温和灸对大鼠慢性难愈性创面组织巨噬细胞及胶原表达的影响

目的:研究温和灸对慢性难愈性创面组织表面巨噬细胞及Ⅰ、Ⅲ型胶原含量的影响,探讨温和灸干预难愈性创面瘢痕形成的效果及可能作用机制。方法:在80只雄性SD大鼠中随机取72只,在其背部制造全层皮肤损伤开放性创面。予以肌肉注射氢化可的松琥珀酸钠,建立慢性难愈性创面动物模型,随机分为模型组、温灸组、TDP组,每组24只。不予任何干预的8只大鼠作为正常组。TDP组、温灸组取创面局部及双侧"肾俞"足三里"穴,分别以TDP照射或艾条温灸,每穴15min,每日1次。以HE染色法检测治疗第7、10、14天修复组织表面巨噬细胞数,以免疫组化法检测治疗第7、10、14天修复组织Ⅰ、Ⅲ型胶原的含量。结果:温灸组、TDP组、模型组创面修复组织巨噬细胞数均明显较正常组增多(P<0.05);在治疗第7天,温灸组表面巨噬细胞数多于模型组(P<0.05);在治疗第10天,温灸组、TDP组表面巨噬细胞数均多于模型组(P<0.05),且温灸组多于TDP组(P<0.05);在治疗第14天,温灸组、TDP组表面巨噬细胞数均较模型组减少(P<0.05),且温灸组少于TDP组(P<0.05)。大鼠创面修复组织Ⅰ、Ⅲ型胶原蛋白表达的平均吸光度,在治疗第7、14天,温灸组、TDP组均较模型组高(P<0.05,P<0.01);第14天温灸组高于TDP组(P<0.01)。温灸组、TDP组胶原蛋白Ⅰ/Ⅲ值均较模型组接近正常组;在第14天,温灸组优于TDP组(P<0.05)。结论:温和灸能促进创面愈合,且可降低瘢痕形成,其机制可能与调节不同时间内创面组织巨噬细胞数和Ⅰ、Ⅲ型胶原含量及其比值有关。

Effect of Mild Moxibustion on Macrophage Number and Collagen Expression of Raw Surface Tissue in Chronic Refractory Skin Ulcer Rats

Objective To observe the effect of mild moxibustion on the number of macrophages and contents of collagen Ⅰ and Ⅲ in the raw surface tissue in chronic skin ulcer rats so as to study its mechanism underlying promoting scar formation.Methods Eighty male SD rats were randomly divided into normal(n=8),model(n=24),TDP(n=24) and moxibustion(n=24) groups.Chronic refractory skin ulcer was established by making an open wound at the back and local intramuscular injection of hydrocortisone sodium succinate.For rats of the TDP and moxibustion groups,TDP irradiation or mild moxibustion was applied to the raw surface,bilateral "Shenshu"(BL 23) and "Zusanli"(ST 36) for 15 min,once daily for 7,10 and 14 days,respectively.The number of macrophages in the raw surface tissue was counted under light microscope after H.E.staining and collagen Ⅰ and Ⅲ expressions of the raw surface tissue were detected by immunohistochemistry.Results In comparison with the normal group,the numbers of macrophages in the raw surface tissue were increased significantly in the model group on day 7,10 and 14(P<0.05);while compared with the model group,the numbers of macrophages were increased further obviously in the moxibustion group on day 7 and 10 and in the TDP group on day 10 after the treatment(P<0.05).Compared with the model group,the numbers of macrophages in both TDP and moxibustion groups were down-regulated obviously(P<0.05).In regard to collagen Ⅰ and Ⅲ expression of the raw surface tissue,compared with the normal group,the collagen Ⅲ protein expression level was down-regulated markedly in the model group on the 7th day(P<0.01);whereas in comparison with the model group,the expression levels of collagen Ⅰ and Ⅲ were increased considerably in the TDP and moxibustion groups on day 7 and 14 after the treatment(P<0.05,P<0.01).The ratios of collagen Ⅰ/ Ⅲ expression were remarkably higher in the model group than in the normal group on day 7 and 14(P<0.05),and significantly lower in the TDP group on day 7 and 14 and in the moxibustion group on day 14 than in the model group(P<0.05,P<0.01).The effects of moxibustion were obviously superior to those of TDP in up-regulating macrophage number on day 10,up-regulating collagen Ⅰ and Ⅲ expressions on day 14,and down-regulating macrophage number on day 14 after the treatment(P<0.05,P<0.01).No significant differences were found between the TDP and moxibustion groups in up-regulating macrophage number,and collagen Ⅰ and Ⅲ protein expressions,and in down-regulating the ratios of collagen Ⅰ/Ⅲ expression on day 7 after the treatment(P>0.05).Conclusion Mild moxibustion can regulate the number of macrophages and strengthen the expression of collagen proteins in the raw surface tissue in the chronic skin ulcer rats,which may contribute to its effect in promoting wound healing and reducing scar formation.