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研究葛根素(puerarin,PUE)对UVB诱导的HaCaT细胞DNA损伤的影响和作用机制。葛根素预处理细胞以30 mJ.cm2 UVB照射,24 h后以彗星电泳实验检测DNA损伤,薄层色谱法和气相色谱法检测神经酰胺,流式细胞术检测细胞内游离钙离子,Western blotting法检测p38蛋白磷酸化水平。结果显示,UVB模型组细胞DNA损伤、细胞内游离钙离子、神经酰胺、磷酸化p38蛋白显著增加;与UVB模型组相比,各葛根素预处理组的DNA损伤、细胞内游离钙离子、神经酰胺、磷酸化p38蛋白均减少。结果表明,葛根素首先抑制神经酰胺蓄积,阻止下游的细胞内钙离子升高和p38 MAPK通路激活,从而减轻UVB诱导的HaCaT细胞DNA损伤。
To investigate the effect and mechanism of puerarin (PUE) on DNA damage induced by UVB in HaCaT cells. The puerarin pretreated cells were exposed to UVB at 30 mJ · cm 2 for 24 h, DNA damage was detected by comet assay 24 h later, ceramide was detected by thin layer chromatography and gas chromatography, and intracellular free calcium ions were detected by flow cytometry. Western blotting Detection of p38 protein phosphorylation level. The results showed that DNA damage, intracellular free calcium, ceramide and phosphorylated p38 protein in UVB model group were significantly increased. Compared with UVB model group, DNA damage, intracellular free calcium, Amide, phosphorylated p38 protein are reduced. The results showed that puerarin inhibited the accumulation of ceramide firstly, prevented the intracellular calcium increase in the downstream and activated the p38 MAPK pathway, and reduced the UVB-induced DNA damage in HaCaT cells.