台湾白及愈伤组织提取物通过Nrf2/HO-1信号通路提高细胞抗氧化活性并抑制斑马鱼的黑色素生成。
Extract of Bletilla formosana callus elevates cellular antioxidative activity via Nrf2/HO-1 signaling pathway and inhibits melanogenesis in zebrafish.
作者信息
Wang Ruei-Ting, Yen Jui-Hung, Liao Yi-Chi, Li Yi-Zhen, Wang Wei-Ping
机构信息
CHLITINA Research and Development Center, CHLITINA Holding Ltd., Taipei, 110050, Taiwan.
Biomedical Technology and Device Research Laboratories, Industrial Technology Research Institute, Hsinchu, 310401, Taiwan.
出版信息
J Genet Eng Biotechnol. 2023 Mar 6;21(1):26. doi: 10.1186/s43141-023-00482-0.
BACKGROUND
Bletilla species are endangered terrestrial orchids used in natural skin care formulas in Asia for a long history. In order to explore the bioactivity potential of Bletilla species as a cosmetic ingredient in a sustainable resource manner, the callus of Bletilla formosana (Hayata) Schltr. was established and extracted by an eco-friendly supercritical fluid CO extraction (SFE-CO) method. The intracellular reactive oxygen species (ROS) scavenging activity and antioxidation-related gene expression of the callus extract were evaluated in both Hs68 fibroblast cells and HaCaT keratinocytes. The melanogenesis-inhibitory effect was investigated in B16F10 melanoma cells and in an in vivo zebrafish model.
RESULTS
The calli of B. formosana were propagated for 10-15 generations with a consistent yellow friable appearance and then subjected to SFE-CO extraction to obtain a yellow pasty extract. Obvious intracellular ROS scavenging activity of the extract was detected in both Hs68 and HaCaT cells with 64.30 ± 8.27% and 32.50 ± 4.05% reduction at the concentration of 250 μg/mL. Moreover, marked expression levels of heme oxygenase-1 (HO-1) and (NAD(P)H) quinone oxidoreductase-1 (NQO1) genes were detected after 6-h and 24-h treatments. These results indicate the cellular antioxidative activity of B. formosana callus extract was probably activated via the nuclear factor erythroid 2-related factor 2 (Nrf2)/HO-1 signaling pathway. Melanogenesis-inhibitory effect of the extract was observed in α-MSH stimuli-inducing B16F10 cells with 28.46% inhibition of intracellular melanin content at the concentration of 50 μg/ml. The effect was confirmed with in vivo zebrafish embryos that showed a relative pigmentation density of 80.27 ± 7.98% at the concentration of 100 μg/mL without toxicity.
CONCLUSION
Our results shed light on a sustainable utilization of Bletilla species as a potential ingredient for skin.
背景
白及属植物是濒危的陆生兰花，在亚洲被用于天然护肤品配方已有很长历史。为了以可持续资源的方式探索白及属植物作为化妆品成分的生物活性潜力，建立了台湾白及（Bletilla formosana (Hayata) Schltr.）的愈伤组织，并采用环保的超临界流体CO₂萃取（SFE-CO₂）方法进行提取。在Hs68成纤维细胞和HaCaT角质形成细胞中评估了愈伤组织提取物的细胞内活性氧（ROS）清除活性和抗氧化相关基因表达。在B16F10黑色素瘤细胞和体内斑马鱼模型中研究了其抑制黑色素生成的作用。
结果
台湾白及愈伤组织传代培养10 - 15代，外观始终呈黄色易碎状，然后进行SFE-CO₂萃取以获得黄色糊状提取物。在Hs68和HaCaT细胞中均检测到提取物具有明显的细胞内ROS清除活性，在浓度为250μg/mL时，清除率分别为64.30±8.27%和32.50±4.05%。此外，在处理6小时和24小时后，检测到血红素加氧酶-1（HO-1）和（NAD(P)H）醌氧化还原酶-1（NQO1）基因的显著表达水平。这些结果表明，台湾白及愈伤组织提取物的细胞抗氧化活性可能是通过核因子红细胞2相关因子2（Nrf2）/HO-1信号通路激活的。在α-MSH刺激诱导的B16F10细胞中观察到提取物的黑色素生成抑制作用，在浓度为50μg/ml时，细胞内黑色素含量抑制率为28.46%。在体内斑马鱼胚胎中证实了该作用，在浓度为100μg/mL时，相对色素沉着密度为80.27±7.98%，且无毒性。
结论
我们的研究结果为白及属植物作为皮肤潜在成分的可持续利用提供了依据。
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