哺乳动物细胞中调控AQP9磷酸化的蛋白因子及对砷摄入影响的研究

发布时间：2018-04-23 23:31

本文选题：AQP + 磷酸化　；参考：《中国药学杂志》2015年04期

【摘要】：目的探讨蛋白激酶p38,蛋白激酶C对几种哺乳动物细胞中水通道蛋白9磷酸化的调控作用,以及这些蛋白因子对细胞中砷摄入含量的影响。方法采用免疫印迹法和免疫共沉淀技术分别检测细胞株中p38,蛋白激酶C和水通道蛋白9表达水平及其磷酸化水平。电感藕合等离子体质谱法(ICP-MS)测定细胞内砷含量。结果在ECV-304,AsRE,Hep G2和L-02细胞株中,p38蛋白及其磷酸化表达水平在砷的刺激下随时间增加而有显著上升,相同条件下蛋白激酶C末见明显变化。当抑制细胞中p38活性,除了L-02细胞中水通道蛋白9的磷酸化水平受到抑制外,其余细胞中水通道蛋白9的蛋白含量及磷酸化水平都未见明显变化。砷含量也只在L-02细胞中看到有较显著的下降。结论水通道蛋白9磷酸化水平影响砷进入细胞的速度与含量,但不同的细胞中水通道蛋白9的磷酸化调控机制有所差异。p38蛋白激酶在正常肝细胞L-02中显示对水通道蛋白9磷酸化有一定的调控作用,蛋白激酶C则在本实验条件下未见入调控砷的入胞机制。
[Abstract]:Objective to investigate the effects of protein kinase p38 and protein kinase C on the phosphorylation of aquaporin 9 in several mammalian cells and the effect of these protein factors on arsenic uptake. Methods the expression and phosphorylation of p38 protein kinase C and aquaporin-9 were detected by Western blot and co-immunoprecipitation respectively. Inductively coupled plasma mass spectrometry (ICP-MS) was used to determine the content of arsenic in cells. Results the expression of p38 protein and its phosphorylation in ECV-304 AsREHep G2 and L-02 cells increased significantly with arsenic stimulation, and protein kinase C showed obvious changes at the end of the same condition. When the p38 activity was inhibited, the phosphorylation level of aquaporin 9 in L-02 cells was inhibited, but the protein content and phosphorylation level of aquaporin 9 in other cells did not change significantly. Arsenic content was also significantly decreased in L-02 cells. Conclusion the phosphorylation level of aquaporin 9 affects the rate and content of arsenic entering into cells. However, the phosphorylation mechanism of aquaporin 9 in different cells is different. P38 protein kinase has a certain role in regulating the phosphorylation of aquaporin 9 in normal hepatocytes L-02. Protein kinase C (PKC) did not show the mechanism of arsenic regulation in this experiment.
【作者单位】：杭州师范大学健康管理学院预防医学系;
【基金】：国家自然科学基金资助项目(81001227,30860241) 浙江省重点创新团队项目(2011R50021) 浙江省大学生科技创新活动计划(新苗人才计划)(2013R421040)
【分类号】：R965

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