title: Reconstituted High-Density Lipoprotein Containing Human Growth Hormone-1 Shows Potent Tissue Regeneration Activity with Enhancement of Anti-Oxidant and Anti-Atherosclerotic Activities.
abstract: BACKGROUND
  Human growth hormone-1 (GH-1), somatotropin, is a peptide hormone that stimulates cell division in tissues such as bone and cartilage.
  
  
  METHODS
  To compare physiological activities in lipid-free and lipid-bound states, we expressed and incorporated GH-1 in reconstituted high-density lipoprotein (rHDL).
  
  
  RESULTS
  GH-1 was expressed and purified using the pET30(a) vector and an Escherichia coli expression system. Purified GH-1 (at least 98% purity, 23 kD) was characterized and synthesized with apolipoproteinA-I (apoA-I), 1-palmitoyl-2-oleoyl-phosphatidylcholine (POPC), and cholesterol. Secondary structure analysis of GH-1 revealed 54% α-helical content in a lipid-free state and 65% α-helical content in a lipid-bound state along with blue-shifted tryptophan movement (around 2 nm). GH-1 caused less uptake of oxidized low-density lipoprotein (oxLDL) into macrophages and inhibited senescence of dermal cells in a dose-dependent manner. GH-1 in a lipid-bound state exerted stronger inhibitory activity than in a lipid-free state, indicating improved anti-atherosclerotic activity due to the lipid formulation. In a fin regeneration experiment using zebrafish (17 weeks old, n=9), GH-1 showed its highest regeneration speed without any side effects. GH-1-rHDL containing apoA-I showed 2.3-fold and 1.6-fold higher regeneration speeds than lipid-free GH-1 (in native state) and lipid-bound GH-1, respectively.
  
  
  CONCLUSIONS
  Incorporation of GH-1 and apoA-I in HDL enhanced tissue regeneration activity of amputated tail fin, indicating a synergetic effect between GH-1 and apoA-I in a lipid-bound state.
authors.affiliation: 1 School of Biotechnology, Yeungnam University , Gyeongsan, Republic of Korea.|2 Research Institute of Protein Sensor, Yeungnam University , Gyeongsan, Republic of Korea.|3 BK21plus Program Serum Biomedical Research and Education Team, Yeungnam University , Gyeongsan, Republic of Korea.
authors.forename: Eun-Young
authors.initials: EY
authors.lastname: Lee
authors.affiliation: 1 School of Biotechnology, Yeungnam University , Gyeongsan, Republic of Korea.|2 Research Institute of Protein Sensor, Yeungnam University , Gyeongsan, Republic of Korea.|3 BK21plus Program Serum Biomedical Research and Education Team, Yeungnam University , Gyeongsan, Republic of Korea.
authors.forename: So-Hee
authors.initials: SH
authors.lastname: Kim
authors.affiliation: 1 School of Biotechnology, Yeungnam University , Gyeongsan, Republic of Korea.|2 Research Institute of Protein Sensor, Yeungnam University , Gyeongsan, Republic of Korea.|3 BK21plus Program Serum Biomedical Research and Education Team, Yeungnam University , Gyeongsan, Republic of Korea.
authors.forename: Kyung-Hyun
authors.initials: KH
authors.lastname: Cho
mesh_terms: D058500:Animal Fins / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D016632:Apolipoprotein A-I / Q000494:pharmacology; D050197:Atherosclerosis / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D015153:Blotting, Western; D016922:Cellular Senescence / Q000187:drug effects*; D002942:Circular Dichroism; D004625:Embryo, Nonmammalian / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D019382:Human Growth Hormone / Q000494:pharmacology*; D006801:Humans; D008075:Lipoproteins, HDL / Q000737:chemistry*; D008264:Macrophages / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D012038:Regeneration / Q000187:drug effects*; D012867:Skin / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D015027:Zebrafish / Q000254:growth & development* / Q000378:metabolism
chemical_list: D000975:Antioxidants; D016632:Apolipoprotein A-I; D008075:Lipoproteins, HDL; C035392:high density lipoprotein-1; D019382:Human Growth Hormone