CAJ | 학술논문

目的研究亚砷酸钠( sodium arsenite)对大鼠胰岛β细胞株INS-1(INS-1细胞)的凋亡作用及其机制.方法将INS-1细胞暴露于不同浓度的亚砷酸钠,首先进行噻唑蓝(MTT)试验,并用荧光分光光度计测定INS-1细胞线粒体膜电位及溶酶体膜电位的吸光度(A)值；并于亚砷酸钠作用后,应用荧光显微镜和流式细胞仪观察并检测INS-1细胞的凋亡情况.结果经亚砷酸钠作用后,INS-1细胞的活性明显下降,且细胞活性随着亚砷酸钠剂量的升高而降低；24h后,细胞线粒体膜电位荧光值明显下降,且随着亚砷酸钠剂量的增加而降低,差异有统计学意义(P＜0.01)；48h后,细胞溶酶体膜电位荧光值明显上升,且随着亚砷酸钠剂量的增加而升高,差异有统计学意义(P＜0.01)；72 h后,荧光显微镜下观察细胞,出现凋亡,随着亚砷酸钠剂量的增加,凋亡的细胞也随之增多,镜下呈亮蓝色,胞核固缩、裂解为碎块出现凋亡小体和核碎裂,部分染色质出现浓缩状态；流式细胞仪检测结果,经亚砷酸钠处理后,凋亡的细胞明显增多,且随着亚砷酸钠剂量的增高,凋亡细胞数量也随之增多.结论亚砷酸钠通过线粒体-溶酶体途径诱导大鼠胰岛β细胞株INS-1凋亡.
목적 연구아신산납( sodium arsenite)대대서이도β세포주INS-1(INS-1세포)적조망작용급기궤제.방법 장INS-1세포폭로우불동농도적아신산납,수선진행새서람(MTT)시험,병용형광분광광도계측정INS-1세포선립체막전위급용매체막전위적흡광도(A)치；병우아신산납작용후,응용형광현미경화류식세포의관찰병검측INS-1세포적조망정황.결과 경아신산납작용후,INS-1세포적활성명현하강,차세포활성수착아신산납제량적승고이강저；24h후,세포선립체막전위형광치명현하강,차수착아신산납제량적증가이강저,차이유통계학의의(P＜0.01)；48h후,세포용매체막전위형광치명현상승,차수착아신산납제량적증가이승고,차이유통계학의의(P＜0.01)；72 h후,형광현미경하관찰세포,출현조망,수착아신산납제량적증가,조망적세포야수지증다,경하정량람색,포핵고축、렬해위쇄괴출현조망소체화핵쇄렬,부분염색질출현농축상태；류식세포의검측결과,경아신산납처리후,조망적세포명현증다,차수착아신산납제량적증고,조망세포수량야수지증다.결론 아신산납통과선립체-용매체도경유도대서이도β세포주INS-1조망.
Objective To study mechanism of the apoptosis of rat pancreas islet β cell strain (INS-1cells) induced by sodium arsenite.Methods INS-I cells were exposed to sodium arsenite at the different concentrations.MTr assay was used to detect the viability of INS-1 cells.The potentials on mitochondrial membrane and lysosome membrane of INS-1 cells were determined with the fluorescence spectrophotometer.The apoptotic levels of INS-1 cells exposed to sodium arsenite were observed by a fluorescence microscope and flow cytometry.Results After exposure to sodium araenite,the viability of INS-1 cells significantly decreased with the doses of sodium arsenite.At 24 h after exposure,the OD values of the mitochondrial membrane potentials declined observably with the doses of sodium arsenite (P＜0.01).At 48 h after exposure,the OD values of the lysosome membrane potentials significantly increased with the doses of sodium arsenite (P＜0.01).At 72 h after exposure,the apoptotic cells were observed under a fluorescence microscope and enhanced with the doses of sodium arsenite.The apoptosis cells with light blue,karyopyknosis,karyorrhexis,apoptotic body and chromatin concentration appeared.The results detected with flow cytometry indicated that after exposure,the apoptotic INS-1 E cells significantly increased with the doses of sodium arsenite.Conclusions The sodium arsenite can induce the apoptosis of INS-1 cells through the mitochondria-lysosome pathway.