Lee R H, Efron D T, Tantry U, Stuelten C, Moldawer L L, Abouhamze A, Barbul A
Department of Surgery, Sinai Hospital of Baltimore, 2435 West Belvedere Avenue, Baltimore, MD 21215, USA.
Wound Repair Regen. 2000 Nov-Dec;8(6):547-53. doi: 10.1046/j.1524-475x.2000.00547.x.
Exogenous administration of tumor necrosis factor-alpha has been shown to both enhance and attenuate cutaneous healing in a dose-dependent manner. We examined the effects of tumor necrosis factor inhibition in the healing wound by both systemic and local administration of tumor necrosis factor-binding protein. Male Balb/C mice underwent dorsal skin incision with subcutaneous implantation of 20 mg polyvinyl alcohol sponges (4 per animal). In Experiment I, one group (n = 20) received intraperitoneal injections of tumor necrosis factor-binding protein (3 mg/kg) at the time of wounding, while another group (n = 20) received saline. Four animals from each group were euthanized on days 1, 3, 5, 7, and 14 postwounding. In Experiment II, one group (n = 10) received an intraperitoneal injection of tumor necrosis factor-binding protein (3 mg/kg) at the time of wounding and every third day thereafter. Another group (n = 10) received an intraperitoneal injection of saline at the time of wounding and every third day thereafter. In Experiment III, one group received a single intraperitoneal injection of tumor necrosis factor-binding protein (3 mg/kg) at the time of wounding (n = 7), or on postwounding day 4 (n = 7), or day 7 (n = 7). Another group received saline injections at the time of wounding (n = 7), or on postwounding days 4 or 7 (n = 7, respectively). All animals in Experiments II and III were killed at postwounding day 14. Wound breaking strengths were assessed using a tensiometer. Wound fluid collected from the implanted sponges was assayed for tumor necrosis factor-alpha and tumor necrosis factor-binding protein levels using a biological assay and enzyme-linked immunosorbent assay, respectively. Collagen gene expression in sponge granulomata was assessed by Northern analysis. Collagen deposition in sponges was quantified by measuring hydroxyproline content. Wounds were significantly weaker in the animals that received repeated injections of tumor necrosis factor-binding protein with a mean wound breaking strength of 93.1 g vs. 186.6 g in controls (p < 0.05). Wound breaking strength in groups that received a single injection of tumor necrosis factor-binding protein on either day 0, 4, or 7 postwounding were no different than their respective controls. There was no difference in the mean hydroxyproline content of sponges between any of the tumor necrosis factor-binding protein groups and their respective controls. Northern analysis for collagen I and III expression also revealed no differences. These data indicate that continued systemic administration of tumor necrosis factor-binding protein resulted in significantly weaker wounds with no corresponding differences in wound collagen content, and collagen gene expression. This suggests that tumor necrosis factor-alpha inhibition throughout healing leads to a qualitatively impaired wound without a quantitative alteration in collagen deposition.
已表明，外源性给予肿瘤坏死因子-α会以剂量依赖的方式增强和减弱皮肤愈合。我们通过全身和局部给予肿瘤坏死因子结合蛋白，研究了肿瘤坏死因子抑制对愈合伤口的影响。雄性Balb/C小鼠进行背部皮肤切口，并皮下植入20mg聚乙烯醇海绵（每只动物4块）。在实验I中，一组（n = 20）在受伤时腹腔注射肿瘤坏死因子结合蛋白（3mg/kg），而另一组（n = 20）注射生理盐水。每组4只动物在受伤后第1、3、5、7和14天安乐死。在实验II中，一组（n = 10）在受伤时及此后每三天腹腔注射肿瘤坏死因子结合蛋白（3mg/kg）。另一组（n = 10）在受伤时及此后每三天腹腔注射生理盐水。在实验III中，一组在受伤时（n = 7）、受伤后第4天（n = 7）或第7天（n = 7）单次腹腔注射肿瘤坏死因子结合蛋白（3mg/kg）。另一组在受伤时（n = 7）、受伤后第4天或第7天（分别为n = 7）注射生理盐水。实验II和III中的所有动物在受伤后第14天处死。使用张力计评估伤口抗张强度。分别使用生物测定法和酶联免疫吸附测定法检测从植入海绵中收集的伤口液中的肿瘤坏死因子-α和肿瘤坏死因子结合蛋白水平。通过Northern分析评估海绵肉芽肿中的胶原蛋白基因表达。通过测量羟脯氨酸含量对海绵中的胶原蛋白沉积进行定量。接受重复注射肿瘤坏死因子结合蛋白的动物的伤口明显较弱，平均伤口抗张强度为93.1g，而对照组为186.6g（p < 0.05）。在受伤后第0、4或7天接受单次注射肿瘤坏死因子结合蛋白的组的伤口抗张强度与各自的对照组无差异。任何肿瘤坏死因子结合蛋白组与其各自对照组之间海绵中的平均羟脯氨酸含量均无差异。胶原蛋白I和III表达的Northern分析也未显示差异。这些数据表明，持续全身给予肿瘤坏死因子结合蛋白会导致伤口明显较弱，而伤口胶原蛋白含量和胶原蛋白基因表达无相应差异。这表明在整个愈合过程中抑制肿瘤坏死因子-α会导致伤口质量受损，而胶原蛋白沉积没有定量改变。
