摘要
绿僵菌素A和B是生物活性分子,本研究用MTT法、相差显微观察、荧光倒置显微观察和流式细胞术比较了绿僵菌素A和B对斜纹夜蛾Spodoptera litura SL-1细胞的毒杀作用。结果表明:绿僵菌素A和B对SL-1细胞均具有明显的增殖抑制作用,且具有正比的时间-浓度-效应关系,绿僵菌素A和B处理细胞48h后的IC50值分别为7.80和20.73μg/mL。倒置相差显微观察发现,绿僵菌素A和B可以引起SL-1细胞变圆、胞膜收缩、有凋亡小颗粒形成。随着时间的延长,细胞悬浮致死并出现大量空泡和胞质外泄现象。但是在同样的处理浓度下,绿僵菌素A的作用较绿僵菌素B明显。用AO/EB染色后,荧光显微观察发现:绿僵菌素A诱导的细胞荧光强度高于绿僵菌素B。流式细胞仪检测结果表明:绿僵菌素A和B对SL-1细胞具有明显的致凋亡作用。10μg/mL绿僵菌素A和B作用细胞48h后,SL-1细胞的总凋亡率分别达78.88%±0.97%和72.23%±2.29%。本研究从细胞水平肯定了绿僵菌素具有良好的增殖抑制和致凋亡作用,并且为它在害虫防治中的潜在应用提供了一些理论支持。
Both destruxin A and destruxin B are bioactive molecules.In the present study,the effects of the two destruxins（destruxin A and destruxin B） on Spodoptera litura SL-1 cells were compared with MTT assay,inverted phase contrast microscope（IPCM）,fluorescence microscopy（FM） and flow cytometry（FCM） technologies.The results showed that both destruxins had an apparent effect on inhibition of cellular proliferation,and they had a positive relationship of time-concentration-efficacy.The IC50 values were 7.80 and 20.73 μg/mL at 48 h after treatment with destruxin A and destruxin B,respectively.With inverted phase contrast microscopy observation,we found that both destruxins could cause cell rounded,celluar membrane shrunk,and apoptotic bodies formed.Most of cells were suspended and dead as vacuoles appeared and cytoplasm leaked out with extension of the treatment time.However,under the same concentrations tested,destruxin A showed stronger effect than destruxin B.With fluorescence microscopy observation after AO/EB staining,a relative higher fluorescent intensity of the cells induced by destruxin A was found as compared with destruxin B.Observations by flow cytometer showed that destruxin A and destruxin B could induce apoptosis in SL-1 cells,and the total apoptotic rates reached 78.88% ±0.97% and 72.23% ±2.29% at 48 h after treatment with 10 μg/mL destruxin A and destruxin B,respectively.This research demonstrated that destruxins possess a preferable activity for cell proliferation and apoptosis at the cellular level,and provided some theoretical support for their potential application in pest control.
出处
《昆虫学报》
CAS
CSCD
北大核心
2011年第9期1003-1009,共7页
Acta Entomologica Sinica
基金
国家自然科学基金项目(31071685
30800718)
