CAJ | 학술논문

通过倒置显微镜观察斜纹夜蛾细胞凋亡小体,采用MTT法测定细胞增殖抑制作用,并使用流式细胞仪检测细胞周期时相,研究了骆驼蓬碱、喜树碱、蓖麻碱、苦参碱、博落回碱和烟碱对斜纹夜蛾离体培养细胞系SL-1的作用方式和细胞毒理.结果表明:骆驼蓬碱5.0μg/mL、20.0μg/mL和喜树碱0.5μmol/L、1.0μmol/L处理后48~60 h,斜纹夜蛾离体培养细胞系SL-1表现出明显的细胞凋亡诱导作用,处理后12～48 h,明显抑制SL-1细胞增殖,流式细胞仪检测显示细胞阻滞于G2/M期;博落回碱和蓖麻碱5.0μg/mL、 20.0μg/mL对SL-1具有细胞增殖抑制作用,以20.0μg/mL处理具有细胞周期时相阻滞作用,同样使细胞阻滞于G2/M期,但无明显的细胞凋亡诱导作用;烟碱和苦参碱在5.0μg/mL、 20.0μg/mL对SL-1具有细胞增殖抑制作用.
통과도치현미경관찰사문야아세포조망소체,채용MTT법측정세포증식억제작용,병사용류식세포의검측세포주기시상,연구료락타봉감、희수감、비마감、고삼감、박락회감화연감대사문야아리체배양세포계SL-1적작용방식화세포독리.결과표명:락타봉감5.0μg/mL、20.0μg/mL화희수감0.5μmol/L、1.0μmol/L처리후48~60 h,사문야아리체배양세포계SL-1표현출명현적세포조망유도작용,처리후12～48 h,명현억제SL-1세포증식,류식세포의검측현시세포조체우G2/M기;박락회감화비마감5.0μg/mL、 20.0μg/mL대SL-1구유세포증식억제작용,이20.0μg/mL처리구유세포주기시상조체작용,동양사세포조체우G2/M기,단무명현적세포조망유도작용;연감화고삼감재5.0μg/mL、 20.0μg/mL대SL-1구유세포증식억제작용.
In order to determine and contrast the mode of action and cell toxicity mechanism of six botanical alkaloids(namely,harmaline,camptothecin,ricinine,matrine,macleay cordata and nicotine) against insect cell line,the apoptotic bodies,the inhibition of cell proliferation and the cell cycle phases were investigated by inverted phase contrast microscope,MTT test,and flow cytometry,respectively.The results showed that obvious apoptotic bodies appeared in Spodoptera litura cultured cell line SL-1 after 48～60 h treatments with harmaline at concentrations of 5.0 μg/mL and 20.0 μg/mL or camptothecin at concentrations of 0.5 μmol/L and 1.0 μmol/L.12～48 h after the same treatments,it showed obvious inhibition of cell proliferation and G2/M arrest of SL-1.It also showed that similar inhibitive effect on cell proliferation and G2/M arrest on SL-1 cell line after the treatments with macleay cordata alkaloid and ricinine at the concentrations of 5.0 μg/mL and 20.0 μg/mL respectively,but no obvious apoptosis induction.The only significant SL-1 cell line proliferation inhibition was got after the treatment with nicotine and matrine at the concentrations of 5.0 μg/mL and 20.0 μg/mL.These results provided some valuable information to the investigation of the diversity of activities of botanical alkaloids against insect pests and their cell toxicity mechanism.