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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200μg/m L)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、p38M APK蛋白及磷酸化程度。结果与对照组比较,100和200μg/m L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、200μg/m L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M IN6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。
Objective To investigate the effects of cigarette smoke extract (CSE) on the morphology and viability of pancreatic island MIN6 cells and the expression of ERK, JNK and p38MAPK in intracellular mitogen - activated protein kinase (MAPKs) signal transduction pathway. Methods MIN6 cells were cultured in vitro until the logarithmic growth phase. The experimental groups with different concentrations of CSE (20, 50, 100 and 200 μg / mL) were established. At the same time, the control group was given morphological changes of MIN6 cells after 48 h The inhibitory rate of CSE on the proliferation of MIN6 cells was detected by CCK-8 assay. Meanwhile, the protein and phosphorylation of ERK, JNK and p38M in the cells were detected by Western blotting (WB). Results Compared with the control group, the morphological changes of the cells under the doses of 100 and 200 μg / m L were obvious abnormalities, the nuclei were pyknotic and the spindle shape was changed. Compared with the control group, the viability of MIN6 cells decreased, The viability of cells in 200μg / mL group was (97.32 ± 2.67)%, (94.67 ± 5.33)%, (87.71 ± 12.3)% and (74.23 ± 25.8)%, respectively. Compared with the control group, the expressions of ERK, JNK and p38MAPK However, the phosphorylation of p-JNK and p-p38MAPK were significantly increased with the increase of CSE dose (P <0.05 compared with the control group), and showed a dose-response relationship. Conclusion CSE can induce abnormal morphology and inhibit the proliferation of M IN6 cells, which may be related to the activation of JNK and p38 MAPK signaling pathway.