Abstract
Most evidence indicates that nitric oxide plays a role in normal wound repair; however, involvement of inducible nitric oxide synthase (iNOS) has not been established. Experiments were carried out to determine the requirement for iNOS in closing excisional wounds. Wound closure was delayed by 31% in iNOS knockout mice compared with wild-type animals. An identical delay in wound closure was observed in wild-type mice given a continuous infusion of the partially selective iNOS inhibitor N6-(iminoethyl)-L-lysine. Delayed wound healing in iNOS-deficient mice was completely reversed by a single application of an adenoviral vector containing human iNOS cDNA (AdiNOS) at the time of wounding. Reverse transcription PCR identified iNOS mRNA expression in wild-type mice peaking 4-6 d after wounding, and confirmed expression of human iNOS in the adenoviral vector containing human iNOS cDNA-treated animals. These results establish the key role of iNOS in wound closure, and suggest a gene therapy strategy to improve wound healing in iNOS-deficient states such as diabetes, and during steroid treatment.
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Selected References
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- Albina J. E., Mills C. D., Henry W. L., Jr, Caldwell M. D. Temporal expression of different pathways of 1-arginine metabolism in healing wounds. J Immunol. 1990 May 15;144(10):3877–3880. [PubMed] [Google Scholar]
- Barbul A., Lazarou S. A., Efron D. T., Wasserkrug H. L., Efron G. Arginine enhances wound healing and lymphocyte immune responses in humans. Surgery. 1990 Aug;108(2):331–337. [PubMed] [Google Scholar]
- Becker W. K., Shippee R. L., McManus A. T., Mason A. D., Jr, Pruitt B. A., Jr Kinetics of nitrogen oxide production following experimental thermal injury in rats. J Trauma. 1993 Jun;34(6):855–862. doi: 10.1097/00005373-199306000-00017. [DOI] [PubMed] [Google Scholar]
- Benrath J., Zimmermann M., Gillardon F. Substance P and nitric oxide mediate would healing of ultraviolet photodamaged rat skin: evidence for an effect of nitric oxide on keratinocyte proliferation. Neurosci Lett. 1995 Nov 10;200(1):17–20. doi: 10.1016/0304-3940(95)12062-9. [DOI] [PubMed] [Google Scholar]
- Carter E. A., Derojas-Walker T., Tamir S., Tannenbaum S. R., Yu Y. M., Tompkins R. G. Nitric oxide production is intensely and persistently increased in tissue by thermal injury. Biochem J. 1994 Nov 15;304(Pt 1):201–204. doi: 10.1042/bj3040201. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chinnadurai G., Chinnadurai S., Brusca J. Physical mapping of a large-plaque mutation of adenovirus type 2. J Virol. 1979 Nov;32(2):623–628. doi: 10.1128/jvi.32.2.623-628.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Connor J. R., Manning P. T., Settle S. L., Moore W. M., Jerome G. M., Webber R. K., Tjoeng F. S., Currie M. G. Suppression of adjuvant-induced arthritis by selective inhibition of inducible nitric oxide synthase. Eur J Pharmacol. 1995 Jan 24;273(1-2):15–24. doi: 10.1016/0014-2999(94)00672-t. [DOI] [PubMed] [Google Scholar]
- Geller D. A., Lowenstein C. J., Shapiro R. A., Nussler A. K., Di Silvio M., Wang S. C., Nakayama D. K., Simmons R. L., Snyder S. H., Billiar T. R. Molecular cloning and expression of inducible nitric oxide synthase from human hepatocytes. Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3491–3495. doi: 10.1073/pnas.90.8.3491. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Geller D. A., Nussler A. K., Di Silvio M., Lowenstein C. J., Shapiro R. A., Wang S. C., Simmons R. L., Billiar T. R. Cytokines, endotoxin, and glucocorticoids regulate the expression of inducible nitric oxide synthase in hepatocytes. Proc Natl Acad Sci U S A. 1993 Jan 15;90(2):522–526. doi: 10.1073/pnas.90.2.522. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Graham F. L., Smiley J., Russell W. C., Nairn R. Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol. 1977 Jul;36(1):59–74. doi: 10.1099/0022-1317-36-1-59. [DOI] [PubMed] [Google Scholar]
- Heck D. E., Laskin D. L., Gardner C. R., Laskin J. D. Epidermal growth factor suppresses nitric oxide and hydrogen peroxide production by keratinocytes. Potential role for nitric oxide in the regulation of wound healing. J Biol Chem. 1992 Oct 25;267(30):21277–21280. [PubMed] [Google Scholar]
- Kirk S. J., Hurson M., Regan M. C., Holt D. R., Wasserkrug H. L., Barbul A. Arginine stimulates wound healing and immune function in elderly human beings. Surgery. 1993 Aug;114(2):155–160. [PubMed] [Google Scholar]
- Leibovich S. J., Polverini P. J., Fong T. W., Harlow L. A., Koch A. E. Production of angiogenic activity by human monocytes requires an L-arginine/nitric oxide-synthase-dependent effector mechanism. Proc Natl Acad Sci U S A. 1994 May 10;91(10):4190–4194. doi: 10.1073/pnas.91.10.4190. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MacMicking J. D., Nathan C., Hom G., Chartrain N., Fletcher D. S., Trumbauer M., Stevens K., Xie Q. W., Sokol K., Hutchinson N. Altered responses to bacterial infection and endotoxic shock in mice lacking inducible nitric oxide synthase. Cell. 1995 May 19;81(4):641–650. doi: 10.1016/0092-8674(95)90085-3. [DOI] [PubMed] [Google Scholar]
- Morbidelli L., Chang C. H., Douglas J. G., Granger H. J., Ledda F., Ziche M. Nitric oxide mediates mitogenic effect of VEGF on coronary venular endothelium. Am J Physiol. 1996 Jan;270(1 Pt 2):H411–H415. doi: 10.1152/ajpheart.1996.270.1.H411. [DOI] [PubMed] [Google Scholar]
- Morris S. M., Jr, Billiar T. R. New insights into the regulation of inducible nitric oxide synthesis. Am J Physiol. 1994 Jun;266(6 Pt 1):E829–E839. doi: 10.1152/ajpendo.1994.266.6.E829. [DOI] [PubMed] [Google Scholar]
- Noiri E., Peresleni T., Srivastava N., Weber P., Bahou W. F., Peunova N., Goligorsky M. S. Nitric oxide is necessary for a switch from stationary to locomoting phenotype in epithelial cells. Am J Physiol. 1996 Mar;270(3 Pt 1):C794–C802. doi: 10.1152/ajpcell.1996.270.3.C794. [DOI] [PubMed] [Google Scholar]
- Schaffer M. R., Tantry U., Gross S. S., Wasserburg H. L., Barbul A. Nitric oxide regulates wound healing. J Surg Res. 1996 Jun;63(1):237–240. doi: 10.1006/jsre.1996.0254. [DOI] [PubMed] [Google Scholar]
- Schäffer M. R., Tantry U., Efron P. A., Ahrendt G. M., Thornton F. J., Barbul A. Diabetes-impaired healing and reduced wound nitric oxide synthesis: a possible pathophysiologic correlation. Surgery. 1997 May;121(5):513–519. doi: 10.1016/s0039-6060(97)90105-7. [DOI] [PubMed] [Google Scholar]
- Shabani M., Pulfer S. K., Bulgrin J. P., Smith D. J. Enhancement of wound repair with a topically applied nitric oxide-releasing polymer. Wound Repair Regen. 1996 Jul–Sep;4(3):353–362. doi: 10.1046/j.1524-475X.1996.40312.x. [DOI] [PubMed] [Google Scholar]
- Tashiro M., Kawakami Y., Abe R., Han W., Hata D., Sugie K., Yao L., Kawakami T. Increased secretion of TNF-alpha by costimulation of mast cells via CD28 and Fc epsilon RI. J Immunol. 1997 Mar 1;158(5):2382–2389. [PubMed] [Google Scholar]
- Tzeng E., Billiar T. R., Robbins P. D., Loftus M., Stuehr D. J. Expression of human inducible nitric oxide synthase in a tetrahydrobiopterin (H4B)-deficient cell line: H4B promotes assembly of enzyme subunits into an active dimer. Proc Natl Acad Sci U S A. 1995 Dec 5;92(25):11771–11775. doi: 10.1073/pnas.92.25.11771. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tzeng E., Shears L. L., 2nd, Robbins P. D., Pitt B. R., Geller D. A., Watkins S. C., Simmons R. L., Billiar T. R. Vascular gene transfer of the human inducible nitric oxide synthase: characterization of activity and effects on myointimal hyperplasia. Mol Med. 1996 Mar;2(2):211–225. [PMC free article] [PubMed] [Google Scholar]
- Ziche M., Morbidelli L., Masini E., Amerini S., Granger H. J., Maggi C. A., Geppetti P., Ledda F. Nitric oxide mediates angiogenesis in vivo and endothelial cell growth and migration in vitro promoted by substance P. J Clin Invest. 1994 Nov;94(5):2036–2044. doi: 10.1172/JCI117557. [DOI] [PMC free article] [PubMed] [Google Scholar]