| The silkworm, Bombyx mori, is an important economic insect, is a typical patternof insects. Targeted research on apoptosis of Bombyx mori has been reported, forexample, the blood of B. mori can play the role of anti-apoptotic effects, the moltinghormone can induce cell apoptosis. But compared with the Nematode, Drosophila,human and other model organisms, the study on the apoptosis pathway in B. mori is stillat the starting stage. The study on apoptosis in silkworm ovarian cell lines (BmN) israrely reported. In this study, at the background of induction factors (H2O2, virus) invitro, we do the research on the cell morphology of apoptosis and gene expression leveldifferences by technologies like morphological observation, agarose gel electrophoresisand quantitative real-time PCR. We also explore the role of two kinds of ace of B. moriin the process of apoptosis, the results are as follows.Apoptosis was induced by H2O2.BmN cells were induced by H2O2at0.100mol/L for3h, the features of apoptosis can be observed in the optical and fluorescencemicroscopy, ladder shape strap can be obviously observed by agarose gelelectrophoresis after24h induction. The QRT-PCR results showed that, the expressionof Bombyx mori, telomerase gene, Bmtrt decreased; the expression of apoptosis-relatedgenes of B. mori Bmcaspase-5gradually increased. Compared with the control group,41.942times increased at24h; the expression of Bmp53gene decreased at first and thengradually increased and1.592times increased at24h. The results showed that afterH2O2induction, the B. mori apoptosis-related genes have emerged characteristicexpression, the relationship between genes in the process of apoptosis will be studied inthe future.Apoptosis was induced by Bombyx mori nuclear polyhedrosis virus, BmNPV.After the infection by BmNPV solution30μL (1×108PFU/mL) virus for24h, BmN cell became round, smooth edged; the characteristics of apoptosis are showed obviouslyafter48h. The QRT-PCR results showed that, apoptosis-related genes have significantchanges in transcript levels. The expression of Bmcaspase-1and Bmice-5geneincreased significantly at48h,15.195times,6.245times increased respectively; theexpression of Bmcaspase-5and Bmp53gene up-regulated significantly at72h,22.911times,4.610times increased respectively; the expression of Bmtrt decreased. Theresults showed that apoptosis occurs in BmN cells after infection by BmNPV, whichcould provide new reference information for the study of BmNPV infectionmechanisms.The expression characteristics acetylcholinesterase genes of B. mori duringapoptosis. We studied the expression characteristics of ace genes in B. mori afterinduction by H2O2and BmNPV. The results showed that the expression of ace1geneincreased gradually, but the expression of ace2decreased during the process ofapoptosis H2O2-induced; the expression of ace1and ace2gene increased in varyingdegrees, the expression of ace1gene is more obvious. No matter in normal cells orapoptotic cells, the expression level of ace2gene are almost higher than that of ace1gene expression, the maximum of ace2/ace1is up to67.449. The results show that, ace1gene play an important role in the apoptotic process of BmN cells; and ace2gene mayhave different functions after induction by different inducers, its mechanism needsfurther study.The results of this study provide a reference to explore the apoptotic mechanism ofB. mori cells under different conditions. |
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