中国临床药理学杂志

目的 研究蟛蜞菊内酯(WEL)对人永生化角质形成细胞(HaCaT)氧化损伤保护作用。方法 将HaCaT细胞随机分为对照组(正常培养)、模型组(100μg·mL^-1氧化低密度脂蛋白ox-LDL)、中剂量实验组(100μg·mL^-1 ox-LDL+20μmol·L^-1 WEL)、高剂量实验组(100μg·mL^-1 ox-LDL+40μmol·L^-1 WEL)。用噻唑蓝(MTT)试剂盒检测细胞存活率，用细胞衰老β-半乳糖苷酶染色试剂盒检测细胞衰老情况，用Hoechst 33258染色法检测细胞凋亡情况，用蛋白质印迹实验检测细胞中衰老相关蛋白的表达情况。结果 对照组、模型组和中、高剂量实验组细胞存活率分别为(100.00±4.49)%、(163.99±11.37)%、(96.43±6.32)%和(93.02±9.50)%,细胞衰老占比率分别为(7.86±4.51)%、(25.42±5.13)%、(13.32±5.28)%和(9.71±2.37)%,细胞凋亡率分别为(5.36±1.22)%、(17.27±4.39)%、(12.89±2.49)%和(7.94±3.63)%,p21蛋白相对表达水平分别为0.79±0.33、1.68±0.65、1.09±0.45和0.75±0.31,p16蛋白相对表达水平分别为1.00±0.29、2.10±0.31、1.50±0.28和1.20±0.15。模型组的上述指标与对照组比较，中、高剂量实验组的上述指标与模型组比较，在统计学上差异均有统计学意义(P<0.000 1,P<0.001,P<0.05)。结论 WEL可以抑制HaCaT细胞氧化损伤，其机制与抑制损伤所诱导的衰老有关。

Objective To explore the protective effects of wedelolactone(WEL) against oxidative damage in HaCaT cells. Methods HaCaT cells were randomly divided into control group(normal culture),model group [100 μg·mL^-1 oxidized low-density lipoprotein(ox-LDL)],experimental-M group(100 μg·mL^-1 ox-LDL+20 μmol·L^-1 WEL) and experimental-H group(100 μg·mL^-1 ox-LDL+40 μmol·L^-1 WEL). Methyl thiazolyl tetrazolium(MTT) assay was used to detect cell survival rate; senescence β-galactosidase staining kit was used to detect cell senescence; Hoechst 33258 staining was used to detect the apoptosis of the cells; Western blot was used to detect the expression of senescence-related proteins in the cells. Results The cell survival rates of the control group, model group and experimental-M,-H groups were(100.00±4.49)%,(163.99±11.37)%,(96.43±6.32)% and(93.02±9.50)%,respectively; the proportions of cell senescence were(7.86±4.51)%,(25.42±5.13)%,(13.32±5.28)% and( 9. 71 ± 2. 37)%, respectively; the apoptosis rates were( 5. 36 ± 1. 22)%,( 17. 27 ± 4. 39)%,(12. 89 ± 2. 49)% and(7. 94 ± 3. 63)%, respectively; the relative expression levels of p21 were 0. 79 ± 0. 33,1. 68 ± 0. 65,1. 09 ± 0. 45 and 0. 75 ± 0. 31,respectively; the relative expression levels of p16 were 1. 00 ± 0. 29,2. 10 ± 0. 31,1. 50 ± 0. 28 and 1. 20 ± 0. 15,respectively. The above indexes of the model group were compared withthe control group,and the above indexes of the experimental-M,-H groups were compared with the model group(P<0. 000 1,P<0. 001,P<0. 05).Conclusion WEL can inhibit oxidative damage in HaCaT cells,and its mechanismis related to the inhibition of senescence induced by the damage.