摘要
目的探索水溶性积雪草酸葡糖胺盐(AAGS,凝胶形式)对大鼠皮肤缺损创面的作用及其机制。方法选取14只5周龄SD大鼠,每只大鼠背部两侧均建立相同的创面模型,一侧设为实验组,一侧设为对照组。实验组每天外用AAGS凝胶(30 mg/mL溶于PBS);对照组每天外用凝胶敷料(清得佳)。分别在第0、3、7、10和14天进行大体观并测量创面面积。分别在第3天(n=4)、第7天(n=4)和第14天(n=6)取材,HE染色观察并测定表皮厚度和表皮嵴数量,半定量分析炎性细胞聚集情况;免疫组化染色观察创面愈合处巨噬细胞向M1型、M2型极化的情况。小鼠单核巨噬细胞(RAW 264.7)培养过程中加入梯度浓度AAGS,Q-PCR检测M1、M2型巨噬细胞标记基因的表达情况。结果第0、3、7、10和14天大体观可见实验组相对愈合较快,定量分析显示在第10天实验组创面面积明显小于对照组(P<0.05)。实验组表皮嵴数量明显多于对照组(第14天,P<0.05),表皮厚度明显低于对照组(第14天,P<0.05)。实验组早期(第3天和第7天)的炎症细胞数量明显多于对照组(P<0.05)。此外,经过14 d的治疗,实验组巨噬细胞向M2型极化,体外细胞学研究也证实AAGS可诱导巨噬细胞向M2型的极化。结论AAGS凝胶具有增强局部表皮结构再生的潜力,可作为局部使用药物来促进创面愈合。AAGS凝胶在创面早期促进炎症过程以增强新生组织形成,并在创面晚期诱导巨噬细胞M2型极化以达到重塑和再生皮肤结构的作用。
Objective To explore the effect and mechanism of water soluble asiatic acid glucosamine salt(AAGS,gel form)on skin resection wounds in rats.Methods Fourteen 5-week-old SD rats were selected,and wound models were established on both sides of the back of each rat.One side was set as the experimental group,and the other was set as the control group.The experimental group applied AAGS gel(30 mg/mL in PBS)externally every day;the control group applied Intrasite gel externally every day.Gross observation and wound area measurement were performed on day 0,3,7,10 and 14,respectively.Animals were sacrificed and harvested on day 3(n=4),day 7(n=4),and day 14(n=6),the thickness of epidermis and the number of epidermal ridge were measured by HE staining,and the inflammatory cell aggregation was analyzed semi-quantitatively.Immunohistochemical staining was used to observe the polarization of M1 and M2 macrophages at the wound healing site.Mouse mononuclear macrophages(RAW 264.7)were cultured with gradient AAGS and Q-PCR was used to detect the expression of M1 and M2 macrophage marker genes.Results The wound healing on day 0,3,7,10 and 14 showed that the experimental group healed relatively quickly.Quantitative analysis showed that the wound area on the 10th day was significantly smaller than that of the control group(P<0.05).The number of epidermal ridge in the experimental group was significantly higher than that in the control group(Day 14,P<0.05),and the thickness of the epidermis was significantly lower than that in the control group(Day 14,P<0.05).The number of inflammatory cells at the early stage(Day 3 and 7)in the experimental group was significantly higher than that in the control group(P<0.05).In addition,after 14 days of treatment,the macrophages of the experimental group were polarized to M2 type,and in vitro cytological studies also confirmed that AAGS could induce M2 type polarization of macrophages.Conclusion AAGS gel has the potential to enhance the regeneration of local epidermal structures and can be used as
作者
刘冰滢
王文波
黄佳
高振
武晓莉
刘伟
LIU Bingying;WANG Wenbo;HUANG Jia;GAO Zhen;WU Xiaoli;LIU Wei(Plastic Surgery Research Institute,Weifang Medical University,Weifang 261042,China;Department of Plastic and Reconstructive Surgery,Shanghai Ninth People′s Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200011,China;Shanghai Key Laboratory of Tissue Engineering,Shanghai 200011,China)
出处
《组织工程与重建外科杂志》
2020年第2期112-118,共7页
Journal of Tissue Engineering and Reconstructive Surgery
关键词
积雪草酸葡糖胺盐
创面愈合
巨噬细胞极化
炎症
Asiatic acid glucosamine salt
Wound healing
Macrophage polarization
Wound inflammation
