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为探究黄芪多糖免疫草鱼母本所产子代早期发育阶段体液免疫因子IgM、C3和LSZ的表达特性及代间传递效率。采用ELISA、RT-qPCR等方法分析了饲喂黄芪多糖草鱼母本血液及其子代早期发育阶段3种免疫因子的蛋白质活性及mRNA水平。结果显示,黄芪多糖免疫草鱼母本血液中IgM、C3和LSZ蛋白活性均显著高于对照组。在子代早期阶段中,3种免疫因子蛋白活性呈先下降后上升趋势,实验组IgM蛋白活性在各阶段均显著高于对照组,在卵子、5d和28d仔鱼中分别提高了2.2倍、1.7倍和1.8倍。实验组C3活性在卵子、24h胚胎分别提高了1.9倍和1.6倍。实验组LSZ活性在卵子、5d和28d仔鱼中分别提高了2.4倍、2.0倍和1.9倍;在卵子和受精卵时期3种免疫因子mRNA水平显著高于对照组。在24h胚胎至5d仔鱼没有检测到IgM和LSZ mRNA,而在14d后3种免疫因子的mRNA均呈上调表达,但Ig M和C3 mRNA水平与对照组无显著差异。GCRV攻毒后,实验组2月龄草鱼脾脏和头肾中IgM mRNA水平显著高于对照组。结果表明,黄芪多糖能够提高草鱼母本免疫能力并向子代垂直传递,在应对GCRV感染时发挥一定免疫保护作用。
To investigate the expression characteristics of humoral immune factors (IgM, C3 and LSZ) and the efficiency of inter-generational transfer in the progeny of the progeny of Astragalus polysaccharides-immunized grass carp. The protein activities and mRNA levels of three immune factors in the blood and their offspring fed with Astragalus polysaccharides (Carassius auratus polysaccharide) were analyzed by ELISA and RT-qPCR. The results showed that the activity of IgM, C3 and LSZ protein in the milk of Astragalus polysaccharide-immunized grass carp was significantly higher than that of the control group. In the early stage of offspring, the activity of three kinds of immune factors decreased first and then increased. The activity of IgM in experimental group was significantly higher than that of control group in all stages, increased by 2.2 times and 1.7 times in eggs, 5d and 28d Times and 1.8 times. The activity of C3 in experimental group increased 1.9 times and 1.6 times respectively in egg and 24h embryos. The activity of LSZ in experimental group increased by 2.4 times, 2.0 times and 1.9 times respectively in ovary, 5d and 28d larvae, and the mRNA levels of three immune factors in ovary and fertilized egg were significantly higher than those in control group. IgM and LSZ mRNA were not detected in the embryos from embryonic day 5 to day 5, while the mRNA expression of the three immune factors was up-regulated after 14 days. However, the levels of Ig M and C3 mRNA were not significantly different from those of the control group. After challenge with GCRV, the level of IgM mRNA in the spleen and head kidney of 2-month-old grass carp was significantly higher than that of the control group. The results showed that astragalus polysaccharides can improve the immune capacity of grass carp and transmitted vertically to the offspring, GCRV infection in response to a certain degree of immune protection.