摘要
目的探究枸杞多糖(LBP)对UVA诱导细胞黑色素生成的影响。方法用不同浓度LBP(0、20、50、100、200、300、400、500μg/mL)处理HaCaT细胞和HEM细胞,采用CCK8法检测对细胞增殖的影响,筛选出适宜浓度的LBP进行后续实验。使用300μg/mL LBP、20μM N-1A处理被30 J/cm~2 UVA照射的HaCaT细胞和HEM细胞,多巴氧化法测定细胞中tyrosinase活性、NaOH裂解法测定黑色素含量。通过酶联免疫吸附试验(ELISA)检测LBP对细胞内α-MSH蛋白表达的调控,应用Western blot检测细胞黑色素生成相关蛋白MITF、tyrosinase、TRP1和TRP2的表达水平。结果CCK8实验结果提示,当LBP浓度等于300μg/mL时,HaCaT细胞和HEM细胞的增殖无明显变化(t=1.89、0.07,P=0.107、0.947)。相较于UVA组,LBP在体外可以明显抑制UVA诱导的HEM细胞中tyrosinase活性(t=33.19,P=0.001)以及减低黑色素含量(t=7.13,P=0.019);ELISA结果提示,UVA照射可以促进HaCaT细胞和HEM细胞的α-MSH蛋白分泌(t=-3.79、-3.41,P=0.043、0.046),LBP能够抑制HaCaT细胞和HEM细胞中α-MSH的蛋白表达水平(t=3.75、3.41,P=0.044、0.048);Western blot结果显示,LBP可显著下调UVA诱导的黑色素生成相关蛋白MITF、tyrosinase、TRP1和TRP2的表达。结论LBP可显著抑制UVA诱导的细胞黑色素生成,其机制可能通过调控HaCaT细胞和HEM细胞中α-MSH蛋白水平实现。
Objective To investigate the effect of lycium barbarum polysaccharide(LBP)on UVA-induced melanogenesis.Methods HaCaT cells and HEM cells were treated with different concentrations of LBP(0,20,50,100,200,300,400,500μg/mL).CCK8 assay was performed to assess cell proliferation,in order to obtain the most suitable concentration of LBP for subsequent study.HaCaT cells and HEM cells were treated with 300μg/mL of LBP and(or)20μM N-1 A after irradiation with 30 J/cm~2 UVA.Tyrosinase activity was determined by L-dopa oxidation method and melanin content by NaOH analysis method.The intracellularα-MSH protein expression was measured by enzyme-linked immunosorbent assay(ELISA),and the expression levels of cellular melanogenesis-related proteins,including MITF,tyrosinase,TRP1 and TRP2,were determined by Western blot.Results Based on the results of CCK8 assay,the most suitable concentration of LBP was 300μg/mL.Compared with UVA group,LBP significantly inhibited UVA-induced cellular tyrosinase activity(t=33.19,P=0.001)and melanin production(t=7.13,P=0.019).The ELISA results suggested that UVA irradiation promotedα-MSH protein secretion in HaCaT cells and HEM cells(t=-3.79、-3.41,P=0.043、0.046),and LBP inhibited expression levels ofα-MSH protein in HaCaT cells and HEM cells(t=3.75、3.41,P=0.044、0.048).The Western blot results showed that LBP significantly downregulated the expression of the melanogenesis-related proteins,including MITF,tyrosinase,TRP1,and TRP2 induced by UVA.Conclusion LBP significantly inhibits UVA-induced melanogenesis,via regulating expression levels ofα-MSH protein in HaCaT cells and HEM cells.
作者
肖常青
陈教全
唐奕
欧珊珊
李华平
彭丽倩
李振洁
朱慧兰
XIAO Changqing;CHEN Jiaoquan;TANG Yi;OU Shanshan;LI Huaping;PENG Liqian;LI Zhenjie;ZHU Huilan(Guangzhou Institute of Dermatology,Guangzhou 510095,China)
出处
《皮肤性病诊疗学杂志》
2022年第5期406-411,共6页
Journal of Diagnosis and Therapy on Dermato-venereology
基金
广东省医学科学技术研究基金(B2020058)
广州市卫生健康项目(20201A010049)。
关键词
枸杞多糖
紫外线
Α-MSH
黑色素
lycium barbarum polysaccharide
ultraviolet light A
α-MSH
melanin
