In ovo administration of human recombinant leptin shows dose dependent angiogenic effect on chicken chorioallantoic membrane

BACKGROUND: Leptin, the cytokine produced by white adipose tissue is known to regulate food energy homeostasis through its hypothalamic receptor. In vitro studies have demonstrated that leptin plays a major role in angiogenesis through binding to the receptor Ob-R present on ECs by stimulating and i...

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Autores principales:	Manjunathan,Reji, Ragunathan,Malathi
Lenguaje:	English
Publicado:	Sociedad de Biología de Chile 2015
Materias:	Angiogenesis HRL Human Recombinant Leptin CAM chorioallantoic membrane ECs endothelial cells VEGF vascular endothelial growth factor MMP matrix metalloproteinase Blood vessels CD34
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602015000100029
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spelling	oai:scielo:S0716-976020150001000292016-02-16In ovo administration of human recombinant leptin shows dose dependent angiogenic effect on chicken chorioallantoic membraneManjunathan,RejiRagunathan,Malathi Angiogenesis HRL Human Recombinant Leptin CAM chorioallantoic membrane ECs endothelial cells VEGF vascular endothelial growth factor MMP matrix metalloproteinase Blood vessels CD34 BACKGROUND: Leptin, the cytokine produced by white adipose tissue is known to regulate food energy homeostasis through its hypothalamic receptor. In vitro studies have demonstrated that leptin plays a major role in angiogenesis through binding to the receptor Ob-R present on ECs by stimulating and initiating new capillary like structures from ECs. Various in vivo studies indicate that leptin has diverse effect on angiogenesis. A few reports have showed that leptin exerts pro angiogenic effects while some suggested that it has antiangiogenic potential. It is theoretically highly important to understand the effect of leptin on angiogenesis to use as a therapeutic molecule in various angiogenesis related pathological conditions. Chicken chorio allantoic membrane (CAM) on 9th day of incubation was incubated with 1, 3 and 5 μg concentration of HRL for 72 h using gelatin sponge. Images where taken after every 24 h of incubation and analysed with Angioguant software. The treated area was observed under microscope and histological evaluation was performed for the same. Tissue thickness was calculated morphometrically from haematoxylin and eosin stained cross sections. Reverse transcriptase PCR and immunohistochemistry were also performed to study the gene and protein level expression of angiogenic molecules. RESULTS: HRL has the ability to induce new vessel formation at the treated area and growth of the newly formed vessels and cellular morphological changes occur in a dose dependent manner. Increase in the tissue thickness at the treated area is suggestive of initiation of new capillary like structures. Elevated mRNA and protein level expression of VEGF165 and MMP2 along with the activation of ECs as demonstrated by the presence of CD34 expression supports the neovascularization potential of HRL. CONCLUSION: Angiogenic potential of HRL depends on the concentration and time of incubation and is involved in the activation of ECs along with the major interaction of VEGF 165 and MMP2. It is also observed that 3 μg of HRL exhibits maximum angiogenic potential at 72 h of incubation. Thus our data suggest that dose dependent angiogenic potential HRL could provide a novel role in angiogenic dependent therapeutics such as ischemia and wound healing conditions.info:eu-repo/semantics/openAccessSociedad de Biología de ChileBiological Research v.48 20152015-01-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602015000100029en10.1186/S40659-015-0021-Z
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	Angiogenesis HRL Human Recombinant Leptin CAM chorioallantoic membrane ECs endothelial cells VEGF vascular endothelial growth factor MMP matrix metalloproteinase Blood vessels CD34
spellingShingle	Angiogenesis HRL Human Recombinant Leptin CAM chorioallantoic membrane ECs endothelial cells VEGF vascular endothelial growth factor MMP matrix metalloproteinase Blood vessels CD34 Manjunathan,Reji Ragunathan,Malathi In ovo administration of human recombinant leptin shows dose dependent angiogenic effect on chicken chorioallantoic membrane
description	BACKGROUND: Leptin, the cytokine produced by white adipose tissue is known to regulate food energy homeostasis through its hypothalamic receptor. In vitro studies have demonstrated that leptin plays a major role in angiogenesis through binding to the receptor Ob-R present on ECs by stimulating and initiating new capillary like structures from ECs. Various in vivo studies indicate that leptin has diverse effect on angiogenesis. A few reports have showed that leptin exerts pro angiogenic effects while some suggested that it has antiangiogenic potential. It is theoretically highly important to understand the effect of leptin on angiogenesis to use as a therapeutic molecule in various angiogenesis related pathological conditions. Chicken chorio allantoic membrane (CAM) on 9th day of incubation was incubated with 1, 3 and 5 μg concentration of HRL for 72 h using gelatin sponge. Images where taken after every 24 h of incubation and analysed with Angioguant software. The treated area was observed under microscope and histological evaluation was performed for the same. Tissue thickness was calculated morphometrically from haematoxylin and eosin stained cross sections. Reverse transcriptase PCR and immunohistochemistry were also performed to study the gene and protein level expression of angiogenic molecules. RESULTS: HRL has the ability to induce new vessel formation at the treated area and growth of the newly formed vessels and cellular morphological changes occur in a dose dependent manner. Increase in the tissue thickness at the treated area is suggestive of initiation of new capillary like structures. Elevated mRNA and protein level expression of VEGF165 and MMP2 along with the activation of ECs as demonstrated by the presence of CD34 expression supports the neovascularization potential of HRL. CONCLUSION: Angiogenic potential of HRL depends on the concentration and time of incubation and is involved in the activation of ECs along with the major interaction of VEGF 165 and MMP2. It is also observed that 3 μg of HRL exhibits maximum angiogenic potential at 72 h of incubation. Thus our data suggest that dose dependent angiogenic potential HRL could provide a novel role in angiogenic dependent therapeutics such as ischemia and wound healing conditions.
author	Manjunathan,Reji Ragunathan,Malathi
author_facet	Manjunathan,Reji Ragunathan,Malathi
author_sort	Manjunathan,Reji
title	In ovo administration of human recombinant leptin shows dose dependent angiogenic effect on chicken chorioallantoic membrane
title_short	In ovo administration of human recombinant leptin shows dose dependent angiogenic effect on chicken chorioallantoic membrane
title_full	In ovo administration of human recombinant leptin shows dose dependent angiogenic effect on chicken chorioallantoic membrane
title_fullStr	In ovo administration of human recombinant leptin shows dose dependent angiogenic effect on chicken chorioallantoic membrane
title_full_unstemmed	In ovo administration of human recombinant leptin shows dose dependent angiogenic effect on chicken chorioallantoic membrane
title_sort	in ovo administration of human recombinant leptin shows dose dependent angiogenic effect on chicken chorioallantoic membrane
publisher	Sociedad de Biología de Chile
publishDate	2015
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602015000100029
work_keys_str_mv	AT manjunathanreji inovoadministrationofhumanrecombinantleptinshowsdosedependentangiogeniceffectonchickenchorioallantoicmembrane AT ragunathanmalathi inovoadministrationofhumanrecombinantleptinshowsdosedependentangiogeniceffectonchickenchorioallantoicmembrane
_version_	1718441534584520704

In ovo administration of human recombinant leptin shows dose dependent angiogenic effect on chicken chorioallantoic membrane

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