将斑马鱼饲养在持续充气的自来水中，水温保持在25？C~27？C，取15条健康斑马鱼，用108 cells/ml的嗜水气单胞菌溶液浸泡处理24 h，3 h、6 h、12 h和24 h各取3条斑马鱼以实时定量PCR技术检测补体基因的表达水平。A.h处理后，斑马鱼体内C1、C4、C6和MASP的表达水平逐步升高，而C3、Bf和MBL的表达水平则先降低再升高，但它们都在处理6 h或12 h时达到峰值，之后其表达水平逐步降低。这表明鱼类补体系统能在短时间内对外界刺激做出应答，但是由于不同补体成分及补体激活途径在机体免疫中的作用差异，它们对外界感染做出的反应也有所不同。
The healthy zebrafish were maintained in well-aerated tap water at 25？C~27？C; 15 healthy zebrafish were selected and then immersed in 20 μg/ml Aeromonas hydrophila solution for 24 h; 3 fish were sampled at 3 h, 6 h, 12 h and 24 h; the expression levels of main complement genes were analyzed by real-time fluorescent quantitative PCR. It was shown that the expression of C1r/s, C4, C6 and MASP all up-regulated after A.h treatment and reached the peak at 6 h which was followed by a decline. In contrast, down-regulation of C3, MBL and Bf expression was detected shortly after treatment, and then up-regulation followed and also peaked at 6 h or 12 h. These results indicated that complement system can effectively respond to infection in a short period of time, and different genes respond to the external stimulation in different ways, which might be caused by the different roles of complement components and complement pathways.
Wang, Z.P., Zhang, S.C. and Wang, G.F. (2008) Response of Comple-ment Expression to Challenge with Lipolopysaccharide in Embryos/larvae of Zebrafish (Daniorerio): Maturation of Complement Immunocompetence. Fish and Shellfish Immunology, 25, 264-270. https://doi.org/10.1016/j.fsi.2008.05.010
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Tang, R., Dodd, A.W., Lai, D., et al. (2007) Validation of Zebrafish (Daniorerio) Reference Genes for Quantitative Real-Time RT-PCR Normalization. Acta Biochimica et Biophysica Sinica (Shanghai), 39, 384-390.
https://doi.org/10.1111/j.1745-7270.2007.00283.x
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Livak, K.J. and Schmittgen, T.D. (2001) Analysis of Rel-ative Gene Expression Data Using Real-Time Quantitative PCR and the 2(–？？Ct) Method. Methods, 25, 402-408. https://doi.org/10.1006/meth.2001.1262
