丹黄散对糖尿病大鼠创面愈合及新生皮肤中CK14、CK15表达的影响研究

Effect of Danhuang powder on wound healing and expression of CK14 and CK15 in regenerative skin in diabetic rats

目的探讨丹黄散对糖尿病大鼠创面愈合及新生皮肤中细胞角蛋白14(CK14)、细胞角蛋白15(CK15)表达的影响。方法链脲佐菌素(STZ)诱导糖尿病模型大鼠成功后,将实验大鼠随机分成丹黄散组、西药组、模型对照组和空白对照组,每组20只。在所有实验动物背部两侧制作直径约1 cm的圆形全层皮肤缺损创面,从造模当日开始,丹黄散组清创后外撒丹黄散后无菌棉纱覆盖;西药组清创后用山莨菪碱、庆大霉素和胰岛素的无菌棉纱外敷,空白对照组及模型对照组清创后无菌棉纱覆盖。分别于造模3 d、7 d、14 d、21 d后观察和记录各组创面未愈合面积,计算各时间点创面愈合率;取各组大鼠创缘新生皮肤组织进行HE染色,观察创缘组织修复情况及局部炎症反应;采用免疫组化法检测各组大鼠创缘组织中CK14和CK15蛋白表达情况。结果丹黄散组大鼠3 d时可见少量肉芽组织开始形成,21 d时创面基本愈合结痂,无死亡大鼠;西药组大鼠7 d时可见肉芽组织出现,到21 d表皮覆盖大部分创面,无死亡大鼠;模型对照组大鼠创面愈合缓慢,7 d、12 d各死亡1只,21 d时创面大多未愈合;空白对照组大鼠3 d时即可见新鲜肉芽组织生成,18 d时死亡1只,21 d时创面基本愈合。西药组、丹黄散组大鼠各时间点创面愈合率及创面组织中CK14、CK15表达水平均明显高于模型对照组(P均<0.05);丹黄散组大鼠3 d、7 d时创面愈合率及创面组织中CK14、CK15表达水平与西药组比较差异无统计学意义(P均>0.05),14 d、21 d时均明显高于西药组(P均<0.05)。结论丹黄散可有效促进糖尿病大鼠皮肤溃疡创面愈合,机制可能与有效调控CK14、CK15表达有关。

Objective It is to explore the effect of Danhuang powder on wound healing and expression of cytokeratin 14(CK14)and cytokeratin 15(CK15)in regenerative skin in diabetic rats.Methods The diabetes models of rats were induced by streptozotocin(STZ)and were randomly divided into Danhuang powder group,western medicine group,model control group and blank control group,20 rats in each group.A round full-thickness skin defect wound with a diameter of about 1 cm was made on both sides of the back of all experimental animals.From the day of modeling,the Danhuang San group was covered with sterile cotton after the Danhuang powder was debrided.The western medicine group was covered externally with sterile cotton yarns of anisodamine,gentamicin and insulin after debridement,the blank control group and model control group were covered with sterile cotton yarns after debridement.The unhealed area of the wounds in each group in 3 d,7 d,14 d,and 21 d after modeling were observed and recorded,the wound healing rate at each time point was calculated.The new skin tissue of the wound margin of each group was taken for HE staining to observe the repair status of the wound margin tissue and local inflammatory response;the expression of CK14 and CK15 proteins in wound tissue of rats in each group was detected by immunohistochemistry.Results A small amount of granulation tissue started to form in the rats of Danhuangsan group at 3 days,and the wounds healed and crusted on the 21st day,there was no dead rat.Rats in the western medicine group showed granulation tissues at 7 days,and the epidermis covered most of the wounds at 21 days,there was no dead rat.The wounds healed slowly in model control group,one rat died at 7 d and 12 d,most of the wounds did not heal at 21 d;the formation of fresh granulation tissue at 3 d was found in the rats of blank control group rats,1 rat died at 18d,and the wound was basically healed at 21 d.The wound healing rate and the expression levels of CK14 and CK15 in the wound tissues of the rats in the western medicine group and the Danhuangsan group were significantly higher than those in the model control group(P<0.05).The rates and expression levels of CK14 and CK15 in wound tissue were not significantly different from those of the western medicine group(both P>0.05),and were significantly higher than those of the western medicine group at 14d and 21d(both P<0.05).Conclusion Danhuang powder can effectively promote the healing of skin ulcer wounds in diabetic rats,and the mechanism may be related to the effective regulation of CK14 and CK15 expression.