MEBT/MEBO对慢性难愈合创面组织中CK10表达水平的影响

摘要

目的 通过对比观察不同治疗方案对大鼠创面组织内CK10 表达水平的影响, 进一步揭示皮肤再生医疗技术(moist exposed burn therapy/moist exposed burn ointment, MEBT/MEBO) 治疗慢性难愈合创面的部分作用机制.方法 将90 只SPF 级Wistar 大鼠随机分为空白组(18 只)、急创组(18 只)、慢创组(18 只)、 rb-bFGF组(18 只) 及MEBO 组(18 只), 其中空白组大鼠仅做备皮处理, 急创组大鼠建立急性创面模型, 慢创组、rb-bFGF 组及MEBO 组大鼠建立慢性难愈合创面模型, 且空白组大鼠局部皮肤予以生理盐水纱布湿敷, 急创组及慢创组大鼠创面予以生理盐水治疗, rb-bFGF 组大鼠创面予以重组牛碱性成纤维细胞生长因子(recombinant bovine basic fibroblast growth factor, rb-bFGF) 凝胶治疗, MEBO 组大鼠创面予以湿润烧伤膏(moist exposed burn ointment, MEBO) 治疗, 分别于治疗第3、7、14天对比观察5 组大鼠皮肤或创面组织的形态学变化及CK10 的表达水平.结果 (1) 治疗第3天, 空白组大鼠皮肤组织形态与正常皮肤组织形态无明显差异, 其他各组大鼠创面组织水肿明显, 可见大量炎性细胞浸润及少量成纤维细胞与新生毛细血管分布; 治疗第14天, 空白组大鼠皮肤组织形态与正常皮肤组织形态无明显差异, 慢创组大鼠创面组织内仍有少量炎性细胞浸润, 且有大量排列不规则的成纤维细胞分布, 而急创组、rb-bFGF 组及MEBO 组大鼠创面组织内可见排列整齐的新生毛细血管及完整的毛囊和皮脂腺等皮肤附属器, 组织结构接近于正常皮肤组织.(2) 治疗第3、7、14天, 空白组大鼠皮肤组织内 CK10 表达水平无明显变化, P >0.05, 差异无统计学意义; 而急创组、慢创组、rb-bFGF 组及MEBO 组大鼠创面组织内CK10 表达水平呈现逐渐升高的趋势, P 均<0.05, 差异具有统计学意义.(3) 治疗第7、14天, 急创组、 rb-bFGF 组和MEBO 组大鼠创面组织内CK10 表达水平均明显高于慢创组, P 均< 0.05, 差异具有统计学意义;治疗第3、7、14天, rb-bFGF 组和MEBO 组大鼠创面组织内CK10 表达水平均无明显差异, P 均> 0.05, 差异无统计学意义.结论 MEBT/MEBO 治疗慢性难愈合创面的疗效不亚于rb-bFGF 凝胶, 且改变表皮干细胞的外界生存环境, 促进表皮干细胞向正常表皮细胞增殖、分化, 可能是其促进慢性难愈合创面修复的作用机制.%Objective To observe and compare the influences of different treatment approaches on the expression levels of CK10 in rat wound tissues, and further reveal partially the action mechanism of MEBT/MEBO in treating chronic non-healing wounds. Methods Ninety SPF Wistar rats were randomly divided into blank group (18 rats), acute wound group (18 rats), chronic wound group (18 rats), rb-bFGF group (18 rats) and MEBO group (18 rats). Rats in the blank group were only given skin preparation, acute wounds were established in rats in the acute wound group, while chronic non-healing wounds were established in rats respectively in the chronic wound group, rb-bFGF group and MEBO group. The prepared local skin of rats in the blank group was dressed with normal saline soaked gauze, wounds in the acute wound group and chronic wound group were managed with normal saline, wounds in the rb-bFGF group were treated with recombinant bovine basic fibroblast growth factor (rb-bFGF) gel, and wounds in the MEBO group were treated with MEBO. The morphological changes of rat skin or wound tissues and the expression levels of CK10 were observed and compared in the five groups respectively on day 3, 7 and 14 of treatment. Results (1) On day 3 of treatment, the morphology of rat skin tissues in the blank group showed no obvious difference from that of the normal skin tissues. In contrast, the rat wound tissues in the other four groups were markedly edematous, with plenty of inflammatory cell infiltration, few fibroblasts and new capillaries visible. On day 14 of treatment, there was still no obvious morphology difference between the skin tissue in the blank group and the normal skin tissues. There was mild inflammatory cell infiltration in the wound tissues of the chronic wound group, with a large number of fibroblasts arranging disorderly, while in the wound tissues of the acute wound group, rb-bFGF group and MEBO group, there were well-arranged newly-grown capillaries and intact skin appendages including hair follicles and sebaceous glands, with basically the same tissue structure as the normal skin tissues. (2) On day 3, 7 and 14 of treatment, no significant change was observed on the expression level of CK10 in rat skin tissues in the blank group, and there was no statistically significant difference, P >0.05. In contrast, the expression of CK10 in rat wound tissues in the acute wound group, chronic wound group, rb-bFGF group and MEBO group all showed an increasing tendency, and the differences were statistically significant, all P <0.05. (3) On day 7 and 14 of treatment, the expression levels of CK10 in rat wound tissues in the acute wound group, rb-bFGF group and MEBO group were all obviously higher than that in the chronic wound group, and the differences were statistically significant, all P < 0.05. On day 3, 7 and 14, there was no statistically significant difference in the expression level of CK10 in rat wound tissues between the rb-bFGF group and MEBO group, all P >0.05. Conclusion The clinical efficacy of MEBT/MEBO in the treatment of chronic non-healing wounds is not inferior to that of rb-bFGF gel. Moreover, this approach can change the external living environment of epidermal stem cells and promote the proliferation of epidermal stem cells and their differentiation into normal dermal cells, which may be its action mechanism of facilitating the repair of chronic non-healing wounds.