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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。
Metasorubicin A and B were biologically active molecules. In this study, the effects of desmogenin A and B against Spodoptera litura SL-1 were compared by MTT assay, phase contrast microscopy, fluorescence inverted microscopy and flow cytometry. 1 cells poisoning effect. The results showed that both Metarigosin A and B had obvious proliferation inhibitory effects on SL-1 cells, and had a proportional time-concentration-effect relationship. IC50 values of cells treated with destrucstatin A and B for 48 h were 7.80 and 20.73 μg / mL. Inverted phase contrast microscopy found that the Metarigosaccharides A and B can cause SL-1 cells rounded, membrane contraction, the formation of small apoptotic particles. With the extension of time, the cells died of cell suspension and appeared a large number of vacuoles and cytoplasmic excretion phenomenon. However, the effect of Metarhizin A was more obvious than that of Metarigin B at the same treatment concentration. After staining with AO / EB, the fluorescence microscopic observation showed that the fluorescence intensity of the cells induced by desarmarin A was higher than that of the strain. Flow cytometry results show that: Metarigosin A and B on SL-1 cells have a significant role in apoptosis. The total apoptotic rate of SL-1 cells reached 78.88% ± 0.97% and 72.23% ± 2.29% respectively after treated with 10μg / mL SC. This study affirmed that SCs have a good effect of inhibiting proliferation and inducing apoptosis at the cellular level and provide some theoretical support for its potential application in pest control.