| Somatostation (SS) and Growth Hormone-releasing Factor (GRF) are synthesized and secreted by hypothalamus, which could control synthesis and secretor of growth hormone of hypophysis and regulate GH concentrations in animal and human bodies.Both gene transfection of GRF expression plasmid and neutralization of endo-SS could promote the growth of animals. Intramuscular injection is extensively used in vivo, but it is limited by the low exogenous gene expression level. In this research, GRF and SS/HBsAg eukarya expression plasmids were transferred into muscle of mice, And RNA replication vector and vector ordinary muscle tissue in mice and the expression of the impact of growth compared with a view to seeking the most suitable vector containing gene dosage, the drug most effective ways to promote animal growth.First,preparation of the different doses of different vector containing the gene microspheres. Hindlimb muscle injection method used to the 14 ~ 15 g intramuscular injection in mice, respectively 0 d, 7d, 14d, 21d, 28d, 35d from the groups that the body weight of mice, the average daily gain from the different time and the cumulative weight gain Panel to analyze the effect of weight gain in mice and then the same genetic comparison of different doses, different from the same gene vector comparison. The results showed that 35 days after the injection, 50μg contained pCMV-Rep-GRF microspheres test group,100μg contained pCMV-Rep-GRF microspheres cumulative weight gain experimental group were significantly higher than 23.25% saline (P <0.01),27.34% ( P <0.01); 100μg pIRES-GRF Group cumulative weight gain was higher than 33.91% saline (P <0.01); other dose groups was not significant, high-dose group showed that weight gain good results in the low dose group.After injection 0 to 35d, contains three different doses of pCMV-Rep-HBsAg-SS microspheres average daily gain than the control group 16.46% ~ 154.55%; contains three different doses of pIRES-HBsAg-SS microspheres average, weight gain than the control group 17.02% to 109.05%. That is, in the same circumstances, RNA replication vector can be more than an ordinary vector effective expression of foreign genes.Intramuscular injection in mice microspheres containing gene, in 7 d, 21 d, 35 d, a random orbital injection site with blood from the total RNA in serum IGF-I, anti-SS, HBsAg antigen detection in the level of mRNA gene expression to analyze different dose of the different animal carrier and the immune effect of the expression level. The results show that injection of 21 d, 35d, the same dose of 100μg RNA replication vector SS mice serum antibody levels than ordinary vector high 23.76% respectively (P <0.05), 72.58% (P <0.01). Injection of 21 to 35 d, 10μg, 50μg, 100μg replicon Vector Group mouse serum HBsAb-positive rates were 100%, 50%, 100% and 10μg, 50μg, 100μg general vector of HBsAg antigen positive rate was 50% , 0,50% in the control group HBsAg antigen positive rate almost to zero. 7 d after injection, and 21 d and 35 d, pCMV-Rep-HBsAg-SS group of mice muscle expression vector SS level higher than ordinary, were pIRES-HBsAg-SS of 1.21-fold (P <0.05, n = 3 ), 1.31-fold (P <0.05, n = 3), 1.26-fold (P <0.01, n = 3). RNA replication vector and vector ordinary mice immune to the effects of comparison, the same dose of circumstances, RNA replication vector stimulation of IGF-I in serum higher than normal vector.Second, useing of the thermo-sensitive sustained-release drug system, prepared GHRP-6 thermo-sensitive drug delivery system, and will examine its impact on the growth of the animal. The results showed that Vc Rex rabbits were injected GHRP-6 thermo-sensitive release gel 7 d, 14d, 21d, 28d, 2 g / L, 4 g / L, respectively, the cumulative weight gain was higher than 25.83% saline ~ 63.37% . Vc Rex rabbits were injected GHRP-6 thermo-sensitive different time release gel serum IGF-I concentrations: 21 d after injection, 2 g / L, 4 g / L, respectively, over 37.43% normal saline group (p <0.01) 47.17% (p <0.01); 35 d after injection, 2 g / L, 4 g / L, respectively, over 31.71% normal saline group (p <0.01), 40.14% (p <0.01).This study uses the efficient expression of the exogenous gene RNA replication vector dose and different gene to improve GRF and HBsAg / SS expression plasmid muscle in the best expression of the results show that the method in promoting animal growth has played a positive role ,but reports and related RNA replication vector ordinary vector expression level of 2-4 times there are gaps; This study also employs GHRP thermo-sensitive delivery system to examine its role in the growth of animals. After comparing experiment can be used in future experiments can be efficiently exogenous gene expression vector containing drugs with the appropriate channels, can be improving animal production performance of practical technology.
|
PDF Full Text Request