Abstract：
BACKGROUND: According to the Compendium of Materia Medica (Shizhen Li, Ming dynasty) and Welfare Pharmacy (Song dynasty), Psoraleae Fructus (PF), a traditional Chinese medicine (TCM) has a bitter taste and warm nature, which has the effect of treating spleen and kidney deficiency and skin disease. Although PF has been widely used since ancient times and has shown satisfactory efficacy in treating vitiligo, the active substances and the mechanism of PF in promoting melanogenesis remain unclear.
OBJECTIVE: To explore the active substances and action mechanisms of PF in promoting melanogenesis.
METHODS: Firstly, UPLC-UV-Q-TOF/MS was used to characterize the components in PF extract and identify the absorption components and metabolites of PF after oral administration at usual doses in rats. Secondly, the active substances and related targets and pathways were predicted by network pharmacology and molecular docking. Finally, pharmacodynamic and molecular biology experiments were used to verify the prediction results.
RESULTS: The experimental results showed that 15 compounds were identified in PF extract, and 44 compounds, consisting of 8 prototype components and 36 metabolites (including isomers) were identified in rats\' plasma. Promising action targets (MAPK1, MAPK8, MAPK14) and signaling pathways (MAPK signaling pathway) were screened and refined to elucidate the mechanism of PF against vitiligo based on network pharmacology. Bergaptol and xanthotol (the main metabolites of PF), psoralen (prototype drug), and PF extract significantly increased melanin production in zebrafish embryos. Furthermore, bergaptol could promote the pigmentation of zebrafish embryos more than psoralen and PF extract. Bergaptol significantly increased the protein expression levels of p-P38 and decreased ERK phosphorylation in B16F10 cells, which was also supported by the corresponding inhibitor/activator combination study. Moreover, bergaptol increased the mRNA expression levels of the downstream microphthalmia-associated transcription factor (MITF) and tyrosinase in B16F10 cells. Our data elucidate that bergaptol may promote melanogenesis by regulating the p-P38 and p-ERK signaling pathway.
CONCLUSIONS: This study will lay a foundation for discovering potential new drugs for treating vitiligo and provide feasible ideas for exploring the mechanism of traditional Chinese medicine.
OBJECTIVE: To explore the active substances and action mechanisms of PF in promoting melanogenesis.
METHODS: Firstly, UPLC-UV-Q-TOF/MS was used to characterize the components in PF extract and identify the absorption components and metabolites of PF after oral administration at usual doses in rats. Secondly, the active substances and related targets and pathways were predicted by network pharmacology and molecular docking. Finally, pharmacodynamic and molecular biology experiments were used to verify the prediction results.
RESULTS: The experimental results showed that 15 compounds were identified in PF extract, and 44 compounds, consisting of 8 prototype components and 36 metabolites (including isomers) were identified in rats\' plasma. Promising action targets (MAPK1, MAPK8, MAPK14) and signaling pathways (MAPK signaling pathway) were screened and refined to elucidate the mechanism of PF against vitiligo based on network pharmacology. Bergaptol and xanthotol (the main metabolites of PF), psoralen (prototype drug), and PF extract significantly increased melanin production in zebrafish embryos. Furthermore, bergaptol could promote the pigmentation of zebrafish embryos more than psoralen and PF extract. Bergaptol significantly increased the protein expression levels of p-P38 and decreased ERK phosphorylation in B16F10 cells, which was also supported by the corresponding inhibitor/activator combination study. Moreover, bergaptol increased the mRNA expression levels of the downstream microphthalmia-associated transcription factor (MITF) and tyrosinase in B16F10 cells. Our data elucidate that bergaptol may promote melanogenesis by regulating the p-P38 and p-ERK signaling pathway.
CONCLUSIONS: This study will lay a foundation for discovering potential new drugs for treating vitiligo and provide feasible ideas for exploring the mechanism of traditional Chinese medicine.
摘要：
背景：根据《本草纲目》(李世珍，明朝)和福利药房(宋朝)，补骨脂（PF），传统中药（TCM）具有苦味和温暖的性质，具有治疗脾肾虚和皮肤病的功效。尽管PF自古以来就被广泛使用，并在治疗白癜风方面显示出令人满意的疗效，PF促进黑素生成的活性物质和机制尚不清楚。
目的：探讨PF促进黑素生成的活性物质及其作用机制。
方法：首先，UPLC-UV-Q-TOF/MS用于表征PF提取物中的成分，并鉴定大鼠口服常规剂量后PF的吸收成分和代谢产物。其次,通过网络药理学和分子对接对活性物质及相关靶点和通路进行预测。最后,药效学和分子生物学实验验证了预测结果。
结果：实验结果表明，在PF提取物中鉴定出15种化合物，和44种化合物，在大鼠血浆中鉴定出8种原型成分和36种代谢物(包括异构体)。筛选并提炼有希望的作用靶点（MAPK1、MAPK8、MAPK14）和信号通路（MAPK信号通路），以网络药理学为基础阐明PF抗白癜风的作用机制。Bergaptol和xanthotol（PF的主要代谢产物），补骨脂素（原药），和PF提取物显着增加斑马鱼胚胎中黑色素的产生。此外，与补骨脂素和PF提取物相比，贝尔加普醇对斑马鱼胚胎色素沉着的促进作用更强。Bergaptol能显著提高B16F10细胞中p-P38蛋白的表达水平,降低ERK的磷酸化,这也得到了相应的抑制剂/激活剂组合研究的支持。此外，bergaptol增加了B16F10细胞中下游小眼症相关转录因子（MITF）和酪氨酸酶的mRNA表达水平。我们的数据阐明了bergaptol可能通过调节p-P38和p-ERK信号通路来促进黑素生成。
结论：本研究将为发现治疗白癜风的潜在新药奠定基础，为探索中医药作用机制提供可行思路。
目的：探讨PF促进黑素生成的活性物质及其作用机制。
方法：首先，UPLC-UV-Q-TOF/MS用于表征PF提取物中的成分，并鉴定大鼠口服常规剂量后PF的吸收成分和代谢产物。其次,通过网络药理学和分子对接对活性物质及相关靶点和通路进行预测。最后,药效学和分子生物学实验验证了预测结果。
结果：实验结果表明，在PF提取物中鉴定出15种化合物，和44种化合物，在大鼠血浆中鉴定出8种原型成分和36种代谢物(包括异构体)。筛选并提炼有希望的作用靶点（MAPK1、MAPK8、MAPK14）和信号通路（MAPK信号通路），以网络药理学为基础阐明PF抗白癜风的作用机制。Bergaptol和xanthotol（PF的主要代谢产物），补骨脂素（原药），和PF提取物显着增加斑马鱼胚胎中黑色素的产生。此外，与补骨脂素和PF提取物相比，贝尔加普醇对斑马鱼胚胎色素沉着的促进作用更强。Bergaptol能显著提高B16F10细胞中p-P38蛋白的表达水平,降低ERK的磷酸化,这也得到了相应的抑制剂/激活剂组合研究的支持。此外，bergaptol增加了B16F10细胞中下游小眼症相关转录因子（MITF）和酪氨酸酶的mRNA表达水平。我们的数据阐明了bergaptol可能通过调节p-P38和p-ERK信号通路来促进黑素生成。
结论：本研究将为发现治疗白癜风的潜在新药奠定基础，为探索中医药作用机制提供可行思路。
