Xiong Yin, Halima Mahmoud, Che Xiaoyan, Zhang Yiming, Schaaf Marcel J M, Li Minghui, Gao Min, Guo Liqun, Huang Yan, Cui Xiuming, Wang Mei
Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, China.
Institute of Biology Leiden, Leiden University, Leiden, Netherlands.
Front Pharmacol. 2022 Jul 7;13:946900. doi: 10.3389/fphar.2022.946900. eCollection 2022.
(PN) is a Chinese medicinal herb that is traditionally used to treat inflammation and immune-related diseases. Its major active constituents are saponins, the types and levels of which can be changed in the process of steaming. These differences in saponins are causally relevant to the differences in the therapeutic efficacies of raw and steamed PN. In this study, we have prepared the extracts of steamed PN (SPNE) with 70% ethanol and investigated their immunomodulatory effect using a zebrafish tail-fin amputation model. A fingerprint-effect relationship analysis was performed to uncover active constituents of SPNE samples related to the inhibitory effect on neutrophil number. The results showed that SPNE significantly inhibited the neutrophil number at the amputation site of zebrafish larvae. And SPNE extracts steamed at higher temperatures and for longer time periods showed a stronger inhibitory effect. Ginsenosides Rh, Rk, Rh, 20()-Rg, and 20()-Rg, of which the levels were increased along with the duration of steaming, were found to be the major active constituents contributing to the neutrophil-inhibiting effect of SPNE. By additionally investigating the number of neutrophils in the entire tail of zebrafish larvae and performing TUNEL assays, we found that the decreased number of neutrophils at the amputation site was due to both the inhibition of their migration and apoptosis-inducing effects of the ginsenosides in SPNE on neutrophils. Among them, Rh and 20()-Rg did not affect the number of neutrophils at the entire tail, suggesting that they only inhibit the migration of neutrophils. In contrast, ginsenosides Rk, Rh, 20()-Rg, and SPNE did not only inhibit the migration of neutrophils but also promoted neutrophilic cell death. In conclusion, this study sheds light on how SPNE, in particular the ginsenosides it contains, plays a role in immune modulation.
人参是一种传统上用于治疗炎症和免疫相关疾病的中草药。其主要活性成分是皂苷，在蒸煮过程中其类型和含量会发生变化。这些皂苷的差异与生人参和蒸制人参治疗效果的差异存在因果关系。在本研究中，我们用70%乙醇制备了蒸制人参提取物（SPNE），并使用斑马鱼尾鳍截肢模型研究了它们的免疫调节作用。进行了指纹-效应关系分析，以揭示与对中性粒细胞数量的抑制作用相关的SPNE样品的活性成分。结果表明，SPNE显著抑制斑马鱼幼虫截肢部位的中性粒细胞数量。在较高温度下蒸制较长时间的SPNE提取物显示出更强的抑制作用。人参皂苷Rh、Rk、Rh、20()-Rg和20()-Rg的含量随着蒸煮时间的延长而增加，它们被发现是导致SPNE对中性粒细胞抑制作用的主要活性成分。通过额外研究斑马鱼幼虫整个尾巴中的中性粒细胞数量并进行TUNEL分析，我们发现截肢部位中性粒细胞数量的减少是由于它们的迁移受到抑制以及SPNE中的人参皂苷对中性粒细胞的凋亡诱导作用。其中，Rh和20()-Rg不影响整个尾巴中的中性粒细胞数量，这表明它们仅抑制中性粒细胞的迁移。相比之下，人参皂苷Rk、Rh、20()-Rg和SPNE不仅抑制中性粒细胞的迁移，还促进中性粒细胞死亡。总之，本研究揭示了SPNE，特别是其所含的人参皂苷，在免疫调节中发挥作用的机制。
