目的:评价二氢杨梅素(DMY)对小鼠黑色素瘤细胞 B16 的影响,并揭示其抑制黑色素产生的作用机制.方法:CCK8 法筛选DMY对B16 细胞增殖活性影响的最佳浓度.分为对照组、模型组(α-MSH)阳性药组(α-MSH+熊果苷)、给药组(α-MSH+DMY),采用 CCK8 法测定不同组别 B16 细胞的增殖活性,ELISA法检测B16 细胞中黑色素和酪氨酸酶的含量,透射电镜观察细胞中黑素小体数量,分子对接法预测DMY 降低黑色素含量作用的蛋白,免疫荧光法和 Western blot 分别测α-MSH和MITF蛋白的表达.结果:DMY抑制B16 细胞增殖的最佳浓度为 62.5μmol·L-1.最佳浓度下,与模型组相比,DMY可以抑制B16 细胞的增殖,降低黑色素和酪氨酸酶含量,减少黑素小体数目,抑制α-MSH 和MITF蛋白表达.结论:二氢杨梅素通过调节α-MSH/MITF通路抑制B16 细胞产生黑色素.
Dihydromyricetin through α-MSH/MITF pathway inhibits melanin production
Objective:To evaluate the effect of dihydromyricetin(DMY)on mouse melanoma cell B16 and reveal its mechanism of inhibiting melanin production.Method:The CCK8 method was used to screen the optimal concentration of DMY on the proliferation activity of B16 cells.Divided into control group and model group(α-MSH)positive drug group(α-MSH+arbutin),administration group(α-MSH+DMY),the proliferation activity of B16 cells in different groups was measured using the CCK8 method,the content of melanin and tyrosinase in B16 cells was detected using ELISA method,the number of melanosomes in cells was observed using transmission electron microscopy,and the molecular docking method was used to predict the protein that DMY can reduce melanin content.Immunofluorescence and Western blot were used to measure the levels of melanin and tyrosinase in B16 cells,the expression of α-MSH and MITF proteins.Result:The optimal concentration for DMY to inhibit B16 cell proliferation is 62.5 μmol·L-1.At the optimal concentration,compared with the model group,DMY can inhibit the proliferation of B16 cells,reduce melanin and tyrosinase content,reduce the number of melanosomes,and inhibitα-MSH and MITF protein expression.Conclusion:Dihydromyricetin regulates α-The MSH/MITF pathway inhibits the production of melanin in B16 cells.
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