摘要
目的研究鱼藤素致人支气管上皮细胞(16HBE)毒性与线粒体损伤和凋亡的相关性,为研究鱼藤素毒性提供参考。方法将1.56~100μmol·L^(-1)鱼藤素与16HBE细胞共孵育,培养24~72 h,CCK-8法测定细胞抑制率,得到半数抑制浓度(IC 50),确定鱼藤素中剂量。将16HBE细胞分为对照组(正常培养),鱼藤素小剂量组(15μmol·L^(-1)鱼藤素),鱼藤素中剂量组(30μmol·L^(-1)鱼藤素)及鱼藤素大剂量组(60μmol·L^(-1)鱼藤素),培养24 h,倒置荧光显微镜和透射电镜观察线粒体和细胞核超微结构形态,比色法测定谷胱甘肽过氧化物酶(GPx)和超氧化物歧化酶(SOD)活力,流式细胞术检测线粒体膜电位(MMP)和细胞凋亡率。结果鱼藤素对16HBE增殖具有抑制作用,且抑制率呈浓度和时间依赖关系(P<0.05),24,48和72 h IC 50分别为(32.95±2.39),(21.07±2.21)和(15.46±0.93)μmol·L^(-1)。与对照组比较,鱼藤素作用后细胞核凋亡,线粒体明显肿胀、空泡化和嵴脱落,GPx与SOD活力显著下降,MMP急剧下降;对照组荧光强度(57682±719.1),鱼藤素小剂量组、中剂量组、大剂量组分别为(51593±1525.4),(29427±578.4)和(23575±1366.8);鱼藤素各剂量组细胞凋亡率明显上升,且呈浓度依赖性(P<0.05)。结论鱼藤素致16HBE细胞毒性可能与线粒体损伤和细胞凋亡相关。
Objective To investigate the relevance of mitochondrial damage,cell apoptosis and deguelin-induced cytotoxicity in 16HBE cells,and to provide reference for the research about deguelin-induced toxicity.Methods The 16HBE cells were treated with 1.56-100μmol·L^(-1)deguelin for 24-72 h.CCK-8 assay was used to detect the inhibitory effect of deguelin on proliferation of 16HBE cells.The half inhibition concentration(IC 50)values of deguelin on 16HBE cells were calculated to determine the dose of medium-concentration group.The cells were divided into control group(normally cultured 16HBE cells),deguelin low-concentration group(15μmol·L^(-1)deguelin),medium-concentration group(30μmol·L^(-1)deguelin)and high-concentration group(60μmol·L^(-1)deguelin).After treated with different dose of deguelin for 24 hours,16HBE cells were observed under inverted microscope and transmission electron microscope to detect their morphological changes of mitochondria and cell nucleus.GPx and SOD activity were determined by assay kits according the manufacturer's protocol.Flow cytometry was used to detect the levels of MMP and apoptosis.Results Compared with the control group,deguelin can inhibit the proliferation rate of 16HBE cells in concentration-dependent and time-dependent manner(P<0.05).IC 50 for 24,48 and 72 h were(32.95±2.39),(21.07±2.21)and(15.46±0.93)μmol·L^(-1),respectively.Compared with the control group,nuclear apoptosis can be observed.Mitochondrial swelling,cristae disruption and vacuolar degeneration can be observed obviously.GPx activity,SOD activity and the levels of MMP decreased significantly(P<0.05).The fluorescence intensity in control group,low-concentration group,medium-concentration group and high-concentration group were(57682±719.1),(51593±1525.4),(29427±578.4)and(23575±1366.8),respectively.What's more,cell apoptosis in treated groups increased significantly.Conclusion There may exist a relevance between mitochondrial damage,cell apoptosis and deguelin-induced cytotoxicity of 16HBE cells.
作者
方琼彤
吕满霞
吴新荣
FANG Qiongtong;LYU Manxia;WU Xinrong(Department of Pharmacy,the Fifth Affiliated Hospital of SUNYAT-SEN University,Zhuhai 519000,China;Department of Pharmacy,General Hospital of Southern Theatre Command of PLA,Guangzhou 510010,China;China-Singapore Joint Institute of International Studies,Guangzhou 510700,China)
出处
《医药导报》
CAS
北大核心
2021年第7期870-875,共6页
Herald of Medicine
基金
全军后勤科研计划(No CGZ14C003)
国家自然科学基金资助项目(No 81673608)。
