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Impaired balance of type I and type III procollagen mRNA in cultured fibroblasts of patients with incisional hernia.

机译:切口疝患者培养的成纤维细胞中I型和III型前胶原mRNA的平衡受损。

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BACKGROUND: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.
机译:背景:I型与III型胶原蛋白原蛋白比例受损的最新发现表明,切口疝的发展中胶原蛋白的代谢受到了干扰。我们分析了I型和III型前胶原信使RNA,以调查这些发现是否代表改变的细胞外基质或转录水平的主要缺陷。方法:我们比较了有切口或复发性切口疝患者的培养的皮肤成纤维细胞与没有任何切口(对照)的患者和有皮肤疤痕但没有临床疝的患者(瘢痕)的比较。免疫组织化学检测疝皮肤组织中I型和III型胶原蛋白的蛋白质比率降低,从而导致mRNA表达分析。通过逆转录酶-聚合酶链反应和Northern印迹分析检测原胶原mRNA和I型与III型胶原原mRNA的比例,通过Western印迹分析检测I型和III型胶原。结果:逆转录酶-聚合酶链反应显示,与稳定的疤痕(0.54 +/- 0.02)或健康者相比,切口和复发性疝的I型胶原蛋白mRNA增加(分别为0.90 +/- 0.04和1.19 +/- 0.04)组织(0.43 +/- 0.01)。 III型前胶原mRNA的明显增加为切入为4.13 +/- 0.04,复发性疝为6.02 +/- 0.03,稳定疤痕为2.29 +/- 0.04和健康组织的1.72 +/- 0.03显示比率显着降低与对照组相比,疝患者中I型至III型前胶原mRNA的变化(P <.01)。通过蛋白质印迹分析,发现复发性疝气组中I型和III型胶原蛋白增加,相应比例显着增加。结论:培养的皮肤成纤维细胞中I型和III型胶原蛋白合成的改变提示胶原代谢异常,至少在复发性疝患者中。因此,基本受损的伤口愈合过程很可能导致切开疝修复的效果不理想。

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