飞燕草素对光化学损伤661W细胞的保护作用及其机制  

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

作　　者：邓晓敏 傅晓颖 彭佳媛 吴蔼林 李沁轩 杜旌畅 陈玮[1] 朱彦锋[1] 余小平[1] DENG Xiaomin;FU Xiaoying;PENG Jiayuan;WU Ailin;LI Qinxuan;DU Jingchang;CHEN Wei;ZHU Yanfeng;YU Xiaoping(Chengdu Medical College,Chengdu 610066,Sichuan Province,China;Department of Pathology,Leshan People’s Hospital,Leshan 614000,Sichuan Province,China;Technology Department,the Second Affiliated Hospital of Chengdu Medical College(416 Hospital of Nuclear Industry),Chengdu 610066,Sichuan Province,China)

机构地区：[1]成都医学院,四川省成都市610500 [2]乐山市人民医院病理科,四川省乐山市614000 [3]成都医学院第二附属医院(核工业四一六医院)科技部,四川省成都市610066

出　　处：《眼科新进展》2021年第2期120-124,共5页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助项目(编号81773432、81573154);四川省科技计划项目(编号2018JY0508)。

摘　　要：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5μmol·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,常规避光培养48 h;光照组,(2000±200)lux光照培养48 h;光照飞燕草素组,(2000±200)lux光照培养24 h,换含5μmol·L^(-1)飞燕草素的培养基继续光照培养24 h;避光飞燕草素组,避光培养24 h,换含5μmol·L^(-1)飞燕草素的培养基继续避光培养24 h;光照NAC组,(2000±200)lux光照培养24 h,换含3 mmol·L^(-1) NAC的培养基继续光照培养24 h。采用显微镜观察各组细胞状态、CCK-8检测细胞生存率、DCFH-DA荧光探针染色检测细胞活性氧(ROS)水平、JC-10染色检测线粒体膜电位、Annexin V-FITC/PI染色检测细胞凋亡率、Western blot检测氧化应激线粒体凋亡通路相关蛋白表达水平。结果显微镜下可见,对照组细胞生长状态良好;光照组细胞皱缩卷曲,脱落细胞增加。CCK-8检测结果显示与对照组相比,光照组细胞生存率均明显降低,差异有统计学意义(P<0.05);避光飞燕草素组细胞生存率无明显变化(P>0.05)。与光照组相比,光照飞燕草素组、光照NAC组和避光飞燕草素组细胞生存率均明显回升,差异均有统计学意义(均为P<0.05)。与对照组相比,光照组ROS含量均明显上升,差异有统计学意义(P<0.05);避光飞燕草素组ROS含量减少不明显,差异无统计学意义(P>0.05)。与光照组相比,光照飞燕草素组、光照NAC组和避光飞燕草素组ROS含量均明显下降,差异均有统计学意义(均为P<0.05)。与对照组相比,光照组线粒体膜电位均明显下降,差异有统计学意义(P<0.05);避光飞燕草素组线粒体膜电位变化不明显,差异无统计学意义(P>0.05)。与光照组相比,光照飞燕草素组、光照NAC组和避光飞燕�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 5μmol·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 followscontrol 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 5μmol·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 5μmol·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.Cell status was observed under the microscope,cell survival rate was measured by CCK-8,reactive oxygen species(ROS)level was measured by DCFH-DA fluorescent probes staining,mitochondrial membrane potential was measured by JC-10 staining,apoptosis rate was measured by Annexin V-FITC/PI staining,and the expression level of proteins related to oxidative stress mitochondrial apoptosis pathways was detected by Western blot.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 de

关 键 词：飞燕草素 661W细胞 光化学损伤 氧化应激 细胞凋亡 

分 类 号：R774[医药卫生—眼科]