Nitric oxide in the healing wound: A time-course study
Nitric oxide in the healing wound: A time-course study
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DOI：
10.1006/jsre.2001.6261
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发表时间：
2001-11-01
影响因子：
2.2
通讯作者：
Barbul, A
中科院分区：
文献类型：
--
作者：
Lee, RH;Efron, D;Barbul, A
导言。我们研究了创面愈合过程中一氧化氮表达的时程以及参与其合成的细胞群。对24只Lewis大鼠进行了聚乙烯醇海绵皮下植入实验。分别于伤后第1、3、5、7、10、14、35天处死大鼠。测定了在N-G-L-单甲基精氨酸(L-NMMA)作用下，海绵中H-3标记的精氨酸向H-3标记的瓜氨酸的转化。用Greiss法测定血浆和创面液中硝酸盐/亚硝酸盐(NOx)的含量。Northern分析和逆转录聚合酶链式反应(RT-PCR)检测诱导型一氧化氮合酶(INOS)基因表达。用双标记流式细胞术检测到特定的创面细胞群中存在诱导型一氧化氮合酶。一氧化氮合酶(NOS)活性在伤后24 h达高峰(每毫克海绵含37.7+/-0.9 mU/L瓜氨酸)，此后持续下降。对一氧化氮合酶活性的抑制率在第1~7天最高(70~80%)。到第10天，这一比例下降到50%，到第14-35天，下降到25%。INOS基因表达与一氧化氮合酶生化活性呈正相关。RT-PCR证实在伤后10天内有低水平的表达。创伤后血浆NOx水平始终维持在2 2.6+/-1.3~2 9.3+/-1.5um的窄幅范围内，而创面液中的NOx水平从伤后第1天的2 7+/-3.8升至伤后第14天的107.2+/-10.0。创伤后第1天和第3天的巨噬细胞可检测到诱导型一氧化氮合酶。我们的研究结果表明，在皮肤创伤愈合的早期，一氧化氮合酶活性最高，并在创伤后10天内持续产生。INOS基因表达与NOS生化活性呈正相关。血浆NOx保持不变，而创口液NOx稳步增加，在第14天达到高峰。创面巨噬细胞似乎是创面愈合早期一氧化氮的来源。(C)2001年学术出版社。
Introduction. We studied the time course of nitric oxide expression in the healing wound and the cell populations responsible for its synthesis.Methods. Twenty four Lewis rats underwent subcutaneous implantation of polyvinyl alcohol sponges. Rats were sacrificed in groups of three on days 1, 3, 5, 7, 10, 14, and 35 after wounding. The conversion of H-3-labeled arginine to H-3-labeled citrulline, with or without N-G-L-monomethyl-arginine (L-NMMA) in harvested sponges, was measured. Nitrate/nitrite (NOx) in plasma and wound fluid was quantified by Greiss reaction. Inducible nitric oxide synthase (iNOS) gene expression was determined by Northern analysis and reverse transcriptase-polymerase chain reaction (RT-PCR). Inducible NOS was identified in specific wound cell populations by dual-label flow cytometry.Results. Nitric oxide synthase (NOS) activity peaked at 24 h after wounding (37.7 +/- 0.9 mu mol citrulline per milligram sponge), with a steady decline thereafter. Percentage inhibition of NOS activity by L-NMMA was highest on days 1-7 (70-80%). This declined to 50% by day 10 and to 25% by days 14-35. The iNOS gene expression paralleled NOS biochemical activity. RT-PCR confirmed low-level expression up to 10 days after wounding. Plasma NOx levels remained within a narrow range of 22.6 +/- 1.3 to 29.3 +/- 1.5 muM throughout the postwounding period, while corresponding levels in wound fluid (muM) increased steadily from 27 +/- 3.8 on day 1 to 107.2 +/- 10.0 on day 14. Inducible NOS expression was detectable by fluorescence-activated cell sorting in wound macrophages on days 1 and 3 after wounding.Conclusions. Our findings suggest maximal NOS activity early in cutaneous wound healing, with sustained production up to 10 days after wounding. NOS biochemical activity was paralleled by iNOS gene expression. Plasma NOx remained constant, while wound fluid NOx increased steadily to peak at day 14. Wound macrophages appear to he a source of nitric oxide production in the early phase of wound healing. (C) 2001 Academic Press.