摘要
目的探讨去铁胺对深部组织损伤(DTI)小鼠巨噬细胞极化和创面愈合的影响及其作用机制。方法采用实验研究方法。取54只6~8周龄雄性C57BL/6J小鼠,按随机数字表法分为DTI对照组、2 mg/mL去铁胺组、20 mg/mL去铁胺组,每组18只。采用磁铁压迫法在小鼠背部制造DTI,从伤后1 d开始,每隔1 d在创缘皮下注射100µL生理盐水或相应质量浓度的去铁胺溶液,直至取材;另取6只不进行任何处理的小鼠为正常对照组。取3组DTI小鼠,每组6只,于伤后3、7、14 d观察创面变化并计算创面愈合率。于其他组伤后3 d取正常对照组小鼠正常皮肤组织(下同)和其余3组小鼠伤后7、14 d创面组织,行苏木精-伊红(HE)染色观察组织形态。取正常对照组小鼠正常皮肤组织和其余3组小鼠伤后7 d创面组织,行免疫组织化学染色观测CD206和CD11c阳性面积百分比,分别采用实时荧光定量反转录PCR法及蛋白质印迹法检测CD206、CD11c和诱导型一氧化氮合酶(iNOS)的mRNA及蛋白表达。取正常对照组小鼠正常皮肤组织和DTI对照组、20 mg/mL去铁胺组小鼠伤后3、7、14 d创面组织,采用蛋白质印迹法检测信号转导及转录活化因子3(STAT3)和白细胞介素10(IL-10)蛋白表达。以上实验各组各时间点样本数均为6。取RAW264.7细胞,分为进行相应处理的50μmol/L去铁胺组、100μmol/L去铁胺组、200μmol/L去铁胺组和空白对照组,每组3孔,于培养48 h,采用流式细胞仪检测CD206和CD86阳性细胞百分比。数据分析采用重复测量方差分析、单因素方差分析和LSD检验。结果伤后7 d,2 mg/mL去铁胺组和20 mg/mL去铁胺组小鼠创面愈合率分别为(17.7±3.7)%、(21.5±5.0)%,均明显高于DTI对照组的(5.1±2.3)%(P<0.01);伤后14 d,2 mg/mL去铁胺组和20 mg/mL去铁胺组小鼠创面愈合率分别为(51.1±3.8)%、(57.4±4.4)%,均明显高于DTI对照组的(25.2±3.8)%(P<0.01)。HE染色可见,正常对照组小鼠正常皮肤组织层次清�
Objective To investigate the effects of deferoxamine on macrophage polarization and wound healing in mice with deep tissue injury(DTI)and its mechanism.Methods The experimental research methods were adopted.Fifty-four male C57BL/6J mice of 6-8 weeks old were divided into DTI control group,2 mg/mL deferoxamine group,and 20 mg/mL deferoxamine group according to random number table,with 18 mice in each group.DTI was established on the back of mice by magnet compression method.From post injury day(PID)1,mice were injected subcutaneously with 100µL normal saline or the corresponding mass concentration of deferoxamine solution every other day at the wound edge until the samples were collected.Another 6 mice without any treatment were selected as normal control group.Six mice in each of the three DTI groups were collected on PID 3,7,and 14 to observe the wound changes and calculate the wound healing rate.Normal skin tissue of mice in normal control group was collected on PID 3 in other groups(the same below)and wound tissue of mice in the other three groups on PID 7 and 14 was collected for hematoxylin-eosin(HE)staining to observe the tissue morphology.Normal skin tissue of mice in normal control group and wound tissue of mice in the other three groups on PID 7 were collected,and the percentages of CD206 and CD11c positive area were observed and measured by immunohistochemical staining,and the mRNA and protein expressions of CD206,CD11c,and inducible nitric oxide synthase(iNOS)were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction and Western blotting,respectively.Normal skin tissue of mice in normal control group and wound tissue of mice in DTI control group and 20 mg/mL deferoxamine group were collected on PID 3,7,and 14,and the protein expressions of signal transducer and activator of transcription 3(STAT3)and interleukin-10(IL-10)were detected by Western blotting.The sample number in each group at each time point in the above experiments.The RAW264.7 cells were divided
作者
山慧
张子锐
王晓莹
侯佳雨
张菊
Shan Hui;Zhang Zirui;Wang Xiaoying;Hou Jiayu;Zhang Ju(School of Nursing,Qingdao University,Qingdao 266071,China;Department of Intensive Care Medicine,Affiliated Hospital of Qingdao University,Qingdao 266555,China)
出处
《中华烧伤与创面修复杂志》
CAS
CSCD
北大核心
2022年第8期767-777,共11页
Chinese Journal of Burns And Wounds
基金
国家自然科学基金青年科学基金项目(81701838)
中国博士后科学基金项目(2018M632628)。
关键词
伤口愈合
去铁胺
巨噬细胞极化
深部组织损伤
Wound healing
Deferoxamine
Macrophage polarization
Deep tissue injury
