Shi H P, Most D, Efron D T, Tantry U, Fischel M H, Barbul A
Department of Surgery, Sinai Hospital of Baltimore, MD 21215, USA.
Surgery. 2001 Aug;130(2):225-9. doi: 10.1067/msy.2001.115837.
We have previously shown that the blockade of nitric oxide (NO) synthesis impairs wound healing, in particular collagen synthesis. Conversely, impaired wound healing is accompanied by decreased wound NO synthesis. Fibroblast collagen synthesis, proliferation, and fibroblast-mediated matrix contraction are critical to wound healing. We examined the wound healing-related phenotypic changes that are induced by the loss of inducible nitric oxide synthase (iNOS) gene function in fibroblasts.
Dermal fibroblasts were obtained from 8- to 12-week-old iNOS--knock out (KO; C57BL/Ai-[KO] Nos2 N5) and wild type mice by an explant technique and used after 1 to 3 passages. Proliferation ([(3)H]-thymidine incorporation) and collagen synthesis ([(3)H]-proline incorporation into collagenase-sensitive protein) were studied after stimulation with 10% fetal bovine serum. Matrix remodeling was assessed by the measurement of the contraction of fibroblast-populated collagen lattices.
iNOS-KO fibroblasts proliferated more slowly, synthesized less collagen, and contracted fibroblast-populated collagen lattices more slowly than wild-type fibroblast. Collagen synthesis was restored to normal in KO fibroblasts in response to NO donors (s-nitroso-N-acetylpenicillamine).
iNOS deficiency causes significant impairment in wound healing-related properties of fibroblasts, which suggests that NO plays an important role in wound healing.
我们之前已经表明,一氧化氮(NO)合成的阻断会损害伤口愈合,尤其是胶原蛋白的合成。相反,伤口愈合受损伴随着伤口NO合成的减少。成纤维细胞胶原蛋白合成、增殖以及成纤维细胞介导的基质收缩对伤口愈合至关重要。我们研究了成纤维细胞中诱导型一氧化氮合酶(iNOS)基因功能丧失所诱导的与伤口愈合相关的表型变化。
通过外植技术从8至12周龄的iNOS基因敲除(KO;C57BL/Ai-[KO] Nos2 N5)小鼠和野生型小鼠中获取真皮成纤维细胞,并在传代1至3次后使用。在用10%胎牛血清刺激后,研究增殖([³H] - 胸腺嘧啶掺入)和胶原蛋白合成([³H] - 脯氨酸掺入胶原酶敏感蛋白)。通过测量成纤维细胞填充的胶原晶格的收缩来评估基质重塑。
与野生型成纤维细胞相比,iNOS基因敲除的成纤维细胞增殖更慢,合成的胶原蛋白更少,并且使成纤维细胞填充的胶原晶格收缩更慢。给予NO供体(s - 亚硝基 - N - 乙酰青霉胺)后,基因敲除的成纤维细胞中的胶原蛋白合成恢复正常。
iNOS缺乏导致成纤维细胞与伤口愈合相关特性的显著损害,这表明NO在伤口愈合中起重要作用。