摘要
目的研究石菖蒲活性成分β-细辛醚对过氧化氢诱导PC12细胞焦亡的保护作用及其与氧化应激的相关性。方法以不同浓度过氧化氢作用于PC12细胞,采用噻唑蓝(MTT)比色法检测其对细胞存活率影响并用蛋白免疫印迹法(western blotting)检测不同浓度过氧化氢对细胞焦亡蛋白Caspase-1表达影响,确定激活细胞焦亡最佳造模浓度;以3.75,7.5,15,30,60,120μg·ml-1浓度的β-细辛醚处理PC12细胞,采用噻唑蓝(MTT)比色法检测其对细胞活性的影响。实验随机分组为空白组,过氧化氢组,β-细辛醚低、中、高(15,30,60μg·ml-1)浓度组,CCK-8法检测β-细辛醚对细胞存活率的影响,检测各组细胞上清中LDH,细胞中CAT及GSH含量,蛋白免疫印迹法检测β-细辛醚对Caspase-1及下游IL-18表达的影响。结果与空白组细胞相比较,过氧化氢组细胞存活率显著下降(P<0.05),细胞上清液中LDH活力显著增高,细胞中CAT和GSH活力显著降低(P<0.05),Caspase-1、IL-18灰度值显著增高(P<0.05);与过氧化氢组相比,β-细辛醚预保护显著提高PC12细胞存活率(P<0.01),降低细胞上清液中LDH活力,提高细胞中CAT和GSH活力(P<0.05),显著降低Caspase-1、IL-18灰度值。结论过氧化氢可产生氧化应激并诱导PC12细胞焦亡,一定浓度β-细辛醚可抑制PC12细胞焦亡,发挥保护作用,其机制可能与抗氧化有关。
Objective To study the protective effect ofβ-asarone,an active component of Acorus tatarinowii,on the pyroptosis of PC12 cells induced by hydrogen peroxide and its relationship with oxidative stress.Methods Different concentrations of hydrogen peroxide were used to treat PC12 cells,and the effect of cell viability was determined by MTT.The effect of Caspase-1 expression was detected byWestern blot.PC12 cells were treated withβ-asarone at concentrations of 3.75,7.5,15,30,60,120μg·ml-1,the effect of cell viability was determined by MTT.The experiments were randomly divided into blank group,model group,β-asarone low,medium and high(15,30,60μg·ml-1)concentration group.The effect of cell viability was determined by CCK-8.LDH contents in the supernatant,CAT and GSH contents in the cells were detected,The effect ofβ-asarone on the expression of Caspase-1 and downstream IL-18 was detected by western blotting.Results Compared with the blank group,the survival rate of the model group was significantly decreased(P<0.05).The LDH activity in the supernatant of the model group was significantly increased,and the CAT and GSH activities in cells were significantly decreased(P<0.05).The gray value of Caspase-1 and IL-18 was significantly increased(P<0.05).Compared with the model group,β-asarone pre-protection significantly increased the survival rate of PC12 cells(P<0.01),and significantly decreased the activity of LDH,increased the activity of CAT and GSH in cells(P<0.05),and significantly decreased the gray value of Caspase-1 and IL-18.Conclusion Hydrogen peroxide can produce oxidative stress and induce pyroptosis of PC12 cells.β-asarone can effectively inhibit pyroptosis of PC12 cells and play a protective role.
作者
黑鑫鑫
刘涛
HEI Xin-xin;LIU Tao(School of Basic Medicine,Nanjing University of Chinese Medicine,Nanjing,Jiangsu 210023,China)
出处
《时珍国医国药》
CAS
CSCD
北大核心
2019年第5期1025-1028,共4页
Lishizhen Medicine and Materia Medica Research
基金
国家自然科学基金(81173313)
