为了探讨普安银鲫(Carassius auratus gibelio)卵黄囊期脂蛋白脂酶(Lipoprotein lipase,LPL)和肝脂酶(Hepatic lipase,HL)的表达特点及葡萄糖和维生素C溶液分别浸泡对它们的影响,本研究采用荧光定量PCR技术检测LPL和HL基因在普安银鲫卵黄囊仔鱼发育中的表达情况及mRNA表达水平,并检测了葡萄糖、维生素C对这两种基因mRNA表达量的影响。结果显示,LPL和HL基因在普安银鲫内源营养期、混合营养期和外源营养期均有表达,且LPL和HL mRNA表达量呈上升变化。葡萄糖组LPL和HL mRNA表达量呈上升变化,维生素C组也呈上升变化。在内源营养期、混合营养期和外源营养期,葡萄糖能显著上调LPL和HL mRNA的表达量(P〈0.05);在内源营养期、混合营养期和外源营养期,维生素C能显著上调LPL mRNA的表达量,而HL mRNA的表达量在混合营养期和外源营养期显著高于对照组(P〈0.05)。研究表明,普安银鲫发育至混合营养期时,机体内脂质分解代谢增强。适宜水平的葡萄糖、维生素C能诱导LPL和HL mRNA的表达。
In this study we investigated the expression patterns of lipoprotein lipase and hepatic lipase genes, as well as the effects of the glucose solution and the vitamin C solution on its regulation during the development of the yolk-sac larva of Carassius auratus gibelio. In the preliminary experiments, the membrane break time and hatching rates were recorded to determine the optimal concentrations of glucose and vitamin C. Solutions without(control group) and with glucose(optimal concentration 15 g/L glucose group) or vitamin C(optimal concentration 30 mg/L vitamin C group) were used for hatching, and the gene expression patterns of lipoprotein lipase and hepatic lipase were analyzed. Real-time PCR was applied to measure the mRNA level of lipoprotein lipase and hepatic lipase, and to test how glucose and vitamin C regulated the expression of two genes during the development of the yolk-sac larva. We found that genes of lipoprotein lipase and hepatic lipase were expressed at the endogenous nutrition stage, the mixed nutrition stage and the exogenous nutrition stage, and that the expression was increased over time. The mRNA levels of both lipoprotein lipase and hepatic lipase were up-regulated from the endogenous nutrition stage to the exogenous nutrition stage in the group treated with 15 g/L glucose(P〈0.05). Similarly they were also re-regulated from the endogenous nutrition stage to the exogenous nutrition stage in the group treated with 30 mg/L vitamin C, which caused a significant increase in the expression of lipoprotein lipase mRNA during the yolk-sac larva developmental stage. Vitamin C increased the expression of hepatic lipase mRNA during the mixed nutrition stage and the exogenous nutrition stage(P〈0.05). Our study suggested that the lipid catabolism of C. auratus gibelio increased during the mixed nutrition stage, and that appropriate concentrations of glucose(15 g/L) and vitamin C(30 mg/L) could induce the mRNA expression of lipoprotein lipase and hepatic lipase.