橙皮苷对UVB照射后的B16和HaCaT细胞的作用

Effect of hesperidin on B16 and HaCaT cell lines irradiated by narrowband-UVB light

目的:研究橙皮苷对紫外线照射后的鼠黑素瘤B16和永生化人角质形成细胞HaCaT株细胞功能的影响。方法:采用四甲基偶氮唑蓝(MTT)法测定细胞的增殖,NaOH法测定黑素含量,Takahashi法测定酪氨酸酶活性;流式细胞仪分析HaCaT细胞的CXC型趋化因子受体(CXCR)2表达;ELISA法测定细胞培养液中转化生长因子(TGF)-β1含量的变化。结果:50～600mg/L橙皮苷对B16和HaCaT细胞的增殖活性无明显影响,200mg/L浓度以上时轻度抑制酪氨酸酶活性和黑素合成。不同浓度的橙皮苷作用后,角质形成细胞培养上清中TGF-β1水平呈无规律性变化。正常角质形成细胞的CXCR2表达不受橙皮苷的影响,紫外线照射后,CXCR2表达增加,200～600mg/L橙皮苷以浓度依赖方式抑制其表达,浓度在600mg/L时作用更显著。结论:橙皮苷无细胞毒性,其作用温和,具多效性,是有前途的褪色和光防护制剂。

Objective： To investigate the effects of hesperidine on B16 and HaCat cell lines irradiated by Narrowband-UVB light. Methods： MTT, NaOH assay, Takahashi technique were applied to measure the proliferation of B16 and HaCaT cells, the synthesis of melanin, and the tyrosinase activity of B16 cell respectively. The expression of CXC receptor 2 （CXCR2） on keratinocytes was analyzed by flow cytometry. To detect the contents of transforming growth factor-β1 （TGF-β1）, enzyme linked immunosorbent assay was applied. Results： The findings indicated that hesperidine at the concentrations of 50-600 mg/L did not affect the proliferation of B16 and HaCaT cells. At the doses of 200 mg/L and over, hesperidine inhibited the tyrosinase activity and melanogenesis in B16 cells. The levels of TGF-β1 in the supematant of cultured keratinocytes presented as ＂wave shape＂ according to hesperidine contents. The expression of CXCR2 in the normal cultured keratinocytes did not change with different concentrations of hesperidine. However, the expression of CXCR2 in the cultured cells irradiated by narrowband-UVB light was increased obviously. When 200-600 mg/L hesperidine were added to the media, in which the cultured keatinocytes would be irradiated by Narrowband-UVB light later, the expression of CXCR2 decreased markedly in a dose-dependent manner, especially, when the dosage of hesperidine was 600 mg/L. Conclusions： Hesperidine has various biological effects, with gentle effectiveness, and without cytotoxicity. It is a potent plant extract for whitening and photoprotection.