创缘铁过载的临床样本验证与铁过载对小鼠创面愈合的影响  

Clinical sample validation of iron overload at the wound edge and the effect of iron overload on wound healing in mice

作　　者：刘甜甜 李明[1,2] 朱含汀[1,2] 倪涛 彭银波[1,2] 方勇[1,2] Liu Tiantian;Li Ming;Zhu Hanting;Ni Tao;Peng Yinbo;Fang Yong(Department of Plastics and Reconstructive Surgery,Shanghai Ninth People′s Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200011,China;Institute of Trauma,Shanghai Jiao Tong University School of Medicine,Shanghai 200011,China)

机构地区：[1]上海交通大学医学院附属第九人民医院整复外科,200011 [2]上海交通大学医学院创伤医学研究所,200011

出　　处：《中华损伤与修复杂志（电子版）》2022年第6期475-481,共7页Chinese Journal of Injury Repair and Wound Healing(Electronic Edition)

基　　金：上海市科委振龙头项目(shslczdzk0091);上海交通大学医工(理)交叉基金(YG2019QNA15);上海市转化医学协同创新中心项目(TM202107);中华医学会生长因子振兴计划(SZYZ-TR-13);上海市科委项目(22MC1940300)。

摘　　要：目的创缘铁过载的临床样本验证与探讨铁过载对小鼠创面愈合的影响。方法(1)收集符合入选标准的2021年1月至2022年2月上海交通大学医学院附属第九人民医院门急诊及住院患者各期创面清创时的创缘组织共36份,按创面形成时间分为3组,分别为<1个月组、1~3个月组、>3个月组,每组12份。观察3组创缘色素沉着情况。取距离创缘前后各5 mm范围内的皮肤全层组织,通过电感耦合等离子体发射光谱检测创缘铁离子含量,苏木精-伊红染色及普鲁士蓝染色观察创缘组织炎症细胞分布与铁沉积部位及铁沉积范围。(2)将48只6~8周龄无特定病原体级雄性C57BL/6小鼠按随机数字表法分为对照组与铁过载组,每组24只。铁过载组小鼠腹腔注射于避光条件下配置而成含铁量为0.1 mg/mL的柠檬酸铁铵溶液,每次注射2 mL/只,每间隔3 d进行1次注射,共注射8次。对照组小鼠腹腔注射0.9%氯化钠溶液,注射含量及注射次数同铁过载组。注射结束后1 d,2组小鼠1%戊巴比妥钠腹腔注射(5μL/g)麻醉,电动剃毛刀剃除小鼠背部区域毛发,在背部建立1个1.5 cm×1.5 cm的全层创面。以建立创面当日定为造模成功即刻。造模成功后3、7、14 d,观察2组小鼠创缘色素沉着与创面愈合情况,并比较小鼠创面愈合率;2组造模成功即刻,造模成功后3、7、14 d各取6只小鼠脱颈处死并切去小鼠全层皮肤创缘组织,检测铁离子含量;苏木精-伊红染色与普鲁士蓝染色观察创缘组织炎症细胞分布与铁沉积部位及铁沉积范围。数据行单因素方差分析和LSD-t检验。结果(1)<1个月组创缘色红伴随局部炎症状态,随着创面形成时间的延长,1~3个月组、>3个月组创缘表现出较为明显的色素沉着;<1个月组、1~3个月组、>3个月组创缘铁离子含量分别为(35.24±2.82)、(70.02±6.16)、(106.90±9.62)μg/g,3组总体比较差异有统计学意义(F=27.82,P<0.05)。苏木精-伊红染色可Objective To verify the clinical samples of iron overload at the wound edge and investigate the effect of iron overload on wound healing in mice.Methods(1)A total of 36 wound marginal tissues from outpatients and inpatients who met the criteria in Shanghai Ninth People′s Hospital,Shanghai Jiaotong University School of Medicine at all stages of wound debridement from January 2021 to February 2022 were collected and divided into 3 groups according to the time of wound formation,<1 month group 1 to 3 months group and>3 months group,with 12 in each group.The pigmentation of the trabecular margin was observed in the three groups.The whole skin tissue within 5 mm from the front and back of the wound edge was taken,the iron ion content of the wound edge was detected by inductively coupled plasma emission spectroscopy,and the distribution of inflammatory cells in the tissue of the lesion was observed by hematoxylin-eosin staining and Prussian blue staining with the site and extent of iron deposition.(2)Forty-eight 6-8 week-old male C57BL/6 mice without specific pathogens were divided into control group and iron overload group according to the random number table method,with 24 mice in each group.The mice in the iron-overloaded group were injected intraperitoneally with ferric ammonium citrate solution at 0.1 mg/mL under light-protected conditions at an interval of every 3 d for 8 injections.The mice in the control group were injected intraperitoneally with 0.9%sodium chloride solution,and the injection content and number of injections were the same as those of the iron overload group.One day after the end of injection,mice in the 2 groups were anesthetized with 1%sodium pentobarbital intraperitoneally(5μL/g),and the hair in the dorsal region of the mice was shaved with an electric shaver to create a 1.5 cm×1.5 cm full-length wound on the back.The day when the wound was created was defined as the immediately successful modeling.At 3,7,and 14 d after successful modeling,the pigmentation and healing of the wound margins

关 键 词：创伤和损伤 小鼠 伤口愈合 铁过载 

分 类 号：R644[医药卫生—外科学]