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作 者:叶萌[1] 张夏男[1] 李跃纲[2] 贾效伟[1] 刘秉慈[1]
机构地区:[1]中国疾病预防控制中心职业卫生与中毒控制所化学污染物与健康重点实验室,北京100050 [2]北京市疾病预防控制中心营养与食品卫生所
出 处:《卫生研究》2014年第2期198-202,共5页Journal of Hygiene Research
基 金:国家自然科学基金(No.30972449;30671747)
摘 要:目的探讨铜蓝蛋白(Cp)在石英诱导的人胚肺成纤维细胞(HELF)中JNK/ERK/AP-1信号通路改变中的作用。方法分别在100μg/ml石英作用前1 h、同时和作用后1h加入30μg/ml Cp,同时设立单独用石英和单独用Cp处理以及不做任何处理的细胞组,刺激细胞24 h后,用噻唑蓝(MTT)比色法观察Cp对石英诱导的HELF细胞增殖的影响。用100μg/ml石英分别作用于HELF细胞、转染JNK显性失活突变体质粒的HELF细胞(DN-JNK)和转染ERK显性失活突变体质粒的HELF细胞(DN-ERK)24 h;用100μg/ml石英作用1 h后,加入30μg/ml Cp作用24 h;同时设立不做任何处理的对照组。用MTT法检测分别抑制JNK和ERK后对细胞增殖的影响;用蛋白免疫印迹(Western blotting)实验检测JNK、ERK、c-Jun、c-Fos及其磷酸化水平的改变。结果石英作用1 h后加入Cp,对石英诱导的细胞增殖有促进作用,后续的实验选择这种作用模式。Cp可以明显增加JNK、ERK蛋白及磷酸化ERK(pERK)、磷酸化JNK(p-JNK)、磷酸化c-Jun(p-c-Jun)和磷酸化c-Fos(p-c-Fos)蛋白水平。抑制JNK蛋白活性后,Cp诱导的p-JNK、p-c-Jun和p-c-Fos蛋白水平的增加被抑制,Cp、石英以及Cp和石英共同诱导的细胞增殖加快被抑制;抑制ERK蛋白后,Cp诱导的p-ERK和p-c-Fos的增加被抑制,Cp、石英以及Cp和石英共同诱导的细胞增殖加快被抑制。结论 Cp通过JNK/c-Jun和c-Fos及ERK/c-Fos通路进一步增强石英诱导的促细胞增殖作用。Objective To investigate the roles of ceruloplasmin (Cp) in JNK/ERK/ AP-1 cell signaling pathway change in human embryonic lung fibroblasts (HELFs) induced by silica. Methods Cp stimulated HELFs in different time points (before I h, accompanied with or after 1 h of silica-adding). HELFs were divided into these groups: control group, silica(100 μg/ml for 24 h) group, Cp (30μg/ml for 24 h) group and silica plus Cp (100μg/ml silica plus 30 μg/ml Cp) group. DN-JNK ceils and DN-ERK cells (cells were transfected with dominant negative mutant plasmid) contained thesegroups: control group, silica group, silica plus Cp group. MTT assay was used to detect the effects of Cp on silica-induced cell proliferation. Western blot assay was performed to detect the levels of JNK, ERK, c-Jun, c-Fos and their phosphorylated levels. Results Cp promoted cell proliferation induced by silica when silica stimulated HELFs 1 h then adding to Cp. Cp significantly increased silica-induced the high levels of JNK, ERK and phosphorylated JNK (p-JNK) , p-ERK, p-c-Jun and p-c-fos protein. After inhibition of JNK or ERK, silica-and-Cp-induced cell proliferation was markedly decreased. When suppressing JNK protein, the increased levels of p-JNK, p-c-Jun and p-c-fos protein was not observed. The high levels of p-ERK, p-c-Jun and p-c-fos protein were decreased when inhibiting ERK protein. Conclusion Cp could further strengthen silica-induced cell proliferation by JNK/c-Jun/c-Fos and ERK/c-Jun cell signaling pathway.
关 键 词:石英 铜蓝蛋白 丝裂素活化蛋白激酶 活化蛋白 细胞增殖
分 类 号:R135.2[医药卫生—劳动卫生] Q518.4[医药卫生—公共卫生与预防医学]
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