Autophagy:machinery and regulation[J].Microbial Cell,2016,3(12):588.
[15]Schneider JL,Miller AM,Woesner ME.Autophagy and Schizophrenia:A Closer Look at How Dysregulation of Neuronal Cell Homeostasis Influences the Pathogenesis of Schizophrenia[J].Einstein Journal of Biology&Medicine Ejbm,2016,31(1&2):34.
[16]Zhou JS,Zhao Y,Zhou HB,et al.Autophagy Plays an Essential Role in Cigarette Smoke-Induced Expression of MUC5ACin Airway Epithelium[J].American Journal of Physiology Lung Cellular&Molecular Physiology,2016,310(11):L1042-L1052.
[17]Yu F,Jun A,Saburo I,et al.Suppression of autophagy by extracellular vesicles promotes myofibroblast differentiation in COPD pathogenesis[J].Journal of Extracellular Vesicles,2015,4:28388.