重组生长肌动素的制备纯化及其对罗非鱼肝细胞胰岛素样生长因子基因表达的影响

Jianpeng Peng, A. Lian, Q. Jiang
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引用次数: 6

摘要

生长激素(growth hormone, GH)/催乳素(prolactin, PRL)家族成员之一的生长激素(Somatolactin, SL)是鱼类特有的具有多种功能的垂体激素。然而,对其在鱼肝细胞中的生物学功能知之甚少。以罗非鱼为模型,sl转录本在各种头外组织中广泛表达,在肝脏中表达量较高。为了探索SL在肝细胞中的生物学作用,我们制备并纯化了能诱导罗非鱼黑色素细胞色素聚集的重组罗非鱼SL蛋白。进一步制备了SL的抗血清,并通过抗血清预吸收证实了其特异性。在饥饿4周期间,从饥饿第1周开始至第4周,饥饿鱼的肝脏SL转录本显著高于对照鱼。再饲喂后,SL转录本水平恢复正常。利用罗非鱼肝细胞原代培养物,重组罗非鱼SL静态孵育可提高胰岛素样生长因子(IGF1和IGF2)基因表达,而利用SL抗血清免疫中和去除内源性SL可抑制IGF1和igf基因表达。综上所述,这些发现首次提供了证据,证明SL可能是一种新的调节剂,可以刺激鱼类肝细胞中IGF1和IGF基因的表达。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Production and Purification of Recombinant Somatolactin and its Effects on Insulin-like Growth Factors Gene Expression in Tilapia Hepatocytes
Somatolactin (SL), the member of the growth hormone (GH)/prolactin (PRL) family, is fish-specific pituitary hormone with diverse functions. However, little is known about its biological function in fish hepatocytes. Using tilapia as a model, SLtranscripts were shown to be widely expressed in various extrapituitary tissues with the relatively high expression level in liver. To explore the biological action of SL in hepatocytes, we produced and purified recombinant tilapia SL protein which could induce pigment aggregation in tilapia melanophores. Further, the antiserum for the SL was produced and its specificity was confirmed by antiserum preabsorption. During 4 week starvation, hepatic SL transcripts in starved fish were significantly higher than the control fish starting on the 1st week of starvation until 4th week. After re-feeding, the SL transcripts level returned to normal. Using primary cultures of tilapia hepatocytes, insulin-like growth factors (IGF1 and IGF2) gene expression were elevated by static incubation with recombinant tilapia SL. In contrast, removal endogenous SL by immunoneutralization using SL antiserum was shown to inhibit IGF1 and IGFgene expression. These findings, taken together, provide evidence for the first time that SL may serve as a novel regulator in fish stimulating IGF1 and IGF gene expression in hepatocytes.
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