基于网络药理学和实验验证：β-谷甾醇通过磷酸化MAPK3减轻顺铂诱导的听力损失

文献信息

中文摘要

目的： 本研究旨在探讨熟地黄及其生物活性成分β-谷甾醇在预防顺铂诱导的听力损失（CIHL）中的保护作用及其分子机制。
方法： 采用网络药理学和生物信息学分析鉴定熟地黄的药理活性成分并预测其潜在分子靶点。在体外和体内评估β-谷甾醇对毛细胞损伤、凋亡、活性氧（ROS）水平及靶蛋白表达的影响。
结果： 熟地黄的主要活性成分为β-谷甾醇和豆甾醇。它们与CIHL共有50个潜在靶点，其中MAPK3、SRC和ESR1位列前三。分子对接结合动力学模拟表明β-谷甾醇与MAPK3之间具有强结合并形成稳定复合物。在体外，β-谷甾醇减轻顺铂诱导的毛细胞死亡，降低ROS水平，并恢复磷酸化MAPK3。在体内，它改善了听力，减少了毛细胞丢失，并部分恢复了顺铂处理小鼠耳蜗中的磷酸化MAPK3。
结论： 熟地黄及其活性成分β-谷甾醇通过恢复MAPK3磷酸化信号通路保护耳蜗毛细胞免受顺铂诱导的损伤，为CIHL治疗提供了新的分子靶点和策略。

英文摘要

PURPOSE: This research aimed to investigate the protective role as well as molecular mechanisms of Rehmanniae Radix Praeparata and its bioactive compound β-sitosterol in preventing CIHL.
METHODS: Network pharmacology and bioinformatic analyses were used to identify the pharmacologically active constituents of Rehmanniae Radix Praeparata and predict their potential molecular targets. The influence of β-sitosterol on hair cell damage, apoptosis, reactive oxygen species (ROS) levels, and target protein expression were evaluated in vitro as well as in vivo.
RESULTS: The primary active constituents of Rehmanniae Radix Praeparata were β-sitosterol and stigmasterol. They shared 50 potential targets with CIHL, with MAPK3, SRC, and ESR1 as the top three. Molecular docking combined with dynamics simulations demonstrated strong binding and stable complex formation between β-sitosterol and MAPK3. In vitro, β-sitosterol mitigated cisplatin-induced hair cell death, reduced ROS levels, and restored phosphorylated MAPK3. In vivo, it enhanced hearing, decreased hair cell loss, and partially restored phosphorylated MAPK3 in the cochleae of cisplatin-treated mice.
CONCLUSION: Rehmanniae Radix Praeparata and its active component β-sitosterol protected cochlear hair cells from cisplatin-induced damage by restoring MAPK3 phosphorylation signaling pathway, providing new molecular targets and strategies for CIHL treatment.