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汪秋实, 曹烨, 闫丽, 韩翱瀚, 覃玉, 吕淑荣, 向全永. 香烟烟雾提取物对MIN6细胞JNK及p38MAPK表达影响[J]. 中国公共卫生, 2016, 32(12): 1689-1692. DOI: 10.11847/zgggws2016-32-12-21
引用本文: 汪秋实, 曹烨, 闫丽, 韩翱瀚, 覃玉, 吕淑荣, 向全永. 香烟烟雾提取物对MIN6细胞JNK及p38MAPK表达影响[J]. 中国公共卫生, 2016, 32(12): 1689-1692. DOI: 10.11847/zgggws2016-32-12-21
WANG Qiu-shi, CAO Ye, YAN Li.et al, . Effect of cigarette smoke extract on JNK, p38MAPK expressions in MIN6 cells[J]. Chinese Journal of Public Health, 2016, 32(12): 1689-1692. DOI: 10.11847/zgggws2016-32-12-21
Citation: WANG Qiu-shi, CAO Ye, YAN Li.et al, . Effect of cigarette smoke extract on JNK, p38MAPK expressions in MIN6 cells[J]. Chinese Journal of Public Health, 2016, 32(12): 1689-1692. DOI: 10.11847/zgggws2016-32-12-21

香烟烟雾提取物对MIN6细胞JNK及p38MAPK表达影响

Effect of cigarette smoke extract on JNK, p38MAPK expressions in MIN6 cells

  • 摘要: 目的 探讨体外香烟烟雾提取物(CSE)对胰岛MIN6细胞形态及活力的影响,及对胞内丝裂原活化蛋白激酶(MAPKs)信号转导通路ERK、JNK、p38MAPK表达的影响。方法 体外培养MIN6细胞至生长对数期后,设不同浓度CSE(20、50、100和200 μg/mL)实验组,同时设对照组,48 h后,镜下观察MIN6细胞形态学改变,并应用CCK-8法检测CSE对MIN6细胞的增殖抑制率;同时应用蛋白印迹法(WB)检测细胞内ERK、JNK、p38MAPK蛋白及磷酸化程度。结果 与对照组比较,100和200 μg/mL剂量下的细胞形态出现明显异常,胞核固缩,细胞呈梭形样改变;与对照组比较,MIN6细胞活力出现下降,20、50、100、200 μg/mL各组细胞活力分别为(97.32±2.67)%、(94.67±5.33)%、(87.71±12.3)%和(74.23±25.8)%;与对照组相比,ERK、JNK、p38MAPK蛋白表达变化不大,但随着CSE剂量的增加,p-JNK和p-p38MAPK磷酸化程度明显增加(与对照组相比,P<0.05),并呈一定的剂量-效应关系。结论 CSE能够导致MIN6细胞形态异常、抑制其细胞增殖活力,其机制可能与JNK和p38MAPK信号通路的激活相关。

     

    Abstract: Objective To investigate the effect of cigarette smoke extract (CSE) on expressions of c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MAPK) in MIN6 cells.Methods MIN6 cells in logarithmic growth period were cultured in vitro and then treated with different concentrations of CSE (20,50,100,200 μg/mL) and a vehicle (control) group was also established.After 48 hours' treatment of CSE,the change in morphology of the cells was observed with inverted microscope and cholecystokinin (CCK-8) was used to assess CSE-mediated anti-proliferative effect on MIN6 cells.Meanwhile,Western blot analysis was applied to determine cellular protein expressions of extracellular-signal-regulated kinase (ERK),JNK,p38MAPK,and corresponding phosphorylation level.Results Compared with the control group,obvious abnormal morphology was found in the cells under the treatment of 100 μg/mL and 200 μg/mL CSE,including nuclear condensation and spindle like change.Compared with the control group,CSE at different dosages significantly inhibited the viability of MIN6 cells,with the cell viabilities of 97.32±2.67%,94.67±5.33%,87.71±12% (all P<0.05),and 74.23±25.80%(P<0.01) for the cells treated with 20,50,100,and 200 μg/mL CSE,respectively.In addition,CSE effectively activated the JNK and p38MAPK signaling pathway.With the increment of CSE concentration,the phosphorylation level significantly increased when compared to that of the control group (P<0.05).Conclusion Cigarette smoke extract could decrease the cell viability of MIN6 cells and the effect may due to the increased expressions of p-JNK and p-p38MAPK.

     

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