Si Zhongyi, Bhardwaj Rhanjit, Rosch Raphael, Mertens Peter Rene, Klosterhalfen Bernd, Klinge Uwe
Department of Surgery, the Department of Internal Medicine II, and the Institute of Pathology, Technical University of Aachen, Aachen, Germany.
Surgery. 2002 Mar;131(3):324-31. doi: 10.1067/msy.2002.121376.
Recent findings of an impaired protein ratio of type I to type III procollagen showed a disturbed collagen metabolism in incisional hernia development. We analyzed the type I and type III procollagen messenger RNA to investigate whether these findings represent the altered extracellular matrix or a primary defect at the transcriptional level.
We examined cultured skin fibroblasts of patients with incisional or recurrent incisional hernia in comparison with those without any previous incision (control) and those with a skin scar without clinical appearance of a hernia (scar). Immunohistochemical detection of a lowered protein ratio of type I and type III collagen in the hernia skin tissue leads to mRNA expression analysis. The procollagen mRNA and the ratio of type I to type III procollagen mRNA are detected by reverse transcriptase-polymerase chain reaction and Northern blot analysis, the collagens type I and III by Western blot analysis.
Reverse transcriptase-polymerase chain reaction revealed an increase of type I procollagen mRNA in the incisional and recurrent hernia (0.90 +/- 0.04 and 1.19 +/- 0.04, respectively) compared with stable scar (0.54 +/- 0.02) or healthy tissue (0.43 +/- 0.01). The obvious rise of type III procollagen mRNA to 4.13 +/- 0.04 for incisional, 6.02 +/- 0.03 for recurrent hernia, 2.29 +/- 0.04 for stable scar, and 1.72 +/- 0.03 for the healthy tissue showed a significantly decreased ratio of type I to type III procollagen mRNA in the hernia patients as compared with the controls (P <.01). By Western blot analysis, an increase of type I and type III collagen protein and a significant rise in the corresponding ratio in the recurrent hernia group were detected.
The altered synthesis of type I and type III collagen in cultured skin fibroblasts suggests a disorder of collagen metabolism, at least in patients with recurrent hernia. Hence, a basically impaired wound healing process is likely to contribute to the unsatisfactory results of incisional hernia repair.
近期研究发现,I型与III型前胶原的蛋白比例受损,这表明在切口疝形成过程中胶原蛋白代谢紊乱。我们分析了I型和III型前胶原信使核糖核酸,以研究这些发现是代表细胞外基质改变还是转录水平的原发性缺陷。
我们检查了切口疝或复发性切口疝患者的培养皮肤成纤维细胞,并与无既往手术切口的患者(对照组)以及有皮肤瘢痕但无疝临床表现的患者(瘢痕组)进行比较。通过免疫组织化学检测发现疝皮肤组织中I型和III型胶原蛋白的蛋白比例降低,进而进行信使核糖核酸表达分析。通过逆转录聚合酶链反应和Northern印迹分析检测前胶原信使核糖核酸以及I型与III型前胶原信使核糖核酸的比例,通过蛋白质印迹分析检测I型和III型胶原蛋白。
逆转录聚合酶链反应显示,与稳定瘢痕(0.54±0.02)或健康组织(0.43±0.01)相比,切口疝和复发性疝中I型前胶原信使核糖核酸增加(分别为0.90±0.04和1.19±0.04)。III型前胶原信使核糖核酸在切口疝中明显升高至4.13±0.04,复发性疝中为6.02±0.03,稳定瘢痕中为2.29±0.04,健康组织中为1.72±0.03,这表明与对照组相比,疝患者中I型与III型前胶原信使核糖核酸的比例显著降低(P<0.01)。通过蛋白质印迹分析,在复发性疝组中检测到I型和III型胶原蛋白增加,且相应比例显著升高。
培养的皮肤成纤维细胞中I型和III型胶原蛋白合成改变表明胶原蛋白代谢紊乱,至少在复发性疝患者中如此。因此,伤口愈合过程基本受损可能是切口疝修复效果不理想的原因之一。