Guangxi Medical University, Nanning, China.
Guangxi Medical University, Nanning, China.
Biomed Pharmacother. 2019 Jul;115:108975. doi: 10.1016/j.biopha.2019.108975. Epub 2019 May 14.
In our previous works, we highlight nine candidates (Cathepsin D, Lamp1, Tpi1, Fgb, FVII, Mst4, CDK4, Hdgf and Glud1) that might be key proteins of combination therapy anti-fibrosis in rats. In this research, in order to verify the function of candidates, gene or protein expression of the nine candidates was verified by Western blot and RT-qPCR, and enzyme-linked immunosorbent assay in vitro and vivo. The expression of Fgb, Tpi1, CDK4, Mst4 and FVII significantly changed and matched with previous results after combination treating fibrosis rats or hepatic stellate cells (HSCs). These proteins may play crucial roles in anti-fibrosis. In particular, FVII take on pivotal role in combination therapy against liver fibrosis. The viability and cycle of HSCs was determined using CCK8 assay and Flow Cytometry, respectively. The results indicate that an overexpression of FVII could accelerate HSC proliferation and reduce the pharmacologic sensitivity. Combination therapy may inhibit HSC proliferation by blocking cell cycle in S phase. Although further studies are necessary to determine the precise protein functions, this research provides the possible molecular mechanisms and signaling pathways of combination therapy against liver fibrosis.
在我们之前的工作中，我们重点介绍了九种候选蛋白（组织蛋白酶 D、Lamp1、Tpi1、Fgb、FVII、Mst4、CDK4、Hdgf 和 Glud1），它们可能是大鼠联合抗纤维化治疗的关键蛋白。在本研究中，为了验证候选蛋白的功能，通过 Western blot 和 RT-qPCR 以及体外和体内酶联免疫吸附试验验证了这九种候选蛋白的基因或蛋白表达。在联合治疗纤维化大鼠或肝星状细胞（HSCs）后，Fgb、Tpi1、CDK4、Mst4 和 FVII 的表达显著变化，与之前的结果相匹配。这些蛋白可能在抗纤维化中发挥关键作用。特别是，FVII 在抗肝纤维化联合治疗中起着关键作用。通过 CCK8 测定法和流式细胞术分别测定 HSCs 的活力和细胞周期。结果表明，FVII 的过表达可加速 HSC 的增殖并降低药物敏感性。联合治疗可能通过阻断 S 期细胞周期来抑制 HSC 增殖。尽管还需要进一步的研究来确定确切的蛋白功能，但本研究为抗肝纤维化联合治疗提供了可能的分子机制和信号通路。
