晚期氧化蛋白产物调节人肾小管上皮细胞自噬的研究  

Advanced oxidation protein products modulate autophagy of renal tubule epithelial cells

作　　者：章俊[1] 向晓红[1] 梁秀洁 束双双[1] 姜婷婷[1] 郭婷婷[1] 汤珣[1] 

机构地区：[1]南方医科大学珠江医院肾内科,广州510280

出　　处：《重庆医学》2017年第25期3480-3483,共4页Chongqing medicine

基　　金：国家自然科学基金资助项目(81202842)

摘　　要：目的探讨晚期氧化蛋白产物(AOPPs)对人肾小管上皮细胞(HK-2)自噬的影响。方法 AOPPs刺激HK-2细胞,采用RT-qPCR和Western blot检测自噬相关蛋白LC3-Ⅱ/LC3-Ⅰ、Beclin1和p62的表达;Western blot检测p38MAPK通路的激活;加入p38 MAPK抑制剂(SB203580)与AOPPs共处理,观察自噬的改变,加入自噬诱导剂雷帕霉素与AOPPs共处理,并用RT-qPCR和Western blot检测细胞周期抑制蛋白p27的表达,用BCA法检测细胞总蛋白,观察细胞肥大的改变。结果AOPPs下调LC3-Ⅱ/LC3-Ⅰ、Beclin1,上调p62的表达,并激活p38 MAPK通路;与AOPPs单独处理组相比,SB203580与AOPPs共处理组LC3-Ⅱ/LC3-Ⅰ和Beclin升高而p62降低;雷帕霉素与AOPPs共处理组p27的表达和细胞总蛋白下调。结论AOPPs通过激活p38 MAPK通路抑制HK-2细胞自噬,自噬抑制参与HK-2细胞肥大。Objective To investigate the effect of advanced oxidation protein products （AOPPs） on human renal tubular epithelial cells（HK-2） autophagy. Methods HK-2 cells were stimulated with AOPPs. RT-qPCR and Western blot were used to determine the expression of autophagy related protein LC3-Ⅱ/LC3- Ⅰ , Beclinl and p62 ;Western blot was utilized to examine the activa- tion of p38 MAPK pathway. Then p38 MAPK inhibitor （SB203580） was added and co-processed with AOPPs. The change of auto- phagy was observed. Also, autophagy inducer rapamycin was added and co-processed with AOPPs. RT-qPCR and Western blot were used to detect the expression of cell cycle inhibitory protein p27. The cell total protein level was detected by the bicinchoninic acid （BCA） method. The hypertrophy change was observed. Results AOPPs down-regulated the expression of LC3- Ⅱ/LC3- Ⅰ and Be- clinl, up-regulated expression of p62 and activated p38 MAPK pathway;in comparison with the AOPPs alone treatment group,the expression of LC3-Ⅱ/LC3-Ⅰ and Beclin in the SB203580 co-processing group was increased,while p62 was decreased;the p27 ex- pression and ceils total protein in the sirolimus co-processing group were down-regulated. Conclusion AOPPs inhibits the autoph- agy of HK-2 cells by activating p38 MAPK pathway and the autophagy inhibition participates in HK-2 cell hypertrophy.

关 键 词：自噬 P38MAPK信号通路 晚期氧化蛋白产物 

分 类 号：R587.2[医药卫生—内分泌] R692.9[医药卫生—内科学]