白藜芦醇抗黑碳颗粒对人鼻黏膜上皮细胞毒性作用及其机制

Protective effect of resveratrol on toxic effect of human nasal epithelial cells induced by black carbon

目的 探讨黑碳颗粒(BC)对人鼻黏膜上皮细胞(hNECs)的毒性作用、毒性机制以及白藜芦醇的保护作用。方法 先用不同浓度的黑碳(0、6.5、12.5、25.0、50.0、100.0μg/mL)刺激hNECs,观察BC对hNECs的毒性作用;其次,将h NECs随机分成对照组、BC组、白藜芦醇组,BC组用半致死剂量的BC浓度刺激细胞,白藜芦醇组的浓度为50μmol/L,分别检测3组细胞的存活率,细胞内ROS、SOD和CAT活性,以及8-OHdG蛋白的表达,用彗星试验检测3组细胞DNA损伤的程度。结果 BC对hNECs有细胞毒性作用,并呈剂量依赖性,BC浓度为50μg/mL时细胞存活率为(56.67±4.31)%,以此作为后续实验BC组的刺激浓度。与对照组相比,BC组细胞ROS活性升高、SOD和CAT活性下降,8-OHdG蛋白的表达上调,彗星实验中的各项指标尾长、彗星长、尾距显示DNA损伤,差异均有统计学意义(P<0.05);与BC组相比,白藜芦醇组ROS活性下降,SOD和CAT活性升高,8-OHdG蛋白的表达下调,彗星实验的指标尾长、彗星长、尾距显示DNA损伤程度有所减轻,差异均有统计学意义(P<0.05)。结论 BC对hNECs有毒性作用,可能与氧化应激有关;白藜芦醇可能通过减轻氧化应激反应对鼻黏膜上皮细胞产生保护作用。

Objective To explore the toxic effect and mechanism of black carbon(BC)on human nasal epi⁃thelial cells and the protective effect of resveratrol.Methods Human nasal epithelial cells were stimulated with dif⁃ferent concentrations of black carbon(0,6.5,12.5,25.0,50.0,100.0μg/mL)and to observe the toxic effect of black carbon on human nasal epithelial cells.Then the nasal epithelial cells were divided into control group,BC group and resveratrol protection group.The BC group was stimulated with a half lethal dose of BC,and the resvera⁃trol concentration in the protection group was 50μmol/L.In each group the survival rate of cells,the activities of ROS,SOD and CAT in cells,the expression of 8⁃OHdG protein were detected.And DNA damage were detected by comet assay.Results Black carbon particles have cytotoxicity on human nasal epithelial cells in a dose⁃dependent manner.The half lethal dose was 50μg/mL,which is used as the stimulation concentration.In BC group the activity of ROS increased,SOD and CAT decreased,and the expression of 8⁃OHdG protein increased.The tail length,com⁃et length and tail distance in comet experiment showed that DNA damaged.Compared with the control group the dif⁃ference was statistically significant(P<0.05).Compared with the BC group,ROS activity decreased,SOD activity and CAT activity increased,and the expression of 8⁃OHdG protein decreased in resveratrol protected group.The tail length,comet length and tail distance of comet experiment showed that the degree of DNA damage was reduced,and the difference were statistically significant(P<0.05).Conclusion The toxicity of BC to human nasal epithelial cells is related to oxidative stress.Resveratrol can protect nasal epithelial cells by reducing oxidative stress.