Toxicological and melanin synthesis effects of Polygonum multiflorum root extracts on zebrafish embryos and human melanocytes
Toxicological and melanin synthesis effects of Polygonum multiflorum root extracts on zebrafish embryos and human melanocytes
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DOI：
10.7603/s40730-016-0042-4
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发表时间：
2016-01-01
影响因子：
0.9
通讯作者：
Nguyen Dinh Thang
中科院分区：
文献类型：
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作者：
Dang Thi Hoai Thanh;Nguyen Lai Thanh;Nguyen Dinh Thang
简介：何首乌（Polygonum multiflorum，PM）通常被用作治疗各种疾病的民间药物，例如人类头发的早期灰白。然而，在体外和体内评价PM的毒理学和生物学效应的研究有限。在这项研究中，PM根提取物在乙酸乙酯（PM-E）和蒸馏水（PM-W）的影响，其对致畸缺陷/死亡的发展进行了检查。此外，评价了它们对人黑素细胞中黑色素形成和野生型品系AB斑马鱼（Danio rerio）胚胎/幼体中色素沉着的影响。方法：采用极性递增的溶剂提取紫云英根中的植物化学物质。采用化学暴露试验研究了紫云英根提取物对斑马鱼胚胎/仔鱼的毒性效应。通过分光光度计使用415和400 nm的双波长测量斑马鱼胚胎/幼虫以及人黑色素细胞中形成的总黑色素。通过实时PCR分析基因表达。所有实验独立重复三次并进行统计分析。结果如下：结果表明，40 mg/L和105 mg/L的黄芪提取物可诱导受精后4 d的斑马鱼胚胎出现卵黄囊水肿（或心脏水肿）、血管畸形、坏死和躯干畸形等畸形缺陷，乙酸乙酯提取物和蒸馏水提取物的致畸指数（TI）分别为1.43和0.63。我们的研究结果还表明，PM-W显着增加的色素沉着水平的胚胎/幼虫和诱导黑色素形成的人黑素细胞。PM-W暴露的胚胎/幼虫中黑色素的量分别是对照胚胎/幼虫和对照黑色素细胞中黑色素的量的2.2倍和1.71倍。结论：我们的研究还表明，PM-W胚胎/幼虫中色素沉着水平的增加或黑素细胞中黑素生物合成的增加都受到酪氨酸酶活化的调节。总之，我们的研究表明，PM根提取物可用作治疗早期头发变白以及与色素沉着丧失相关的各种其他疾病的潜在药物。然而，这些PM根提取物也可能对胚胎产生一些负面影响;因此，在怀孕期间使用时应小心。
Introduction: Polygonum multiflorum (PM) has been commonly used as folk medicine for treatment of various conditions, such as early graying of hair in humans. However, there have been limited studies which have evaluated the toxicological and biological effects of PM in vitro as well as in vivo. In this study, PM root extracts in ethyl acetate (PM-E) and in distilled water (PM-W) were examined for their effects on the development of teratogenic defects/deaths. Additionally, they were evaluated for their effects on melanin formation in human melanocytes and pigmentation in embryos/larvae of wild type strain AB zebrafish (Danio rerio). Methods: Phytochemicals in PM roots were extracted in several solvents with increasing in their polarities. Chemical exposure tests were performed to investigate the toxic effects of PM root extracts on Zerbrafish embryos/larvae. Total melanins formed in zebrafish embryos/larvae as well as in human melanocytes were measured by spectrophotometer using dual wavelengths of 415 and 400 nm. Gene expressions were analyzed by realtime PCR. All experiments were independently repeated three times and statistical analyzed. Results: Our results showed that PM root extracts at concentrations of 40 mg/L and 105 mg/L induced the development of teratogenic defects, including yolk sac edema (or heart edema), hemovascular defects, necrosis and abnormal trunk in zebrafish embryos at 4 days post fertilization; teratogenic indexes (TIs) were 1.43 and 0.63 for ethyl acetate extract and distilled water extract, respectively. Our results also demonstrated that PM-W significantly increased the pigmentation level of embryos/larvae and induced melanin formation in human melanocytes. The amount of melanin in PM-W-exposed embryos/larvae was 2.2-fold and 1.71-fold greater than those in the control embryos/larvae and control melanocytes, respectively. Conclusion: Our study also showed that the increased level of pigmentation in PM-W embryos/larvae or melanin biosynthesis in melanocytes were both regulated by activation of tyrosinase. Conclusively, our study suggests that PM root extracts could be used as potential agents for treatment of early hair graying as well as various other diseases related to loss of pigmentation. However, these PM root extracts may also have some negative effects on embryos; therefore it should be careful when using for women during pregnancy.