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）激活是肝纤维化进程中的关键事件。在这项研究中，首先通过生物信息学分析预测海伦纳林的靶标，然后通过各种实验验证该预测。HSC-T6细胞被白介素-1β（IL-1β）激活，然后用海伦素处理。此外，用miR-200a模拟物或抑制剂转染了HSC-T6细胞，并研究了海伦素对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活化，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Helenalin的靶标可能是miR-200a介导的PI3K / Akt和NF-κB途径。此外，实验表明，Helenalin给药可导致HSC-T6细胞失活，这可通过抑制细胞增殖，α-SMA表达和胶原蛋白生成来证明。机制研究表明，海蓝素通过恢复MMP / TIMP的平衡来减少胶原蛋白的积累。此外，海伦纳林通过抑制PI3K / Akt途径显着抑制HSC活化，并通过阻断NF-κB信号转导减轻炎症反应。进一步的研究表明海伦纳林上调了miR-200a的表达，从而导致PI3K / Akt和NF-κB信号通路的抑制。总之，Helenalin通过抑制miR-200a介导的PI3K / Akt和NF-κB途径来抑制HSC活化，并且可能被开发为治疗肝纤维化的潜在药物。