Skip to main content
Top
Published in: BMC Complementary Medicine and Therapies 1/2021

Open Access 01-12-2021 | Research article

Porcine placenta extract improves high-glucose-induced angiogenesis impairment

Authors: Chatchai Nensat, Worawat Songjang, Rutaiwan Tohtong, Tuangporn Suthiphongchai, Suchada Phimsen, Panthip Rattanasinganchan, Pornphimon Metheenukul, Sarawut Kumphune, Arunya Jiraviriyakul

Published in: BMC Complementary Medicine and Therapies | Issue 1/2021

Login to get access

Abstract

Background

High glucose (HG)-induced reactive oxygen species (ROS) overproduction impairs angiogenesis that is one pivotal factor of wound healing process. Angiogenesis impairment induces delayed wound healing, whereby it eventually leads to amputation in cases of poorly controlled diabetes with diabetic ulceration. Porcine placenta extract (PPE) is a natural waste product that comprises plenty of bioactive agents including growth factors and antioxidants. It was reported as an effective compound that prevents ROS generation. The goal of this study was to investigate the in vitro effect of PPE on HG-induced ROS-mediated angiogenesis impairment.

Methods

Primary endothelial cells (HUVECs) and endothelial cell line (EA.hy926) were treated with HG in the presence of PPE. The endothelial cells (ECs) viability, intracellular ROS generation, migration, and angiogenesis were determined by MTT assay, DCFDA reagent, wound healing assay, and tube formation assay, respectively. Additionally, the molecular mechanism of PPE on HG-induced angiogenesis impairment was investigated by Western blot. The angiogenic growth factor secretion was also investigated by the sandwich ELISA technique.

Results

HG in the presence of PPE significantly decreased intracellular ROS overproduction compared to HG alone. HG in the presence of PPE significantly increased ECs viability, migration, and angiogenesis compared to HG alone by showing recovery of PI3K/Akt/ERK1/2 activation. HG in the presence of PPE also decreased ECs apoptosis compared to HG alone by decreasing p53/Bax/cleaved caspase 9/cleaved caspase 3 levels and increasing Bcl 2 level.

Conclusion

PPE attenuated HG-induced intracellular ROS overproduction that improved ECs viability, proliferation, migration, and angiogenesis by showing recovery of PI3K/Akt/ERK1/2 activation and inhibition of ECs apoptosis. This study suggests PPE ameliorated HG-induced ROS-mediated angiogenesis impairment, whereby it potentially provides an alternative treatment for diabetic wounds.
Appendix
Available only for authorised users
Literature
1.
go back to reference Tandara AA, Mustoe TA. Oxygen in wound healing—more than a nutrient. World J Surg. 2004;28(3):294–300.CrossRef Tandara AA, Mustoe TA. Oxygen in wound healing—more than a nutrient. World J Surg. 2004;28(3):294–300.CrossRef
2.
go back to reference Li J, Zhang Y-P, Kirsner RS. Angiogenesis in wound repair: Angiogenic growth factors and the extracellular matrix. Microsc Res Tech. 2003;60(1):107–14.CrossRef Li J, Zhang Y-P, Kirsner RS. Angiogenesis in wound repair: Angiogenic growth factors and the extracellular matrix. Microsc Res Tech. 2003;60(1):107–14.CrossRef
3.
go back to reference Bauer SM, Bauer RJ, Velazquez OC. Angiogenesis, Vasculogenesis, and induction of healing in chronic wounds. Vasc Endovasc Surg. 2005;39(4):293–306.CrossRef Bauer SM, Bauer RJ, Velazquez OC. Angiogenesis, Vasculogenesis, and induction of healing in chronic wounds. Vasc Endovasc Surg. 2005;39(4):293–306.CrossRef
4.
go back to reference Tahergorabi Z, Khazaei M. Imbalance of angiogenesis in diabetic complications: the mechanisms. Int J Prev Med. 2012;3(12):827–38.CrossRef Tahergorabi Z, Khazaei M. Imbalance of angiogenesis in diabetic complications: the mechanisms. Int J Prev Med. 2012;3(12):827–38.CrossRef
5.
go back to reference Simo R, Carrasco E, Garcia-Ramirez M, Hernandez C. Angiogenic and Antiangiogenic factors in proliferative diabetic retinopathy. Curr Diab Rev. 2006;2(1):71–98.CrossRef Simo R, Carrasco E, Garcia-Ramirez M, Hernandez C. Angiogenic and Antiangiogenic factors in proliferative diabetic retinopathy. Curr Diab Rev. 2006;2(1):71–98.CrossRef
6.
go back to reference Polverini PJ. Angiogenesis and wound healing: basic discoveries, clinical implications, and therapeutic opportunities. Endod Top. 2011;24(1):130–45.CrossRef Polverini PJ. Angiogenesis and wound healing: basic discoveries, clinical implications, and therapeutic opportunities. Endod Top. 2011;24(1):130–45.CrossRef
7.
go back to reference Liu H, Yu S, Zhang H, Xu J. Angiogenesis impairment in diabetes: role of Methylglyoxal-induced receptor for advanced Glycation Endproducts, autophagy and vascular endothelial growth factor receptor 2. PLoS One. 2012;7(10):e46720.CrossRef Liu H, Yu S, Zhang H, Xu J. Angiogenesis impairment in diabetes: role of Methylglyoxal-induced receptor for advanced Glycation Endproducts, autophagy and vascular endothelial growth factor receptor 2. PLoS One. 2012;7(10):e46720.CrossRef
8.
go back to reference Crawford TN, Alfaro Iii DV, Kerrison JB, Jablon EP. Diabetic retinopathy and angiogenesis. Curr Diabetes Rev. 2009;5(1):8–13.CrossRef Crawford TN, Alfaro Iii DV, Kerrison JB, Jablon EP. Diabetic retinopathy and angiogenesis. Curr Diabetes Rev. 2009;5(1):8–13.CrossRef
9.
go back to reference Schieber M, Chandel NS. ROS function in redox signaling and oxidative stress. Curr Biol. 2014;24(10):R453–62.CrossRef Schieber M, Chandel NS. ROS function in redox signaling and oxidative stress. Curr Biol. 2014;24(10):R453–62.CrossRef
10.
go back to reference Fadini GP, Sartore S, Baesso I, Lenzi M, Agostini C, Tiengo A, et al. Endothelial progenitor cells and the diabetic paradox. Diabetes Care. 2006;29(3):714.CrossRef Fadini GP, Sartore S, Baesso I, Lenzi M, Agostini C, Tiengo A, et al. Endothelial progenitor cells and the diabetic paradox. Diabetes Care. 2006;29(3):714.CrossRef
11.
go back to reference Zhao Y, Singh RP. The role of anti-vascular endothelial growth factor (anti-VEGF) in the management of proliferative diabetic retinopathy. Drugs Context. 2018;7:–212532. Zhao Y, Singh RP. The role of anti-vascular endothelial growth factor (anti-VEGF) in the management of proliferative diabetic retinopathy. Drugs Context. 2018;7:–212532.
12.
go back to reference Abdallah W, Fawzi AA. Anti-VEGF therapy in proliferative diabetic retinopathy. Int Ophthalmol Clin. 2009;49(2):95–107.CrossRef Abdallah W, Fawzi AA. Anti-VEGF therapy in proliferative diabetic retinopathy. Int Ophthalmol Clin. 2009;49(2):95–107.CrossRef
13.
go back to reference Wu C-H, Chang G-Y, Chang W-C, Hsu C-T, Chen R-S. Wound healing effects of porcine placental extracts on rats with thermal injury. Br J Dermatol. 2003;148(2):236–45.CrossRef Wu C-H, Chang G-Y, Chang W-C, Hsu C-T, Chen R-S. Wound healing effects of porcine placental extracts on rats with thermal injury. Br J Dermatol. 2003;148(2):236–45.CrossRef
14.
go back to reference Heo JH, Heo Y, Lee HJ, Kim M, Shin HY. Topical anti-inflammatory and anti-oxidative effects of porcine placenta extracts on 2,4-dinitrochlorobenzene-induced contact dermatitis. BMC Complement Altern Med. 2018;18(1):331.CrossRef Heo JH, Heo Y, Lee HJ, Kim M, Shin HY. Topical anti-inflammatory and anti-oxidative effects of porcine placenta extracts on 2,4-dinitrochlorobenzene-induced contact dermatitis. BMC Complement Altern Med. 2018;18(1):331.CrossRef
15.
go back to reference Kongpol K, Nernpermpisooth N, Prompunt E, Kumphune S. Endothelial-cell-derived human secretory leukocyte protease inhibitor (SLPI) protects Cardiomyocytes against ischemia/reperfusion injury. Biomolecules. 2019;9(11):678.CrossRef Kongpol K, Nernpermpisooth N, Prompunt E, Kumphune S. Endothelial-cell-derived human secretory leukocyte protease inhibitor (SLPI) protects Cardiomyocytes against ischemia/reperfusion injury. Biomolecules. 2019;9(11):678.CrossRef
16.
go back to reference Shenouda SM, Widlansky ME, Chen K, Xu G, Holbrook M, Tabit CE, et al. Altered mitochondrial dynamics contributes to endothelial dysfunction in diabetes mellitus. Circulation. 2011;124(4):444–53.CrossRef Shenouda SM, Widlansky ME, Chen K, Xu G, Holbrook M, Tabit CE, et al. Altered mitochondrial dynamics contributes to endothelial dysfunction in diabetes mellitus. Circulation. 2011;124(4):444–53.CrossRef
17.
go back to reference Szewczyk A, Jarmuszkiewicz W, Koziel A, Sobieraj I, Nobik W, Lukasiak A, et al. Mitochondrial mechanisms of endothelial dysfunction. Pharmacol Rep. 2015;67(4):704–10.CrossRef Szewczyk A, Jarmuszkiewicz W, Koziel A, Sobieraj I, Nobik W, Lukasiak A, et al. Mitochondrial mechanisms of endothelial dysfunction. Pharmacol Rep. 2015;67(4):704–10.CrossRef
18.
go back to reference Zhu ZX, Cai WH, Wang T, Ye HB, Zhu YT, Chi LS, et al. bFGF-regulating MAPKs are involved in high glucose-mediated ROS production and delay of vascular endothelial cell migration. PLoS One. 2015;10(12):e0144495.CrossRef Zhu ZX, Cai WH, Wang T, Ye HB, Zhu YT, Chi LS, et al. bFGF-regulating MAPKs are involved in high glucose-mediated ROS production and delay of vascular endothelial cell migration. PLoS One. 2015;10(12):e0144495.CrossRef
19.
go back to reference Rezabakhsh A, Ahmadi M, Khaksar M, Montaseri A, Malekinejad H, Rahbarghazi R, et al. Rapamycin inhibits oxidative/nitrosative stress and enhances angiogenesis in high glucose-treated human umbilical vein endothelial cells: role of autophagy. Biomed Pharmacother. 2017;93:885–94.CrossRef Rezabakhsh A, Ahmadi M, Khaksar M, Montaseri A, Malekinejad H, Rahbarghazi R, et al. Rapamycin inhibits oxidative/nitrosative stress and enhances angiogenesis in high glucose-treated human umbilical vein endothelial cells: role of autophagy. Biomed Pharmacother. 2017;93:885–94.CrossRef
20.
go back to reference Zhang Z, Yang Z, Zhu B, Hu J, Liew CW, Zhang Y, et al. Increasing glucose 6-phosphate dehydrogenase activity restores redox balance in vascular endothelial cells exposed to high glucose. PLoS One. 2012;7(11):e49128.CrossRef Zhang Z, Yang Z, Zhu B, Hu J, Liew CW, Zhang Y, et al. Increasing glucose 6-phosphate dehydrogenase activity restores redox balance in vascular endothelial cells exposed to high glucose. PLoS One. 2012;7(11):e49128.CrossRef
21.
go back to reference Montezano AC, Touyz RM. Reactive oxygen species and endothelial function – role of nitric oxide synthase uncoupling and Nox Family Nicotinamide adenine dinucleotide phosphate oxidases. Basic Clin Pharmacol Toxicol. 2012;110(1):87–94.CrossRef Montezano AC, Touyz RM. Reactive oxygen species and endothelial function – role of nitric oxide synthase uncoupling and Nox Family Nicotinamide adenine dinucleotide phosphate oxidases. Basic Clin Pharmacol Toxicol. 2012;110(1):87–94.CrossRef
22.
go back to reference Gao L, Mann GE. Vascular NAD(P) H oxidase activation in diabetes: a double-edged sword in redox signalling. Cardiovasc Res. 2009;82(1):9–20.CrossRef Gao L, Mann GE. Vascular NAD(P) H oxidase activation in diabetes: a double-edged sword in redox signalling. Cardiovasc Res. 2009;82(1):9–20.CrossRef
23.
go back to reference Ding H, Aljofan M, Triggle CR. Oxidative stress and increased eNOS and NADPH oxidase expression in mouse microvessel endothelial cells. J Cell Physiol. 2007;212(3):682–9.CrossRef Ding H, Aljofan M, Triggle CR. Oxidative stress and increased eNOS and NADPH oxidase expression in mouse microvessel endothelial cells. J Cell Physiol. 2007;212(3):682–9.CrossRef
24.
go back to reference Du X, Matsumura T, Edelstein D, Rossetti L, Zsengellér Z, Szabó C, et al. Inhibition of GAPDH activity by poly (ADP-ribose) polymerase activates three major pathways of hyperglycemic damage in endothelial cells. J Clin Invest. 2003;112(7):1049–57.CrossRef Du X, Matsumura T, Edelstein D, Rossetti L, Zsengellér Z, Szabó C, et al. Inhibition of GAPDH activity by poly (ADP-ribose) polymerase activates three major pathways of hyperglycemic damage in endothelial cells. J Clin Invest. 2003;112(7):1049–57.CrossRef
25.
go back to reference Ighodaro OM. Molecular pathways associated with oxidative stress in diabetes mellitus. Biomed Pharmacother. 2018;108:656–62.CrossRef Ighodaro OM. Molecular pathways associated with oxidative stress in diabetes mellitus. Biomed Pharmacother. 2018;108:656–62.CrossRef
26.
go back to reference Pricci F, Leto G, Amadio L, Iacobini C, Cordone S, Catalano S, et al. Oxidative stress in diabetes-induced endothelial dysfunction involvement of nitric oxide and protein kinase C. Free Radic Biol Med. 2003;35(6):683–94.CrossRef Pricci F, Leto G, Amadio L, Iacobini C, Cordone S, Catalano S, et al. Oxidative stress in diabetes-induced endothelial dysfunction involvement of nitric oxide and protein kinase C. Free Radic Biol Med. 2003;35(6):683–94.CrossRef
27.
go back to reference Incalza MA, D'Oria R, Natalicchio A, Perrini S, Laviola L, Giorgino F. Oxidative stress and reactive oxygen species in endothelial dysfunction associated with cardiovascular and metabolic diseases. Vasc Pharmacol. 2018;100:1–19.CrossRef Incalza MA, D'Oria R, Natalicchio A, Perrini S, Laviola L, Giorgino F. Oxidative stress and reactive oxygen species in endothelial dysfunction associated with cardiovascular and metabolic diseases. Vasc Pharmacol. 2018;100:1–19.CrossRef
28.
go back to reference Li Q, Lin Y, Wang S, Zhang L, Guo L. GLP-1 inhibits high-glucose-induced oxidative injury of vascular endothelial cells. Sci Rep. 2017;7(1):8008.CrossRef Li Q, Lin Y, Wang S, Zhang L, Guo L. GLP-1 inhibits high-glucose-induced oxidative injury of vascular endothelial cells. Sci Rep. 2017;7(1):8008.CrossRef
29.
go back to reference Duan M-X, Zhou H, Wu Q-Q, Liu C, Xiao Y, Deng W, et al. Andrographolide protects against HG-induced inflammation, apoptosis, migration, and impairment of angiogenesis via PI3K/AKT-eNOS Signalling in HUVECs. Mediat Inflamm. 2019:6168340. Duan M-X, Zhou H, Wu Q-Q, Liu C, Xiao Y, Deng W, et al. Andrographolide protects against HG-induced inflammation, apoptosis, migration, and impairment of angiogenesis via PI3K/AKT-eNOS Signalling in HUVECs. Mediat Inflamm. 2019:6168340.
30.
go back to reference Zhou D-Y, Su Y, Gao P, Yang Q-H, Wang Z, Xu Q. Resveratrol ameliorates high glucose-induced oxidative stress injury in human umbilical vein endothelial cells by activating AMPK. Life Sci. 2015;136:94–9.CrossRef Zhou D-Y, Su Y, Gao P, Yang Q-H, Wang Z, Xu Q. Resveratrol ameliorates high glucose-induced oxidative stress injury in human umbilical vein endothelial cells by activating AMPK. Life Sci. 2015;136:94–9.CrossRef
31.
go back to reference Rezabakhsh A, Montazersaheb S, Nabat E, Hassanpour M, Montaseri A, Malekinejad H, et al. Effect of hydroxychloroquine on oxidative/nitrosative status and angiogenesis in endothelial cells under high glucose condition. BioImpacts. 2017;7(4):219–26.CrossRef Rezabakhsh A, Montazersaheb S, Nabat E, Hassanpour M, Montaseri A, Malekinejad H, et al. Effect of hydroxychloroquine on oxidative/nitrosative status and angiogenesis in endothelial cells under high glucose condition. BioImpacts. 2017;7(4):219–26.CrossRef
32.
go back to reference Han J, Mandal AK, Hiebert LM. Endothelial cell injury by high glucose and heparanase is prevented by insulin, heparin and basic fibroblast growth factor. Cardiovasc Diabetol. 2005;4(1):12.CrossRef Han J, Mandal AK, Hiebert LM. Endothelial cell injury by high glucose and heparanase is prevented by insulin, heparin and basic fibroblast growth factor. Cardiovasc Diabetol. 2005;4(1):12.CrossRef
33.
go back to reference Yang Z, Mo X, Gong Q, Pan Q, Yang X, Cai W, et al. Critical effect of VEGF in the process of endothelial cell apoptosis induced by high glucose. Apoptosis. 2008;13(11):1331–43.CrossRef Yang Z, Mo X, Gong Q, Pan Q, Yang X, Cai W, et al. Critical effect of VEGF in the process of endothelial cell apoptosis induced by high glucose. Apoptosis. 2008;13(11):1331–43.CrossRef
34.
go back to reference Song H, Wu F, Zhang Y, Zhang Y, Wang F, Jiang M, et al. Irisin promotes human umbilical vein endothelial cell proliferation through the ERK signaling pathway and partly suppresses high glucose-induced apoptosis. PLoS One. 2014;9(10):e110273.CrossRef Song H, Wu F, Zhang Y, Zhang Y, Wang F, Jiang M, et al. Irisin promotes human umbilical vein endothelial cell proliferation through the ERK signaling pathway and partly suppresses high glucose-induced apoptosis. PLoS One. 2014;9(10):e110273.CrossRef
35.
go back to reference Padhomchai Pumbthongthae PH, Sittiruk Roytrakul, Tavan Janvilisri, Puey Ounjai, editor Analysis of Protein Profile of Crude porcine placenta extract. The 50th National Graduate Research Conference; 7 2020; King Mongkut's Institute of Technology Ladkrabang, Bangkok. Padhomchai Pumbthongthae PH, Sittiruk Roytrakul, Tavan Janvilisri, Puey Ounjai, editor Analysis of Protein Profile of Crude porcine placenta extract. The 50th National Graduate Research Conference; 7 2020; King Mongkut's Institute of Technology Ladkrabang, Bangkok.
36.
go back to reference Xing Y, Lai J, Liu X, Zhang N, Ming J, Liu H, et al. Netrin-1 restores cell injury and impaired angiogenesis in vascular endothelial cells upon high glucose by PI3K/AKT-eNOS. J Mol Endocrinol. 2017;58(4):167.CrossRef Xing Y, Lai J, Liu X, Zhang N, Ming J, Liu H, et al. Netrin-1 restores cell injury and impaired angiogenesis in vascular endothelial cells upon high glucose by PI3K/AKT-eNOS. J Mol Endocrinol. 2017;58(4):167.CrossRef
37.
go back to reference Maamoun H, Benameur T, Pintus G, Munusamy S, Agouni A. Crosstalk Between Oxidative Stress and Endoplasmic Reticulum (ER) Stress in Endothelial Dysfunction and Aberrant Angiogenesis Associated With Diabetes: A Focus on the Protective Roles of Heme Oxygenase (HO)-1. Front Physiol. 2019;10:70.CrossRef Maamoun H, Benameur T, Pintus G, Munusamy S, Agouni A. Crosstalk Between Oxidative Stress and Endoplasmic Reticulum (ER) Stress in Endothelial Dysfunction and Aberrant Angiogenesis Associated With Diabetes: A Focus on the Protective Roles of Heme Oxygenase (HO)-1. Front Physiol. 2019;10:70.CrossRef
38.
go back to reference De Nigris V, Pujadas G, La Sala L, Testa R, Genovese S, Ceriello A. Short-term high glucose exposure impairs insulin signaling in endothelial cells. Cardiovasc Diabetol. 2015;14(1):114.CrossRef De Nigris V, Pujadas G, La Sala L, Testa R, Genovese S, Ceriello A. Short-term high glucose exposure impairs insulin signaling in endothelial cells. Cardiovasc Diabetol. 2015;14(1):114.CrossRef
39.
go back to reference Lamalice L, Boeuf FL, Huot J. Endothelial cell migration during angiogenesis. Circ Res. 2007;100(6):782–94.CrossRef Lamalice L, Boeuf FL, Huot J. Endothelial cell migration during angiogenesis. Circ Res. 2007;100(6):782–94.CrossRef
40.
go back to reference Wu N, Shen H, Liu H, Wang Y, Bai Y, Han P. Acute blood glucose fluctuation enhances rat aorta endothelial cell apoptosis, oxidative stress and pro-inflammatory cytokine expression in vivo. Cardiovasc Diabetol. 2016;15(1):109.CrossRef Wu N, Shen H, Liu H, Wang Y, Bai Y, Han P. Acute blood glucose fluctuation enhances rat aorta endothelial cell apoptosis, oxidative stress and pro-inflammatory cytokine expression in vivo. Cardiovasc Diabetol. 2016;15(1):109.CrossRef
41.
go back to reference Hou Q, Lei M, Hu K, Wang M. The effects of high glucose levels on reactive oxygen species-induced apoptosis and involved signaling in human vascular endothelial cells. Cardiovasc Toxicol. 2015;15(2):140–6.CrossRef Hou Q, Lei M, Hu K, Wang M. The effects of high glucose levels on reactive oxygen species-induced apoptosis and involved signaling in human vascular endothelial cells. Cardiovasc Toxicol. 2015;15(2):140–6.CrossRef
42.
go back to reference Ucuzian AA, Gassman AA, East AT, Greisler HP. Molecular mediators of angiogenesis. J Burn Care Res. 2010;31(1):158–75.CrossRef Ucuzian AA, Gassman AA, East AT, Greisler HP. Molecular mediators of angiogenesis. J Burn Care Res. 2010;31(1):158–75.CrossRef
43.
go back to reference Xie D, Ju D, Speyer C, Gorski D, Kosir MA. Strategic endothelial cell tube formation assay: comparing extracellular matrix and growth factor reduced extracellular matrix. J Vis Exp. 2016;114:54074. Xie D, Ju D, Speyer C, Gorski D, Kosir MA. Strategic endothelial cell tube formation assay: comparing extracellular matrix and growth factor reduced extracellular matrix. J Vis Exp. 2016;114:54074.
Metadata
Title
Porcine placenta extract improves high-glucose-induced angiogenesis impairment
Authors
Chatchai Nensat
Worawat Songjang
Rutaiwan Tohtong
Tuangporn Suthiphongchai
Suchada Phimsen
Panthip Rattanasinganchan
Pornphimon Metheenukul
Sarawut Kumphune
Arunya Jiraviriyakul
Publication date
01-12-2021
Publisher
BioMed Central
Published in
BMC Complementary Medicine and Therapies / Issue 1/2021
Electronic ISSN: 2662-7671
DOI
https://doi.org/10.1186/s12906-021-03243-z

Other articles of this Issue 1/2021

BMC Complementary Medicine and Therapies 1/2021 Go to the issue