660 nm红光照射对营养缺乏人神经母细胞瘤细胞活力的调节作用及其机制  

Effect of 660 nm red light on viability of human neuroblastoma cells with nutritional deficiency and its mechanism

作　　者：刘岩松 张洁 卢涛 LIU Yansong;ZHANG Jie;LU Tao(School of Life Sciences,Beijing University of Chinese Medicine,Beijing 102488,China)

机构地区：[1]北京中医药大学生命科学学院,北京102488

出　　处：《山东医药》2022年第14期36-40,共5页Shandong Medical Journal

摘　　要：目的观察660 nm红光对营养缺乏状态下人神经母细胞瘤细胞活力的调节作用,并探讨其作用机制与线粒体功能的关系。方法取人神经瘤细胞母细胞,培养于低营养培养基(RPMI1640培养基中加入终体积5%FBS和1%青—链霉素溶液),用CCK-8法验证营养缺乏模型。使用LED芯片与可调直流稳压电源制作光照装置,对营养缺乏细胞分别给予能量密度为2、4、8、16 J/cm^(2)和功率密度为2、4、8 mW/cm^(2)的660 nm红光照射,采用CCK-8试剂检测细胞活力,以细胞活力最高组对应的功率密度和能量密度作为最佳光照条件。取营养缺乏细胞,随机分为两组,光照组在最佳光照条件下接受红光照射1 h,对照组在相同环境下避光孵育1 h,运用长时程活细胞成像仪器Incucyte监测细胞增殖情况,计算光照后8、16、24 h时各组细胞平均汇合度;使用荧光探针JC-1检测光照后1~6 h线粒体膜电位和活性氧(ROS)水平。结果在功率密度2 mW/cm^(2)、能量密度8 J/cm^(2)时,光照组细胞活力最高(P均<0.01)。光照组光照后8、16、24 h的细胞汇合度与对照组比较差异均无统计学意义(P均>0.05)。光照组线粒体膜电位水平随光照时间延长逐渐升高,光照后2、3 h时达到高峰(P<0.05或<0.01),此后随着时间增加逐渐回落至光照前水平;ROS水平随光照时间延长逐渐降低,在光照后3 h达到最低(P<0.05),此后随着时间增加又出现上升(P<0.05)。结论在功率密度2 mW/cm^(2)、能量密度8 J/cm^(2)条件下,660 nm红光照射能够增加处于营养缺乏状态下的人神经母细胞瘤细胞活力;其机制可能是通过快速超极化线粒体膜电位、降低ROS生成,从而增强线粒体功能,以提高细胞活力。Objective To observe the effect of 660 nm red light on the viability of human neuroblastoma cells in a nutrient-deficient state and to investigate the relationship between its mechanism of action and mitochondrial function.Methods Human neuroma cells were taken and cultured in low nutrient medium(RPMI1640 medium with final volume of 5% FBS and 1% penicillin solution),and the nutrient-deficient model was validated by CCK-8 method.We used an LED chip with an adjustable DC regulated power supply to make a light device.The cells were irradiated with 660 nm red light at energy densities of 2,4,8 and 16 J/cm^(2) and power densities of 2,4 and 8 mW/cm^(2),respectively.Cell viability was detected by CCK-8 reagent,and we took the power density and energy density of the group with the highest cell viability as the optimal light conditions.The nutrient-deficient cells were randomly divided into two groups:the light group and the control group;the nutrient-deficient cells in the light group received red light irradiation for 1 h under the optimal light conditions,and the nutrient-deficient cells in the control group were incubated for 1 h under the same environment and protected from light.The cell proliferation was monitored using a long-range live cell imaging instrument Incucyte,and the average confluence of cells in each group was calculated at 8,16 and 24 h after light;the fluorescent probe JC-1 was used to detect the mitochondrial membrane potential from 1 to 6 h after light and reactive oxygen species(ROS)levels were measured using the fluorescent probe JC-1.Results The cell viability of the light group was highest at a power density of 2 mW/cm^(2) and an energy density of 8 J/cm^(2)(P<0.01).There were no statistically significant differences in the cell confluence between the light group and the control group at 8,16 and 24 h after illumination(all P>0.05).The mitochondrial membrane potential level in the light group gradually increased over the light time,reaching the peak at 2 and 3 h after light(P<0.05 or P<0.01),

关 键 词：人神经母细胞瘤细胞 光生物调节 660 nm红光 细胞活力 线粒体 活性氧 

分 类 号：R739.4[医药卫生—肿瘤]