Synergistic effect of substance P with epidermal growth factor on epithelial migration in rabbit cornea

Masatsugu Nakamura, Teruo Nishida, Keiko Ofuji, Ted W. Reid, Mark J Mannis, Christopher J Murphy

Research output: Contribution to journalArticle

69 Citations (Scopus)

Abstract

In order to investigate the role of neural regulation in corneal epithelial healing, we examined the effect of substance P (SP) on corneal epithelial migration using an organ culture system of rabbit corneas. We investigated the synergistic effects of SP with (1) growth factors: epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), and transforming growth factor-β (TGF-β); (2) extracellular matrix proteins: fibronectin, vitronectin, laminin, and collagen type IV; and (3) cytokines: interleukin- 1α (IL-1α), IL-1β, and interleukin-6 (IL-6). Rabbit corneal blocks were cultured in the absence or presence of various reagents for 24 hr. The corneal blocks were then fixed, dehydrated, embedded in paraffin and stained by hematoxylin-eosin, and the length of the path of epithelial migration was measured. The addition of SP alone, at concentrations up to 50 μg ml-1, did not affect epithelial migration. EGF, fibronectin, vitronectin, collagen type IV, and IL-6 stimulated epithelial migration, but bFGF, TGF-β, laminin, IL-1α, and IL-1β did not. The stimulatory effect of EGF on the epithelial migration was enhanced by the presence of SP. This synergistic effect of SP and EGF on corneal epithelial migration was abolished by the addition of an SP antagonist or enkephalinase. Other neurotransmitters (vasoactive intestinal peptide, calcitonin gene-related peptide, acetylcholine chloride, norepinephrine, serotonin) and tachykinins (neurokinin A, neurokinin B, kassinin, eledoisin, physalaemin) were examined, but none exhibited a synergistic effect with EGF. Interestingly, EGF alone stimulated the incorporation of 3H-thymidine into corneal epithelial cells, but the addition of SP with EGF did not enhance this effect. These results demonstrate that SP enhanced the EGF stimulation of corneal epithelial migration in vitro in a specific manner, suggesting a possible role of SP as a modulator of epithelial wound healing.

Original languageEnglish (US)
Pages (from-to)321-329
Number of pages9
JournalExperimental Eye Research
Volume65
Issue number3
DOIs
StatePublished - Sep 1997

Fingerprint

Substance P
Epidermal Growth Factor
Cornea
Rabbits
Interleukin-1
Vitronectin
Transforming Growth Factors
Fibroblast Growth Factor 2
Fibronectins
Kassinin
Interleukin-6
Physalaemin
Eledoisin
Neurokinin B
Neurokinin A
Tachykinins
Neprilysin
Collagen Type III
Collagen Type IV
Calcitonin Gene-Related Peptide

Keywords

  • Cornea
  • Epidermal growth factor
  • Epithelial migration
  • Extracellular matrix proteins
  • Growth factors
  • Interleukins
  • Substance P

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems

Cite this

Synergistic effect of substance P with epidermal growth factor on epithelial migration in rabbit cornea. / Nakamura, Masatsugu; Nishida, Teruo; Ofuji, Keiko; Reid, Ted W.; Mannis, Mark J; Murphy, Christopher J.

In: Experimental Eye Research, Vol. 65, No. 3, 09.1997, p. 321-329.

Research output: Contribution to journalArticle

Nakamura, Masatsugu ; Nishida, Teruo ; Ofuji, Keiko ; Reid, Ted W. ; Mannis, Mark J ; Murphy, Christopher J. / Synergistic effect of substance P with epidermal growth factor on epithelial migration in rabbit cornea. In: Experimental Eye Research. 1997 ; Vol. 65, No. 3. pp. 321-329.
@article{cbc67fa304b741da9ffc2c19c00f7995,
title = "Synergistic effect of substance P with epidermal growth factor on epithelial migration in rabbit cornea",
abstract = "In order to investigate the role of neural regulation in corneal epithelial healing, we examined the effect of substance P (SP) on corneal epithelial migration using an organ culture system of rabbit corneas. We investigated the synergistic effects of SP with (1) growth factors: epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), and transforming growth factor-β (TGF-β); (2) extracellular matrix proteins: fibronectin, vitronectin, laminin, and collagen type IV; and (3) cytokines: interleukin- 1α (IL-1α), IL-1β, and interleukin-6 (IL-6). Rabbit corneal blocks were cultured in the absence or presence of various reagents for 24 hr. The corneal blocks were then fixed, dehydrated, embedded in paraffin and stained by hematoxylin-eosin, and the length of the path of epithelial migration was measured. The addition of SP alone, at concentrations up to 50 μg ml-1, did not affect epithelial migration. EGF, fibronectin, vitronectin, collagen type IV, and IL-6 stimulated epithelial migration, but bFGF, TGF-β, laminin, IL-1α, and IL-1β did not. The stimulatory effect of EGF on the epithelial migration was enhanced by the presence of SP. This synergistic effect of SP and EGF on corneal epithelial migration was abolished by the addition of an SP antagonist or enkephalinase. Other neurotransmitters (vasoactive intestinal peptide, calcitonin gene-related peptide, acetylcholine chloride, norepinephrine, serotonin) and tachykinins (neurokinin A, neurokinin B, kassinin, eledoisin, physalaemin) were examined, but none exhibited a synergistic effect with EGF. Interestingly, EGF alone stimulated the incorporation of 3H-thymidine into corneal epithelial cells, but the addition of SP with EGF did not enhance this effect. These results demonstrate that SP enhanced the EGF stimulation of corneal epithelial migration in vitro in a specific manner, suggesting a possible role of SP as a modulator of epithelial wound healing.",
keywords = "Cornea, Epidermal growth factor, Epithelial migration, Extracellular matrix proteins, Growth factors, Interleukins, Substance P",
author = "Masatsugu Nakamura and Teruo Nishida and Keiko Ofuji and Reid, {Ted W.} and Mannis, {Mark J} and Murphy, {Christopher J}",
year = "1997",
month = "9",
doi = "10.1006/exer.1997.0345",
language = "English (US)",
volume = "65",
pages = "321--329",
journal = "Experimental Eye Research",
issn = "0014-4835",
publisher = "Academic Press Inc.",
number = "3",

}

TY - JOUR

T1 - Synergistic effect of substance P with epidermal growth factor on epithelial migration in rabbit cornea

AU - Nakamura, Masatsugu

AU - Nishida, Teruo

AU - Ofuji, Keiko

AU - Reid, Ted W.

AU - Mannis, Mark J

AU - Murphy, Christopher J

PY - 1997/9

Y1 - 1997/9

N2 - In order to investigate the role of neural regulation in corneal epithelial healing, we examined the effect of substance P (SP) on corneal epithelial migration using an organ culture system of rabbit corneas. We investigated the synergistic effects of SP with (1) growth factors: epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), and transforming growth factor-β (TGF-β); (2) extracellular matrix proteins: fibronectin, vitronectin, laminin, and collagen type IV; and (3) cytokines: interleukin- 1α (IL-1α), IL-1β, and interleukin-6 (IL-6). Rabbit corneal blocks were cultured in the absence or presence of various reagents for 24 hr. The corneal blocks were then fixed, dehydrated, embedded in paraffin and stained by hematoxylin-eosin, and the length of the path of epithelial migration was measured. The addition of SP alone, at concentrations up to 50 μg ml-1, did not affect epithelial migration. EGF, fibronectin, vitronectin, collagen type IV, and IL-6 stimulated epithelial migration, but bFGF, TGF-β, laminin, IL-1α, and IL-1β did not. The stimulatory effect of EGF on the epithelial migration was enhanced by the presence of SP. This synergistic effect of SP and EGF on corneal epithelial migration was abolished by the addition of an SP antagonist or enkephalinase. Other neurotransmitters (vasoactive intestinal peptide, calcitonin gene-related peptide, acetylcholine chloride, norepinephrine, serotonin) and tachykinins (neurokinin A, neurokinin B, kassinin, eledoisin, physalaemin) were examined, but none exhibited a synergistic effect with EGF. Interestingly, EGF alone stimulated the incorporation of 3H-thymidine into corneal epithelial cells, but the addition of SP with EGF did not enhance this effect. These results demonstrate that SP enhanced the EGF stimulation of corneal epithelial migration in vitro in a specific manner, suggesting a possible role of SP as a modulator of epithelial wound healing.

AB - In order to investigate the role of neural regulation in corneal epithelial healing, we examined the effect of substance P (SP) on corneal epithelial migration using an organ culture system of rabbit corneas. We investigated the synergistic effects of SP with (1) growth factors: epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), and transforming growth factor-β (TGF-β); (2) extracellular matrix proteins: fibronectin, vitronectin, laminin, and collagen type IV; and (3) cytokines: interleukin- 1α (IL-1α), IL-1β, and interleukin-6 (IL-6). Rabbit corneal blocks were cultured in the absence or presence of various reagents for 24 hr. The corneal blocks were then fixed, dehydrated, embedded in paraffin and stained by hematoxylin-eosin, and the length of the path of epithelial migration was measured. The addition of SP alone, at concentrations up to 50 μg ml-1, did not affect epithelial migration. EGF, fibronectin, vitronectin, collagen type IV, and IL-6 stimulated epithelial migration, but bFGF, TGF-β, laminin, IL-1α, and IL-1β did not. The stimulatory effect of EGF on the epithelial migration was enhanced by the presence of SP. This synergistic effect of SP and EGF on corneal epithelial migration was abolished by the addition of an SP antagonist or enkephalinase. Other neurotransmitters (vasoactive intestinal peptide, calcitonin gene-related peptide, acetylcholine chloride, norepinephrine, serotonin) and tachykinins (neurokinin A, neurokinin B, kassinin, eledoisin, physalaemin) were examined, but none exhibited a synergistic effect with EGF. Interestingly, EGF alone stimulated the incorporation of 3H-thymidine into corneal epithelial cells, but the addition of SP with EGF did not enhance this effect. These results demonstrate that SP enhanced the EGF stimulation of corneal epithelial migration in vitro in a specific manner, suggesting a possible role of SP as a modulator of epithelial wound healing.

KW - Cornea

KW - Epidermal growth factor

KW - Epithelial migration

KW - Extracellular matrix proteins

KW - Growth factors

KW - Interleukins

KW - Substance P

UR - http://www.scopus.com/inward/record.url?scp=0030866765&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030866765&partnerID=8YFLogxK

U2 - 10.1006/exer.1997.0345

DO - 10.1006/exer.1997.0345

M3 - Article

VL - 65

SP - 321

EP - 329

JO - Experimental Eye Research

JF - Experimental Eye Research

SN - 0014-4835

IS - 3

ER -