Guangxi Medical University, Nanning 530021, China.
The First Affiliated Hospital of Guangxi University of Chinese Medicine, Nanning 530023, China.
Int Immunopharmacol. 2021 Mar;92:107208. doi: 10.1016/j.intimp.2020.107208. Epub 2021 Jan 11.
Hepatic stellate cell (HSC) activation is a crucial event in the progress of liver fibrosis. In this study, the target of helenalin was firstly predicted by bioinformatics analysis, and then the prediction was verified by various experiments. HSC-T6 cells were activated by interleukin-1 beta (IL-1β) and then treated with helenalin. Moreover, HSC-T6 cells were transfected with miR-200a mimic or inhibitor, and the effect of helenalin on the miR-200a-mediated PI3K/Akt and NF-κB signaling pathways was investigated. The bioinformatics analysis indicated that miR-200a might regulate the PI3K/Akt pathway, NF-κB activation, Bcl-2 family and Caspases, ultimately affecting cell survival and apoptosis. Interestingly, the molecular docking demonstrated that the target of helenalin might be miR-200a-mediated the PI3K/Akt and NF-κB pathways. Moreover, the experiments showed that helenalin administration led to the inactivation of HSC-T6 cells, as evidenced by the inhibition of cell proliferation, α-SMA expression and collagen production. The mechanism studies showed that helenalin reduced collagen accumulation by restoring the balance of MMPs/TIMPs. Moreover, helenalin markedly suppressed HSC activation by inhibiting the PI3K/Akt pathway and alleviated inflammatory response by blocking the NF-κB signal transduction. Further study indicated that helenalin up-regulated miR-200a expression, thus leading to the inhibition of the PI3K/Akt and NF-κB signaling pathways. In conclusion, helenalin inhibits HSC activation via inhibiting the miR-200a-mediated PI3K/Akt and NF-κB pathways, and it may be developed as a potential medicine for the treatment of liver fibrosis.
肝星状细胞（HSC）激活是肝纤维化进展的关键事件。本研究首先通过生物信息学分析预测了白屈菜红碱的靶标，然后通过各种实验验证了预测。用白细胞介素-1β（IL-1β）激活 HSC-T6 细胞，然后用白屈菜红碱处理。此外，用 miR-200a 模拟物或抑制剂转染 HSC-T6 细胞，研究白屈菜红碱对 miR-200a 介导的 PI3K/Akt 和 NF-κB 信号通路的影响。生物信息学分析表明，miR-200a 可能调节 PI3K/Akt 通路、NF-κB 激活、Bcl-2 家族和 Caspases，最终影响细胞存活和凋亡。有趣的是，分子对接表明白屈菜红碱的靶标可能是 miR-200a 介导的 PI3K/Akt 和 NF-κB 通路。此外，实验表明，白屈菜红碱给药导致 HSC-T6 细胞失活，表现为细胞增殖、α-SMA 表达和胶原产生受到抑制。机制研究表明，白屈菜红碱通过恢复 MMPs/TIMPs 的平衡减少胶原堆积。此外，白屈菜红碱通过抑制 PI3K/Akt 通路显著抑制 HSC 激活，并通过阻断 NF-κB 信号转导减轻炎症反应。进一步的研究表明，白屈菜红碱上调 miR-200a 表达，从而抑制 PI3K/Akt 和 NF-κB 信号通路。总之，白屈菜红碱通过抑制 miR-200a 介导的 PI3K/Akt 和 NF-κB 通路抑制 HSC 激活，可能被开发为治疗肝纤维化的潜在药物。
