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绿茶中多酚类物质60对斑马鱼和A375人黑色素瘤细胞中黑色素及酪氨酸酶的抑制作用

The Inhibitory Effect of Polyphenon 60 from Green Tea on Melanin and Tyrosinase in Zebrafish and A375 Human Melanoma Cells.

作者信息

Kazi Mehar Ali, Sahito Reshma, Abbas Qamar, Ullah Sana, Majid Abdul, Phull Abdul Rehman, Rahman Md Mominur, Kim Song Ja

机构信息

Institute of Biochemistry, University of Sindh, Jamshoro 76080, Pakistan.

Department of Zoology, University of Sindh, Jamshoro 76080, Pakistan.

出版信息

Evid Based Complement Alternat Med. 2022 Sep 2;2022:7739023. doi: 10.1155/2022/7739023. eCollection 2022.

Abstract

Polyphenon 60 (PP60) from green tea has long been used as an antioxidant, anticancer, antimicrobial, and antimutagenic. . To investigate tyrosinase inhibition-related kinetic mechanism and antimelanogenesis potential of PP60. . The effect of PP60 on melanin and tyrosinase was evaluated in A375 melanoma cells and zebrafish embryos. The melanoma cells were treated with 20, 40, and 60 g/mL of PP60, and tyrosinase expression was induced by using L-DOPA. The western blot method was used for the evaluation of tyrosinase expression. Cell lysates were prepared from treated and untreated cells for cellular tyrosinase and melanin quantification. Furthermore, zebrafish embryos were treated with 20, 40, and 60 g/mL of PP60 and reference drug kojic acid for determination of depigmentation and melanin quantification. assays were also performed to examine the impact of PP60 on mushroom tyrosinase activity. To determine cytotoxicity, MTT was used against melanoma cell line A375. . PP60 showed good tyrosinase inhibitory activity with an IC value of 0.697 ± 0.021 g/mL as compared to kojic acid a reference drug with an IC value of 2.486 ± 0.085 g/mL. Kinetic analysis revealed its mixed type of inhibition against mushroom tyrosinase. In addition, western blot analysis showed that at 60 g/mL dose of PP60 significantly reduced L-DOPA-induced tyrosinase expression in melanoma cells. PP60 significantly inhibits the cellular tyrosinase ( < 0.05) and reduces the melanin ( < 0.05) contents of melanoma cells. Furthermore, PP60 was found to be very potent in significantly reducing the zebrafish embryos' pigmentation ( < 0.05) and melanin ( < 0.05) content at the dose of 60 g/mL. . Our results demonstrate that PP60 has a strong potency to reduce pigmentation. It may be useful for the cosmetic industries to develop skin whitening agents with minimal toxic effects.

摘要

绿茶中的茶多酚60(PP60)长期以来一直被用作抗氧化剂、抗癌剂、抗菌剂和抗诱变剂。为了研究PP60与酪氨酸酶抑制相关的动力学机制及其抗黑色素生成潜力。在A375黑色素瘤细胞和斑马鱼胚胎中评估了PP60对黑色素和酪氨酸酶的影响。用20、40和60μg/mL的PP60处理黑色素瘤细胞,并使用L-多巴诱导酪氨酸酶表达。采用蛋白质免疫印迹法评估酪氨酸酶表达。从处理过和未处理的细胞中制备细胞裂解物,用于细胞酪氨酸酶和黑色素定量。此外,用20、40和60μg/mL的PP60以及参考药物曲酸处理斑马鱼胚胎,以测定色素沉着和黑色素定量。还进行了实验以研究PP60对蘑菇酪氨酸酶活性的影响。为了确定细胞毒性,将MTT用于黑色素瘤细胞系A375。与曲酸(IC值为2.486±0.085μg/mL的参考药物)相比,PP60表现出良好的酪氨酸酶抑制活性,IC值为0.697±0.021μg/mL。动力学分析表明其对蘑菇酪氨酸酶的抑制作用为混合型。此外,蛋白质免疫印迹分析表明,在60μg/mL剂量下,PP60显著降低了L-多巴诱导的黑色素瘤细胞中酪氨酸酶的表达。PP60显著抑制细胞酪氨酸酶(P<0.05),并降低黑色素瘤细胞的黑色素含量(P<0.05)。此外,发现PP60在60μg/mL剂量下能非常有效地显著降低斑马鱼胚胎的色素沉着(P<0.05)和黑色素含量(P<0.05)。我们的结果表明,PP60具有很强的减少色素沉着的能力。这对于化妆品行业开发毒性最小的皮肤美白剂可能是有用的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d5f4/9463005/379931791533/ECAM2022-7739023.001.jpg

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