Dai Jian-wei, Liu Song-cai, Hao Lin-lin, Zhang Yong-liang, Zhang Qianqian, Ren Xiao-hui, Jiang Qing-yan
Department of Biochemistry, Veterinary College, Jinlin University, Changchun, People's Republic of China.
Anim Biotechnol. 2008;19(2):104-16. doi: 10.1080/10495390801888328.
Somatostatin (SS) and growth hormone-releasing hormone (GHRH) are synthesized and secreted by the hypothalamus, which can control the synthesis and secretion of the growth hormone (GH) from the hypophysis as well as regulate the GH concentrations in animals and humans. In this article, we describe the regulation of animal growth using plasmid DNA encoding both the GHRH gene and the SS gene fused with the hepatitis B surface antigen (HBsAg) gene. We constructed a series of expression plasmids to express the GHRH and HBsAg-SS fusion genes individually as well as collectively. The fusion gene and GHRH were successfully expressed in Chinese hamster ovary (CHO) cells, as proven by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunoblotting tests. Poly D, L-lactide-co-glycolic acid (PLGA) plasmid-encapsulating microspheres were prepared and injected intramuscularly into the leg skeletal muscles of rabbits. Weight gain/day and the levels of insulinlike growth factor-I (IGF-I), SS, and hepatitis B surface antibody (HBsAb) were monitored. During days 30 postinjection, increase in weight gain/day and IGF- I concentration and decrease in SS were observed in treatment groups. From days 15 to 30 postinjection, the weight gain/day significantly increased (P < 0.05) by 129.13%, 106.8%, and 72.82% relative to the control group in the co-expression GHRH and fusion gene (named P-G-HS), fusion gene (named P-HS), and GHRH (named P-G) groups, respectively. And most importantly, the P-G-HS group showed significant weight gain/day (P < 0.05) relative to the P-G and P-HS groups. A significant increase in the IGF-I concentration and decrease in the SS level relative to the control group were also observed. The results indicated that the combination of plasmid-mediated GHRH supplementation and positive immunization against SS led to more robust weight gain/day in rabbits.
生长抑素(SS)和生长激素释放激素(GHRH)由下丘脑合成并分泌,它们可控制垂体生长激素(GH)的合成与分泌,同时调节动物和人体内的GH浓度。在本文中,我们描述了使用编码GHRH基因和与乙肝表面抗原(HBsAg)基因融合的SS基因的质粒DNA对动物生长的调控。我们构建了一系列表达质粒,以单独或共同表达GHRH和HBsAg-SS融合基因。通过逆转录-聚合酶链反应(RT-PCR)和免疫印迹试验证明,融合基因和GHRH在中国仓鼠卵巢(CHO)细胞中成功表达。制备了聚D,L-丙交酯-乙交酯(PLGA)包裹质粒的微球,并将其肌肉注射到兔腿部骨骼肌中。监测每日体重增加量以及胰岛素样生长因子-I(IGF-I)、SS和乙肝表面抗体(HBsAb)的水平。在注射后30天内,治疗组观察到每日体重增加量增加,IGF-I浓度升高,SS降低。在注射后15至30天,与对照组相比,共表达GHRH和融合基因(命名为P-G-HS)、融合基因(命名为P-HS)和GHRH(命名为P-G)组的每日体重增加量分别显著增加(P < 0.05),增幅分别为129.13%、106.8%和72.82%。最重要的是,与P-G组和P-HS组相比,P-G-HS组的每日体重增加量显著增加(P < 0.05)。相对于对照组,IGF-I浓度也显著升高,SS水平降低。结果表明,质粒介导的GHRH补充与针对SS的阳性免疫相结合可使兔每日体重增加更为显著。