琼玉膏延缓衰老作用的候选靶蛋白分析

Target protein candidates of hypothalamus in aging rats with intervention by Qiongyugao

采用iTRAQ技术研究琼玉膏干预衰老大鼠模型的下丘脑蛋白,寻找候选靶蛋白,探索琼玉膏延缓衰老的作用机制。结果表明,琼玉膏可以提高大鼠血清GSH-Px及肝脏SOD活力。iTRAQ技术鉴定到的总蛋白为3 522个,FDR<1%。通过数据分析,鉴定出琼玉膏组与模型组比较,具有295种差异蛋白;模型组与空白组比较,具有20种差异蛋白;其中与空白组相比,模型组中下调而琼玉膏组中上调的蛋白有14种。在琼玉膏组和模型组的295种差异蛋白中,差异倍数≥1.30的差异蛋白有40种,最高为1.47。查阅文献及基因功能检索,筛选出琼玉膏延缓衰老的候选靶蛋白12种:ST18,Ptprc,PSMB8,INPP4B,Shc3,Pik3r1,PIP5K1C,Nampt,Rasgrp2,Asah2,Pdpk1,Map2k7。Western blot法检测下丘脑炎症通路NF-κB蛋白,模型组(0.96)相对空白组(0.85)表达量升高;而琼玉膏组(0.89)相对模型组则下降。Q-PCR检测PIP5K1C,Ptprc等与炎症相关的候选靶蛋白,与空白组相比,模型组中mRNA相对表达量显著下降(P<0.01),而琼玉膏组中则显著升高(P<0.01),与蛋白质组学数据分析一致。琼玉膏可能通过调节下丘脑炎性反应而延缓衰老的发生。

To explore the associated proteins of the hypothalamus in aging rat models with intervention by Qiongyugao（ QYG） based on iTRAQ technology,find out the target protein candidates and investigate the mechanism of delaying aging for Qiongyugao. The results showed that Qiongyugao increased GSH-Px activity in serum and SOD activity in liver; the total protein count identified by iTRAQ was 3 522,FDR 1%. There were 20 kinds of differential proteins between the blank group and model group; there were 295 kinds of differential proteins between model group and QYG group,and 40 kinds of them had a difference multiple ≥1. 30（ the maximum value was 1. 47）. Compared with blank group,there were 14 kinds of proteins that were down-regulated in model group and up-regulated in QYG group. Combined with literature search and gene function search,12 kinds of target protein candidates were screened out ： ST18,Ptprc,PSMB8,INPP4 B,Shc3,Pik3r1,PIP5K1 C,Nampt,Rasgrp2,Asah2,Pdpk1,and Map2k7. The expression of nuclear factorkappa B（ NF-κB） in the hypothalamic inflammatory pathway was detected by Western blot and the results showed that its expression level in model group（ 0. 96） was higher than that in control group（ 0. 85）,while its expression level in QYG group（ 0. 89） was lower than that in model group. Q-PCR results showed that the relative mRNA expression levels of PIP5K1 C and Ptprc in model group were significantly lower than those in blank group（ P 0. 01）; while compared with the model group,the mRNA expression levels of PIP5K1 C and Ptprc in QYG group were significantly increased（ P 0. 01）. This result was consistent with proteomics data. QYG may delay aging by regulating hypothalamic inflammatory reaction