紫朱软膏对急性创面愈合过程中IL-1β、MMP-9和TGF-β的作用及自噬相关性研究

Effect of Zizhu Ointment on IL-1β, MMP-9 and TGF-β during Acute Wound Healing and Correlation of Autophagy and Would Healing

目的:观察紫朱软膏对急性创面愈合的疗效、作用机制及自噬和创面愈合的相互关系。方法:SD大鼠背部急性皮肤创伤形成后随机分为两组,治疗组创面外用紫朱软膏,对照组给予生理盐水外敷,比较两组创伤后4 d、7 d、14 d和21 d的创面愈合率,HE染色观察创面组织形态,透射电镜观察各时相点创面自噬体,RT-PCR法检测自噬标志基因Beclin-1、LC3的表达及变化;同时观察创面愈合各阶段IL-1β、MMP-9和TGF-β的表达变化,在mRNA水平上分析其与自噬基因的关联性。结果:治疗组与对照组相比,愈合率高,愈合时间短,再上皮化速度快,电镜下可见自噬体减少,LC3、Beclin-1表达明显减弱;与对照组比较,治疗组创面IL-1β表达降低,与LC3显著正相关,MMP-9和TGF-β表达升高,与LC3显著负相关。结论:紫朱软膏在急性创伤修复过程中可能通过抑制细胞的自噬活性,下调IL-1β的表达减少炎症反应,上调MMP-9和TGF-β的表达,促进血管新生、组织纤维化,从而达到创面愈合的目的。

Objective:To observe the effect and action mechanism of Zizhu Ointment on acute wound healing,and to explore the relationship between autophagy and wound healing.Methods:SD rat model with acute skin injury were randomly divided into the control group and the treatment group.The treatment group was treated with external application of Zizhu Ointment,and the control group was given external application of normal saline.The wound healing rate of the two groups was compared on 4 d,7 d,14 d and 21 d after trauma.The morphology of wounds was observed by HE staining,and the autophagosomes of wounds were observed by transmission electron microscopy.The expressions of autophagy marker genes,Beclin-1 and LC3,were detected by RT-PCR.At the same time,the expressions of IL-1β,MMP-9 and TGF-βwere observed during the different wound healing stages,and the correlation with autophagy gene was analyzed at mRNA level.Results:The treatment group had a higher healing rate,a shorter healing time,and a faster epithelialization rate when compared to the control group.Under the electron microscope,autophagosomes of the treatment group were reduced,and the expressions of LC3 and Beclin-1 were significantly decreased.The expression of IL-1βwas significantly lower in the treatment group than that in the control group,which was significantly positively correlated with LC3.The expressions of MMP-9 and TGF-βwere higher in the treatment group than those in the control group,which were significantly negatively correlated with LC3.Conclusion:During acute wound repair,Zizhu Ointment may inhibit the autophagy activity of cells,down-regulate the expression of IL-1βto reduce the inflammatory response and up-regulate the expressions of MMP-9 and TGF-βto promote angiogenesis and tissue fibrosis,thus to achieve wound healing.