飞燕草素对光化学损伤661W细胞的保护作用及其机制

目的 探讨飞燕草素对光化学损伤661W细胞的保护作用及其机制.方法 取661W细胞进行培养,根据预实验结果采用(2000±200)lux光照强度持续照射细胞48 h作为造模条件,采用5 μmol·L-1飞燕草素、3 mmol·L-1抗氧化剂N-乙酰-L-半胱氨酸(NAC)为最佳用药浓度.细胞分组如下:对照组,常规避光培...

Protective effect of delphinidin on photochemical damaged 661W cells and its mechanisms

Objective　To investigate the protective effect and mechanism of delphinidin on photochemical damaged 661W cells.Methods　The 661W cells were selected for culture. According to the results of the preliminary experiment, the cells were continuously irradiated with (2000±200)lux light intensity for 48 hours as the model condition, and 5 μmol·L^-1 delphinidin and 3 mmol·L^-1 antioxidant N-acetyl-L-cysteine (NAC) were used as the preferred drug concentration. The cells were grouped as follows:control group, in which cells were cultured routinely for 48 hours in the dark; light group, in which cells were cultured in (2000±200) lux light for 48 hours; light delphinidin group, in which cells were cultured in (2000±200) lux light for 24 hours, then to the culture medium containing 5 μmol·L^-1 delphinidin for 24 hours; dark delphinidin group, in which cells were cultured in the dark for 24 hours, then to the culture medium containing 5 μmol·L^-1 delphinidin for 24 hours; light NAC group, in which cells were cultured in (2000±200) lux light for 24 hours, then to the culture medium containing 3 mmol·L^-1 NAC for 24 hours.Results　Microscopically, the cells in control group grew well, and the cells in the light group wrinkled and curled and the number of exfoliated cells increased. CCK-8 results showed that compared with the control group, the survival rates of cells in the light group was significantly reduced, and the difference was statistically significant (P<0.05). There was no significant change in cell survival rate of the dark delphinidin group (P>0.05). Compared with the light group, the cell survival rate of the light delphinidin group, the light NAC group and the dark delphinidin group increased significantly, and the differences were all statistically significant (all P<0.05). Compared with the control group, the ROS content of the light group was significantly increased, and the difference was statistically significant (P<0.05). The ROS content in the dark delphinidin group was not significantly reduced, and the difference was not statistically significant (P>0.05). Compared with the light group, the ROS content of the light delphinidin group, the light NAC group and the dark delphinidin group were all decreased significantly, and the differences were statistically significant (all P<0.05). Compared with the control group, the mitochondrial membrane potential for the light group decreased significantly, and the difference was statistically significant (P<0.05). There was no significant change in mitochondrial membrane potential in the dark delphinidin group, and the difference was not statistically significant (P>0.05). Compared with the light group, the mitochondrial membrane potential for the light delphinidin group, the light NAC group and the dark delphinidin group all increased significantly, and the differences were statistically significant (all P<0.05). Compared with the light group, the apoptosis rate in the light delphinidin group, the light NAC group and the dark delphinidin group all decreased significantly, and the differences were statistically significant (all P<0.05). Compared with the light group, iNOS, Bax, Cytochrome C, and Cleaved-Caspase-3 protein expression in the light delphinidin group, the light NAC group and the dark delphinidin group were down-regulated, Bcl-2 protein expression were up-regulated, and Bcl-2/Bax values were significantly increased, and all these differences were statistically significant (all P<0.05).Conclusion　Delphinidin can protect 661W cells from photochemical damage by regulating the expression of important factors related to oxidative stress mitochondrial apoptosis pathways.