School of Pharmaceutical Sciences, Sun Yat-Sen University, 132 East Outer Ring Road, Guangzhou 510006, China.
Key Laboratory of Preclinical Study for New Drugs of Gansu Province, School of Basic Medical Sciences, Lanzhou University, Lanzhou 730000, China.
Life Sci. 2019 Jul 15;229:200-209. doi: 10.1016/j.lfs.2019.04.037. Epub 2019 Apr 29.
The rapeseed protein derived peptide DHNNPQIR (named as RAP-8) has been previously reported to possess antioxidant activity and alleviate liver fibrosis. The purpose of the present study was to investigate the potential crucial pathways involved in ameliorating liver fibrosis of RAP-8.
Next-generation sequencing of messenger RNA (RNA-Seq) analysis of the fibrotic and RAP-8 treated mice was performed. Western blot, qPCR and flow cytometry detection analysis were conducted to measure cell cycle and oxidative stress in LX-2 cells and liver samples.
588 overlapped differentially expressed genes were obtained from a batch of genes RAP-8 altered. Gene Ontology enrichment analysis revealed that changes in the most significant modules were mainly enriched in cell division, nuclear division and mitotic cell cycle process, while alterations in Kyoto Encyclopedia of Genes and Genomes were mainly enriched in cell cycle. Thereafter, according to the co-expression network analysis, the regulations of three core genes (Cenpp, Cyp2c55, Serpinh1) were verified that might be targets for treating liver fibrosis. Furthermore, through experimental verification, we demonstrated that RAP-8 induced cell cycle arrest and prevents oxidation stress.
As a promising therapeutic candidate for hepatic fibrosis treating, RAP-8 exhibited anti-fibrotic effects via exerting cell cycle arrest and inhibiting oxidative stress.
先前有研究报道，油菜籽蛋白衍生肽 DHNNPQIR（命名为 RAP-8）具有抗氧化活性，并能减轻肝纤维化。本研究旨在探讨 RAP-8 改善肝纤维化的潜在关键途径。
对纤维化和 RAP-8 处理的小鼠进行信使 RNA（RNA-Seq）分析的下一代测序。通过 Western blot、qPCR 和流式细胞术检测分析，测量 LX-2 细胞和肝组织样本中的细胞周期和氧化应激。
从 RAP-8 改变的一批基因中获得了 588 个重叠差异表达基因。基因本体富集分析显示，最显著模块的变化主要富集在细胞分裂、核分裂和有丝分裂细胞周期过程中，而京都基因与基因组百科全书的改变主要富集在细胞周期中。此后，根据共表达网络分析，验证了三个核心基因（Cenpp、Cyp2c55、Serpinh1）的调节作用，这三个基因可能是治疗肝纤维化的靶点。此外，通过实验验证，我们证明 RAP-8 诱导细胞周期停滞并防止氧化应激。
作为一种有前途的肝纤维化治疗候选药物，RAP-8 通过发挥细胞周期停滞和抑制氧化应激作用发挥抗纤维化作用。
