Abstract
In this study, we utilized the HIV protein Tat protein transduction domain (Tat-PTD) to enhance the intestinal absorption of recombinant human growth hormone (rhGH) delivered by oral administration. Insulin-like growth factor 1 (IGF-1), the key factor in the GH signal transduction pathway, was differentially affected at the mRNA level by various concentrations of rhGH. At high rhGH concentrations (500 ng/ml), IGF-1 was downregulated, while low concentrations (5 ng/ml) caused IGF-1 upregulation. The addition of Tat-PTD had a significant facilitating effect on rhGH. Frozen tissue sections visualized with fluorescence microscopy, cultured cells visualized by confocal microscopy and flow cytometry all confirmed that rhGH fused to Tat-PTD demonstrated more intracellular fluorescent signal when compared to rhGH alone. ELISA showed that after 2 h of incubation, Tat-rhGH levels in the rat intestinal cavity were 1.38-fold higher than rhGH. These data indicated that Tat-PTD effectively improved the internalization of rhGH and enhance the signal transduction of rhGH, possibly laying a solid foundation for the novel oral administration of rhGH.
Similar content being viewed by others
References
C.H. Li, Human growth hormone: 1974–1981. Mol. Cell. Biochem. 46, 31–41 (1982)
C.H. Li, H. Papkoff, Preparation of properties of growth hormone from human and monkey pituitary glands. Science 124(3235), 1293–1294 (1956)
Maybe, G. Nancy, Direct expression of human growth in Escherichia coli with the lipoprotein promoter, in Recombinant DNA products: insulin, interferon, and growth hormone, ed. by P.B. Arthur (CRC Press, Boca Raton, 1984)
S. Turker, E. Onur, Y. Ozer, Nasal route and drug delivery systems. Pharm. World Sci. 26, 137–142 (2004)
G. Levin, A. Gershonowitz, H. Sacks, Transdermal delivery of human growth hormone through RF-microchannels. Pharm. Res. 22, 550–555 (2005)
R.U. Agu, M.I. Ugwoke, M. Armand, R. Kinget, N. Verbeke, The lung as a route for systemic delivery of therapeutic proteins and peptides. Respir. Res. 2, 198–209 (2001)
A. Leone-Bay, K. Ho, R. Agarwal, 4-[4-[(2-Hydroxybenzoyl)amino]phenyl] butyric acid as a novel oral delivery agent for recombinant human growth hormone. J. Med. Chem. 39(13), 2571–2578 (1996)
H. Harada, M. Hiraoka, Protein transduction domain-mediated delivery of anticancer proteins. Pharm. Perspect. Cancer. Ther. 297–319 (2009)
S.R. Schwarze, A. Ho, A. Vocero-Akbani, S.F. Dowdy, In vivo protein transduction: delivery of a biologically active protein into the mouse. Science 285, 1569–1572 (1999)
T. Maniatis, E.F. Fritsch, J. Sambrook, Molecular cloning: a laboratory manual 202–203 (Cold Spring Harbor Laboratory, New York, 1982)
Y. Zhang, J. Ning, X. Qu, X. Meng, J. Xu, TAT-mediated oral subunit vaccine against white spot syndrome virus in crayfish. J. Virol. Methods 181, 59–67 (2012)
Q. Fan, C. Wu, L. Li, Some features of intestinal absorption of intact fibrinolytic enzyme ш-1 from Lumbricus rubellus. Biochim. Biophys. Acta 1526, 286–292 (2001)
S. Wu, J.R. Robinson, Transcellular and lipophilic complex-enhanced intestinal absorption of human growth hormone. Pharm. Res. 1999, 1266–1272 (1999)
H.J. Lee, G. Riley, O. Johnson, In vivo characterization of sustained-release formulations of human growth hormone. J. Pharmacol. Exp. Ther. 281, 1431–1439 (1997)
N. Amet, W. Wang, W. Shen, Human growth hormone-transferrin fusion protein for oral delivery in hypophysectomized rats. J. Control. Release 25, 177–182 (2010)
E.C. Snow, Insulin and growth hormone function as minor growth factors that potentiate lymphocyte activation. J. Immunol. 135, 776–778 (1985)
A. Gertler, A. Walker, H.G. Friesen, Enhancement of human growth hormone-stimulated mitogenesis of Nb2 nod lymphoma cells by 12-o-tetradecanoyl-phorbol-13-acetate. Endocrinology 116, 1636 (1985)
L. Matera, M. Cutufia, M. Geuna, Prolactin is an autocrine growth factor for the Jurkat human T-leukemic cell line. J. Neuroimmunol. 79, 12–21 (1997)
P.E. Mullis, T. Lund, M.S. Patel, C.G.D. Brook, Regulation of human growth hormone receptor gene expression by human growth hormone in a human hepatoma cell line. Mol. Cell. Endocrinol. 76, 125–133 (1991)
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Ni, Y., Yu, J., Xu, JP. et al. Enhanced delivery of human growth hormone across cell membrane by Tat-PTD. Endocrine 46, 138–147 (2014). https://doi.org/10.1007/s12020-013-0039-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12020-013-0039-3