[1]韩秀芳,曾婷婷,段文丽,等.基于细胞衍生因子1/趋化因子受体4通路探究巨噬细胞极化在象皮生肌膏治疗大鼠肛瘘术后模型创面中的作用实验研究[J].陕西医学杂志,2025,54(3):296-300,306.[doi:DOI:10.3969/j.issn.1000-7377.2025.03.002]
　HAN Xiufang,ZENG Tingting,DUAN Wenli,et al.Role of macrophage polarization in wound healing of rats anal fistula model treated with Xiangpi Shengji ointment based on SDF-1/CXCR4 pathway[J].,2025,54(3):296-300,306.[doi:DOI:10.3969/j.issn.1000-7377.2025.03.002]
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基于细胞衍生因子1/趋化因子受体4通路探究巨噬细胞极化在象皮生肌膏治疗大鼠肛瘘术后模型创面中的作用实验研究
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
54
期数:
2025年3期
页码:
296-300,306
栏目:
基础研究
出版日期:
2025-03-05

文章信息/Info

Title:
Role of macrophage polarization in wound healing of rats anal fistula model treated with Xiangpi Shengji ointment based on SDF-1/CXCR4 pathway
作者:
韩秀芳; 曾婷婷; 段文丽; 周 莉
(湖南中医药大学第一附属医院肛肠科,湖南 长沙 410007)
Author(s):
HAN Xiufang; ZENG Tingting; DUAN Wenli; ZHOU Li
(Department of Colorectal Medicine,the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine,Changsha 410007,China)
关键词:
瘘;  象皮生肌膏;  细胞衍生因子1;  趋化因子受体4;  巨噬细胞;  创面修复;  大鼠
Keywords:
Anal fistula;  Xiangpi Shengji ointment;  Stromal cell-derived factor 1;  C-X-C motif chemokine receptor 4;  Macrophages;  Wound healing;  Rats
分类号:
R 285.5
DOI:
DOI:10.3969/j.issn.1000-7377.2025.03.002
文献标志码:
A
摘要:
目的:基于基质细胞衍生因子1(SDF-1)/趋化因子受体4(CXCR4)通路探讨巨噬细胞极化在象皮生肌膏治疗大鼠肛瘘术后模型创面中的作用。方法:将10只SPF级健康成年SD大鼠(对照组)、10只SDF-1基因特异性敲除大鼠(SDF-1-/- 组)和10只SDF-1基因过表达大鼠(SDF-1过表达组)通过钢丝引流进行肛瘘造模,之后对大鼠实施肛瘘瘘管切开术建立肛瘘术后创面模型。各组大鼠统一给予象皮生肌膏治疗10 d。每组大鼠分别在治疗第0、5、10天采用常规面积检测法测量创面面积。Wetsern blot和实时荧光定量PCR(RT-qPCR)检测各组大鼠肉芽组织中SDF-1、CXCR4蛋白和mRNA表达水平。免疫荧光染色(IF)分析给药第10天各组大鼠肉芽组织中白细胞介素-6(IL-6)表达情况。流式细胞术分析各组大鼠肉芽组织中巨噬细胞极化情况。免疫组织化学染色(IHC)分析蛋白阳性表达情况。结果:在治疗第5、10天,SDF-1-/- 组大鼠创面愈合率均低于对照组大鼠,SDF-1过表达组大鼠创面愈合率高于SDF-1-/- 组和对照组(均P<0.05)。SDF-1过表达组大鼠创面基本愈合,对照组大鼠创面愈合情况良好,SDF-1-/- 组大鼠创面愈合情况最差。IF分析显示,SDF-1-/- 组大鼠肉芽组织中IL-6表达水平高于对照组,SDF-1过表达IL-6表达水平低于SDF-1-/- 组和对照组(均P<0.05)。流式细胞术分析表明,SDF-1-/- 组大鼠肉芽组织中巨噬细胞M1/M2比例高于对照组,SDF-1过表达组M1/M2比例低于SDF-1-/- 组和对照组(均P<0.05)。Western blot和RT-qPCR结果显示,SDF-1-/- 组大鼠肉芽组织中SDF-1、CXCR4蛋白和mRNA表达水平低于对照组,SDF-1过表达组SDF-1、CXCR4蛋白和mRNA高于SDF-1-/- 组和对照组(均P<0.05)。IHC染色分析表明,SDF-1-/- 组大鼠肛瘘创面肉芽组织内,M2型巨噬细胞标志物CD206与CXCR4同步降低,而在对照组和SDF-1过表达组均呈高水平表达。结论:SDF-1/CXCR4通路可以通过抑制炎症因子释放和诱导巨噬细胞M2极化抑制炎症反应,进而增强象皮生肌膏对肛瘘创面的修复作用。
Abstract:
Objective:To investigate the role of macrophage polarization in wound healing of rats anal fistula model treated with Xiangpi Shengji ointment based on stromal cell-derived factor 1(SDF-1)/C-X-C motif chemokine receptor 4(CXCR4)pathway.Methods:Anal fistula modeling was performed in 10 SPF healthy adult SD rats(control group),10 SDF-1 gene-specific knockout rats(SDF-1-/- group)and 10 SDF-1 gene-overexpressing rats(SDF-1 overexpressing group)by steel wire drainage,and then the wound surface model of anal fistula was established by incision of anal fistula.The rats in each group were treated with Xiangpi Shengji ointment for 10 days.The wound area of each group was measured by conventional area detection on the 0,5 and 10 days of treatment,respectively.Wetsern blot and RT-qPCR were used to detect the expression levels of SDF-1 and CXCR4 protein and mRNA in granulation tissues of each group.Immunofluorescence staining(IF)was used to analyze the expression of IL-6 in granulation tissue of each group on the 10th day of administration.Flow cytometry was used to analyze the polarization of macrophages in granulation tissue of each group.The positive expression of protein was analyzed by immunohistochemical staining(IHC).Results:On the 5th and 10th day of treatment,the wound healing rate of SDF-1-/- group was lower than that of control group,and the wound healing rate of SDF-1 overexpression group was higher than that of SDF-1-/- group and control group(all P<0.05).The wounds of the SDF-1 overexpression group were basically healed,the wounds of the control group were good,and the wounds of the SDF-1-/- group were the worst.IF analysis showed that the expression level of IL-6 in granulation tissue in SDF-1-/- group was higher than that in control group,and the expression level of IL-6 in SDF-1 overexpression group was lower than that in SDF-1-/- group and control group(all P<0.05).Flow cytometry analysis showed that the M1/M2 ratio of macrophages in granulation tissue in SDF-1-/- group was higher than that in control group,and the M1/M2 ratio in SDF-1 overexpression group was lower than that in SDF-1-/- group and control group(all P<0.05).Western blot and RT-qPCR results showed that the expression levels of SDF-1 and CXCR4 protein and mRNA in granulation tissue of SDF-1-/- group were lower than those of control group,and the protein and mRNA of SDF-1 and CXCR4 in SDF-1 overexpression group were higher than those in SDF-1-/- group and control group(all P<0.05).IHC staining analysis showed that the M2-type macrophage marker CD206 and CXCR4 in the anal fistula wound granulation tissue in SDF-1-/- group decreased simultaneously,while the expression levels of them were high in both the control group and the SDF-1 overexpression group.Conclusion:The expression of SDF-1/CXCR4 pathway can inhibit the inflammatory response by inhibiting the release of inflammatory factors and inducing macrophage M2 polarization,so as to enhance the repair effect of Qiangpi Shengji paste on anal fistula wound.

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备注/Memo

备注/Memo:
[基金项目]湖南省卫生健康委员会卫生科研课题(W20242016)
更新日期/Last Update: 2025-03-05