shock proteins Mucins Cigarette smoke extract Activated prot

  本文关键词：热休克蛋白70在气道黏液高分泌中的作用，，由笔耕文化传播整理发布。

热休克蛋白70在气道黏液高分泌中的作用及机制

Role and mechanism of heat shock protein 70 in airway hypersecretion

[1] [2] [3] [4] [5]

Zhou Zhiyu, Zhang Ting, Zhou Xiangdong, Li Qi, Liu Chunyi.（Department of Respiratory Medicine, Second Affiliated Hospital, Chongqing University Medical Science, Chongqing 400010

重庆医科大学附属第二医院呼吸内科,400010

文章摘要：目的 探讨热休克蛋白70 （HSP70）在气道黏液高分泌中的作用及机制.方法 用8％香烟提取物（CSE）处理人气道A549细胞后,分别给予HSP70抗体阻断剂、c-Jun氨基末端激酶（JNK）特异性抑制剂SP600125进行干预.实验共分4组：空白对照组（无血清培养基培养）、CSE刺激组（加入8％ CSE培养24 h）、HSP70抗体组（HSP70单抗预处理30 min,再给予8％ CSE继续培养24h）、SP600125组（加入JNK特异性抑制剂SP600125 30 μmol/L预处理30 min,再给予8％ CSE继续培养24 h）.酶联免疫吸附（ELISA）法检测各组中MUC5AC蛋白的相对表达量,逆转录-PCR检测MUC5AC mRNA的相对表达量,Western印迹法检测HSP70水平,Western印迹法检测JNK及激活蛋白-1（主要为c-Jun）磷酸化水平.结果 CSE刺激组细胞MUC5AC蛋白、MUC5AC mRNA、p-JNK、c-Jun、p-c-Jun相对表达量（0.52±0.07、0.64±0.11、0.73±0.06、0.67±0.10、0.67±0.09）均显著高于空白对照组（0.26±0.10、0.28±0.06、0.30±0.05、0.30±0.08、0.36±0.08）、HSP70抗体组（0.30±0.12、0.29±0.09、0.34±0.06、0.47±0.19、0.39±0.13）和SP600125组（0.38±0.06、0.31±0.14、0.39±0.04、0.44±0.12、0.48±0.11）,HSP70 （0.75±0.09）也显著高于空白对照组（0.29±0.03）、HSP70抗体组（0.40±0.11）（均P＜0.05）.结论 HSP70可能通过JNK/激活蛋白-1信号通路上调气道A549细胞中MUC5AC的表达.

Abstr：Objective To explore the role and mechanism of heat shock protein 70 （HSP70） in airway hypersecretion.Methods After a stimulation of 8% cigarette smoke extract （CSE）,airway cells A549 were treated with HSP70 antibody and c-Jun N-terminal kinase （JNK） specific inhibitor SP600125 respectively.And the cells were divided into 4 groups of blank contrast （serum-free medium）,CSE stimulation （8% CSE for 24 h）,HSP70 antibody （30-min pre-treatment of HSP70 antibody and culturing in 8% CSE for 24 h） and SP600125 （30-min pre-treatment of SP600125 30 μmol/L and culturing in 8% CSE for 24 h）.The relative expression levels of MUC5AC protein in various groups were determined by enzymelinked immunosorbent assay （ELISA）.And the relative transcription level of MUC5AC mRNA was detected by reverse transcription-polymerase chain reaction （PCR） while the synthesized levels of HS70 as well as the phosphorylation levels of JNK and activated protein-1 （mostly c-Jun） were measured by Western blot.Results As compared with those in blank contrast group （0.26 ± 0.10,0.28 ± 0.06,0.30 ± 0.05,0.30±0.08,0.36±0.08）,HSP70 antibody group （0.30 ±0.12,0.29 ±0.09,0.34 ±0.06,0.47 ± 0.19,0.39 ±0.13） and SP600125 group （0.38 ±0.06,0.31 ±0.14,0.39 ±0.04,0.44 ±0.12,0.48 ± 0.11）,the relative expression levels of MUC5AC protein and mRNA,phosphorylation JNK （p-JNK）,c-Jun and p-c-Jun （0.52 ±0.07,0.64 ±0.11,0.73 ±0.06,0.67 ±0.10,0.67 ±0.09） significantly increased in CSE stimulation group （all P 〈 0.05）.And the synthesis levels of HSP70 （0.75 ± 0.09） in CSE stimulation group increased than blank contrast group （0.29 ± 0.03） and HSP70 antibody group （0.40 ± 0.11） （all P 〈 0.05）.Conclusion HSP70 may enhance the expression of MUC5AC in bronchial epithelial A549 cells via a

文章关键词：

Keyword：:HSP70 heat-shock proteins Mucins Cigarette smoke extract Activated protein

课题项目：国家自然科学基金（81370111）

作者信息：会员可见

  本文关键词：热休克蛋白70在气道黏液高分泌中的作用，由笔耕文化传播整理发布。