大鼠全层皮肤缺损创面愈合过程中巨噬细胞浸润及表型研究  

Infiltration of macrophages and their phenotype in the healing process of full-thickness wound in rat

作　　者：牛轶雯[1] 缪明远 曹晓赞[1] 宋菲[1] 嵇晓芸[1] 董叫云[1] 陆树良[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院上海市烧伤研究所,上海市创面修复研究中心,200025 [2]上海交通大学医学院附属上海儿童医学中心

出　　处：《中华烧伤杂志》2014年第2期109-115,共7页Chinese Journal of Burns

基　　金：国家自然科学基金面上项目（81272111、81071568）;上海市卫生局青年科研项目（20134y085）

摘　　要：目的研究大鼠全层皮肤缺损创面愈合过程巾臣噬细胞的浸润及表型变化。方法取30只继康S19大鼠，按随机数字表法分为损伤组24只、对照组6只。损伤组大鼠于脊柱两侧用自制环钻及手术剪制成2个直径为11mm的全层皮肤缺损创面，致伤后即刻测量创面面积，每日碘伏消毒；对照组大鼠仪行麻醉脱毛处理。伤后1、3、7、13d，分刖取损伤组6只大鼠测量创面面积（计算创面愈合率）后处死。沿创缘切取创面组织达健康筋膜层，HE染色观察组织学表现，免疫组织化学染色观察纰织巾巨噬细胞表面标志物CD68表达，免疫荧光染色分别观察组织中CD68与诱导型一氧化氮合酶（iNOS）双阳性（I型曩噬细胞）、CD68与精氨酸酶1（Arg-1）双阳性（1／型巨噬细胞）表达情况，双抗体夹心ELISA法检测创面组织巾叮十扰素、TNF-α、IL4、IL-13、IL-10和IL-12的水平并计算IL-10／IL-12比值。对照组大鼠在与损伤组相同部位切取直径为11mm的全层正常皮肤组织，同前行组够5学及细胞因子检测。对数据行单因素方差分析或LSD-t检验。结果损伤组大鼠伤后创面逐渐缩小，伤后各时棚点创面愈合率总体比较差异有统计学意义（F=358．55，P〈0．01）。对照组大鼠皮肤组织形念未见异常：损伤组大鼠伤后1、3d，创面组织中炎性细胞明显浸润；伤后7、13d，可见明显血管腔结构、新乍胶原。对照组大鼠正常组织和损伤组大鼠伤后l、3、7、13d创面组织每200倍视野下的CD68阳性细胞数分别为（2．7±1．5）、（31．8±3．5）、（40．8±4．7）、（20．8±2．8）、（3．2±2．4）个（F=180．55，P〈0．01）。损伤组大鼠伤后1、3、7dCD68阳性细胞数明显高于对照组（，值分圳为18．8l、18．79、14．05，P值均小于0．01）。对照组大鼠正常组织巾未见CD68与iNOS双阳性或者CD68与Arg-1双阳性细胞。损伤组大鼠伤后1、3、7、1Objective To study the infiltration of macrophages and their phenotype in the healing process of full-thickness wound in rat. Methods Thirty healthy SD rats were divided into control group （ n = 6） and injury group （ n = 24） according to the random number table. Two round full-thickness skin defects （11 mm diameter） were created on both sides of dorsal spine of rats in injury group with surgical scissors and homemade trephine. After injury, wound area was measured immediately. The wounds were dis- infected with iodophor every day. Rats in control group received anesthesia and hair removal only. On post injury day （PID） 1 , 3, 7, and 13, respectively, 6 rats of injury group were sacrificed after the measurement of wound area （wound healing rate was calculated）. Wound samples were obtained by excision down to healthy fascia along wound edge. Histological study was done with HE staining. The expression of CD68 （the surface marker of macrophage） in the wound tissue was observed with immunohistochemieal staining. The double positive expressions of induced nitric oxide synthase （iNOS） plus CD68 （type I macrophage） and arginase 1 （Arg-1） plus CD68 （type II macrophage） were observed with immunofluoreseenee staining. The levels of interferon-＇y （IFN-γ） , TNF-α, IL-4, IL-13, IL-10, and IL-12 in wound tissue were assayed by double-antibody sandwich ELISA, and the ratio of IL-10/IL-12 was calculated. Full-thickness skin tis- sues （ 11 mm diameter） in rats of control group were excised at the same site as rats in injury group, and the histological observation and cytokines assay were performed as well. Data were processed with one-way analy- sis of variance or LSD- t test. Results Wound area of rats in injury group was gradually reduced after in- jury, and the overall difference of the wound healing rate on each PID was statistically significant （ F = 358.55, P 〈 0.01 ）. No abnormal appearance of skin tissue was observed in rats of control group. In injury group, infla

关 键 词：伤口愈合 巨噬细胞 表型 

分 类 号：R622.1[医药卫生—整形外科]