大黄酚及P38阻断剂对紫外线诱导人永生化表皮细胞光老化的影响及机制研究

目的 研究大黄酚及P38阻断剂SB203580抑制中波紫外线诱导的人永生化表皮细胞(HaCaT immortalizedhuman epidermal cells,HaCaT)光老化的作用及其作用机制.方法 采用MTT法检测不同浓度的大黄酚及SB203580对HaCaT细胞增殖的影响;将细胞按随机数字表法分为空白组、模型组、大黄酚组、P38阻断剂组.培养24 h后,大黄酚组及P38阻断剂组分别加入10-6 mol/L大黄酚及10-7 mol/L SB203580溶液.继续培养24 h,除空白组外,其他各组细胞于紫外灯下照射,照射强度0.61 mW/cm2,照射时间7 min,照射距离10 cm.继续培养24 h后,采用MTT法检测各组细胞的增殖活性;Western Blot检测各组细胞中P38、磷酸化P38(P-P38)、TNF-α 及IL-6蛋白表达.结果 与空白组比较,模型组HaCaT细胞增殖率降低(P<0.01);与模型组比较,大黄酚组及P38阻断剂组HaCaT细胞增殖率升高(P<0.01);与模型组比较,大黄酚组及P38阻断剂组细胞P-P38(0.419±0.029)、(0.398±0.015)比(0.497±0.051)]、TNF-α(0.435±0.025)、(0.411±0.021)比(0.509±0.040)]及IL-6(0.457±0.027)、(0.432±0.018)比(0.478±0.036)]蛋白表达降低(P<0.01).结论 大黄酚及SB203580可抑制紫外线诱导的HaCaT细胞光老化,其作用机制可能与抑制P38MAPK通路及炎症因子表达有关.

The mechanism research of Rhubarb phenol and P38 inhibitor suppression on light aging of human skin HaCaT induced by UVB

Objective To study the rhubarb phenol and P38 inhibitor suppression on light aging of human skin HaCaT induced by UVB. Methods The potential of cell proliferation of the different concentrations of rhubarb phenol and P38 inhibitor (SB203580) on human skin HaCaT was detected by MTT method. The cells was divided into the control group, the model group, the rhubarb phenol group, the SB203580 group by random number table method after the 24 h incubation. The 10-6 mol/L rhubarb phenol and 10-7 mol/L SB203580 were added to the rhubarb phenol group and SB203580 group for 24h, The competence of cultured cell proliferation which was irradiated with UVB of intensity of 0.61mW/cm2, mutiply time of 7 min and distance of 10 cm for 24 h except the control group; Western Blot method detected rhubarb phenol and P38 inhibitor of the influence of P38, P-P38, TNF-α, IL-6 protein. Results Compared with control group, the cell proliferation in UVB group significantly reduced (P<0.01); Compared with UVB group, the expression of the P-P38 (0.419 ± 0.029, 0.398 ± 0.015 vs. 0.497 ± 0.051), TNF-α (0.435 ± 0.025, 0.411 ± 0.021 vs. 0.509 ± 0.040) and IL-6 (0.457 ± 0.027, 0.432 ± 0.018 vs. 0.478 ± 0.036) in rhubarb phenol and P38 inhibitor group significantly reduced (P<0.01). Conclusions The rhubarb phenol and P38 inhibitor could significantly suppress HaCaT cells light aging, and its mechanism may be related with inhibiting P38 signaling pathways, and inhibiting the secretion of inflammatory cytokines.