二苯乙烯苷可抑制中波紫外线诱导的人皮肤成纤维细胞光老化

Stilbene glucoside inhibits ultraviolet radiation B-induced photoaging in human skin fibroblasts

目的:研究二苯乙烯苷(TSG)对紫外线B(UVB)诱导的人皮肤成纤维细胞(HSF)应激性提早衰老的作用及可能的机制。方法:UVB小剂量、多次照射HSF,每次照射完毕后分别用0.02、0.10、0.50 mmol/L浓度的TSG处理。实验分为6组:空白对照组、模型对照组、UVB+0.02 mmol/L TSG组、UVB+0.10 mmol/L TSG组、UVB+0.50 mmol/L TSG组、TSG对照组。采用细胞计数法(CCK-8)检测细胞增殖活性;细胞化学染色法检测衰老相关β-半乳糖苷酶(SA-β-gal)表达;TBA法、WST-1法测定细胞丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性;ELISA法测定细胞上清液中基质金属蛋白酶-1(MMP-1)的含量变化。结果:与空白对照组比较,模型对照组细胞增殖活性减弱(P<0.05),SA-β-gal染色呈强阳性,细胞裂解液中SOD活性减弱、MDA含量增加,MMP-1浓度升高(P<0.05或P<0.01)。与模型对照组比较,UVB+各浓度TSG组细胞增殖活性改善(均P<0.05),SA-β-gal染色阳性率下降,细胞裂解液中SOD活性增强、MDA含量减少,MMP-1浓度减小(P<0.05或P<0.01)。结论:TSG可以在一定程度上抑制UVB诱导的HSF应激性提早衰老,该作用可能与改善氧化应激、抑制MMP-1高表达有关。

Objective:To study the effects of tetrahydroxy stilbene-2-O-β-D-glucoside(TSG)on stress-induced premature senescence of human skin fibroblasts(HSF)exposed to ultraviolet radiation B(UVB)and its possible mechanism.Methods:HSFs were repeatedly exposed to UVB at a subcytotoxic level.TSG treatment(0.02,0.10 and 0.50 mmol/L)was given immediately after each irradiation.The HSFs were divided into six groups:blank control group,model group,UVB+0.02 mmol/L TSG group,UVB+0.10 mmol/L TSG group,UVB+0.50 mmol/L TSG and TSG group(0.50 mmol/L).Cell counting kit-8(CCK-8)was used to evaluate the proliferative activity of cells;senescence-associated-β-galactosidase(SA-β-gal)staining was performed to estimate the degree of premature senescence in cells;TBA method and WST-1 method were used to detect intracellular malondialdehyde(MDA)and superoxide dismutase(SOD)activities;and ELISA was applied to quantify the secretion level of matrix metalloproteinase1(MMP-1)in cultured supernatant.Results:Compared with the blank control group,the proliferative activity and SOD level in the model group decreased(P<0.05),while the percentage of SA-β-gal-positive cells,MDA and MMP-1 levels increased(P<0.05 or P<0.01).Compared with the model group,the proliferative activity and SOD level increased in UVB+TSG groups(all P<0.05),and the percentage of SA-β-gal-positive cells,MDA and MMP-1 levels decreased(P<0.05 or P<0.01).Conclusion:TSG can inhibit UVB-induced premature senescence of HSF,which may be related to the inhibition of oxidative stress and high expression of MMP-1.