使用牛分枝杆菌减毒株建立大鼠结核性创面模型的可靠性探究

周克强; 苏映军; 贾赤宇

doi:10.3760/cma.j.cn501120-20200531-00290

使用牛分枝杆菌减毒株建立大鼠结核性创面模型的可靠性探究

doi: 10.3760/cma.j.cn501120-20200531-00290

1.
山西医科大学第二临床医学院整形外科，太原　　030001
2.
西安国际医学中心整形外科医院　　710118
3.
厦门大学附属翔安医院烧伤整形与创面修复科　　361102

基金项目:

福建省自然科学基金 2019J01011

厦门大学附属翔安医院科研启动经费 PM201809170010

详细信息

通讯作者:
贾赤宇，Email：jiachiyu@qq.com

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出版历程
- 收稿日期: 2020-05-31

Reliability study on establishing a rat tuberculosis wound model using Bacillus Calmette-Guérin

1.
Department of Plastic Surgery,Second Clinical Medical College of Shanxi Medical University,Taiyuan 030001, China
2.
Plastic Surgery Hospital, Xi'an International Medical Center, Xi'an 710118, China
3.
Department of Burns and Plastic SurgeryWound Repair Surgery, Xiang'an Hospital of Xiamen University, Xiamen 361102, China

Funds:

Natural Science Foundation of Fujian Province of China 2019J01011

Starting Package of Xiang'an Hospital of Xiamen University PM201809170010

More Information

Corresponding author: Jia Chiyu, Email: jiachiyu@qq.com

摘要

摘要: 目的探讨通过注射牛分枝杆菌减毒株（BCG）建立大鼠结核性创面模型的可靠性。方法采用实验研究方法。将15只6周龄雄性SD大鼠按照随机数字表法分为正常对照组和感染组，其中正常对照组3只、感染组12只。感染组大鼠在背部皮下注射弗氏完全佐剂3周后皮下注射BCG菌液，建立大鼠结核性创面模型；正常对照组不做任何处理。于感染第8、15、32、43天，大体观察感染组大鼠注射部位皮肤情况；于前述时间点分别取感染组3只大鼠注射部位皮肤组织，另取正常对照组大鼠对应部位皮肤组织，行苏木精-伊红染色，观察细胞排列、坏死及炎症等情况。对感染第43天感染组大鼠注射部位皮肤组织行抗酸染色，观察细菌分布情况。结果感染第8、15、32、43天，感染组大鼠注射部位皮肤逐渐形成结核性创面病灶，病变组织细胞排列杂乱或呈同心圆状，坏死细胞及肉芽肿数量逐渐增多；正常对照组大鼠对应部位的皮肤组织细胞排列规律，未见炎症细胞浸润。感染第43天，感染组大鼠注射部位皮肤组织中可见大量杆状细菌。结论使用BCG建立的大鼠结核性创面模型稳定可靠，可以满足实验要求。
- 模型，动物 /
- 结核，皮肤 /
- 大鼠 /
- 牛分枝杆菌减毒株
Abstract: Objective To evaluate the reliability of a rat tuberculous wound model established by injecting Bacillus Calmette-Guérin (BCG). Methods The experimental research was conducted. According to the random number table, fifteen 6-week-old male Sprague-Dawley rats were divided into normal control group and infection group, with 3 rats in normal control group and 12 rats in infection group. Rats in infection group were injected with Freund's complete adjuvant, 3 weeks later, they were injected subcutaneously with BCG bacterial solution to establish a model of tuberculous wounds in rats; rats in normal control group did not receive any treatment. On the 8th, 15th, 32nd, and 43rd day of infection, the skin condition at the injection sites of the rats in infection group was observed roughly. Skin tissue at the injection sites of 3 rats in infection group at each corresponding time point stated above and skin tissue at the corresponding sites of the rats in normal control group were stained with hematoxylin-eosin to observe the cell arrangement, necrosis and inflammation. On 43rd day of infection, acid-fast staining was performed on the skin tissue at the injection sites of the rats in infection group to observe the distribution of bacteria. Results On the 8th, 15th, 32nd, and 43rd day of infection, tuberculous wound lesions were gradually developed at the skin tissues at the injection sites of the rats in infection group. The cells of the diseased tissue of the rats in infection group arranged disorderly or concentrically, and the number of granulomas and necrotic cells gradually increased, while the skin tissue cells in the corresponding parts of the rats in normal control group arranged regularly with no inflammatory cell infiltration. On the 43rd day of infection, a large number of rod-shaped bacteria were observed in the skin tissue at the injection sites of the rats in infection group. Conclusions The rat tuberculous wound model established using BCG is stable and reliable, which can meet the experimental requirements.
- Models, animal /
- Tuberculosis, cutaneous /
- Rats /
- Bacillus Calmette-Guérin
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HTML全文

1 感染组大鼠感染不同时间点注射部位皮肤组织形成的病灶表现。1A.感染第8天，注射部位皮肤稍隆起（→）；1B.感染第15天，注射部位皮下脓肿较图1A明显（→）；1C.感染第32天，注射部位隆起的脓肿出现破溃；1D.感染第43天，脓肿破溃处无愈合倾向；1E~1H.感染第43天，创面脓性分泌物经挤压由感染灶内不断溢出的过程

下载: 全尺寸图片幻灯片

2 感染组大鼠感染不同时间点注射部位皮肤组织与正常对照组大鼠对应皮肤组织形态学观察苏木精-伊红×400。2A.正常对照组大鼠组织细胞排列规律；2B.感染组大鼠感染第8天,细胞分布杂乱、细胞大量聚集；2C.感染组大鼠感染第15天,可见肉芽肿形成（→）；2D.感染组大鼠感染第32天,肉芽肿数量增多（→）；2E.感染组大鼠感染第43天,细胞排列疏松，部分细胞坏死（→）

下载: 全尺寸图片幻灯片

3 感染组大鼠感染第43天注射部位皮肤组织抗酸染色结果。3A.细菌(红色)在组织内呈团簇样聚集石碳酸品红×600；3B.大量呈杆状的细菌菌体聚集在病灶中石碳酸品红×1 000

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参考文献(23)

[1]	World Health OrganizationGlobal tuberculosis report2020-05-31https://www.who.int/teams/global-tuberculosis-programme/data World Health Organization.Global tuberculosis report[R/OL].2018[2020-05-31]. https://www.who.int/teams/global-tuberculosis-programme/data.
[2]	World Health OrganizationGlobal tuberculosis report2020-05-31https://www.who.int/teams/global-tuberculosis-programme/data World Health Organization.Global tuberculosis report[R/OL].2017[2020-05-31]. https://www.who.int/teams/global-tuberculosis-programme/data.
[3]	贾赤宇.结核性创面——一个被忽视且值得重视的临床问题[J/CD].中华损伤与修复杂志:电子版,2014,9(4):9-11.DOI: 10.3877/cma.j.issn.1673-9450.2014.04.003.
[4]	SahinN,AydinNE,SenolM,et al.Longstanding skin ulcers due to Mycobacterium tuberculosis in a healthy man[J].Trop Biomed,2010,27(1):120-124.
[5]	荣美玉,贾赤宇.单中心结核性创面与非结核性慢性难愈性创面患者的特点[J].中华烧伤杂志,2019,35(2):90-94.DOI: 10.3760/cma.j.issn.1009-2587.2019.02.003.
[6]	常娜,贾赤宇,刘真,等.235例肺外结核性创面患者流行病学调查[J].中华烧伤杂志,2015,31(2):122-124.DOI: 10.3760/cma.j.issn.1009-2587.2015.02.011.
[7]	贾赤宇,李鹏程,程琳,等.外科干预治疗模式在窦道型结核性创面中的临床应用[J].中华烧伤杂志,2016,32(6):326-330.DOI: 10.3760/cma.j.issn.1009-2587.2016.06.003.
[8]	吕晓武,贾赤宇,冯胜娟,等.胸壁结核性创面外科治疗进展[J].感染、炎症、修复,2014,15(2):122-124.DOI: 10.3969/j.issn.1672-8521.2014.02.021.
[9]	秦云贺,王艺红,郭庆龙,等.结核分枝杆菌菌悬液光密度值与菌落计数的关联性[J].中国感染控制杂志,2016,15(3):150-154.DOI: 10.3969/j.issn.1671-9638.2016.03.002.
[10]	DannenbergAM.Liquefaction and cavity formation in pulmonary TB: a simple method in rabbit skin to test inhibitors[J].Tuberculosis (Edinb),2009,89(4):243-247.DOI: 10.1016/j.tube.2009.05.006.
[11]	HusainAA,DaginawalaHF,WarkeSR,et al.Investigation of immune biomarkers using subcutaneous model of M. tuberculosis infection in BALB/c mice: a preliminary report[J].Immune Netw,2015,15(2):83-90.DOI: 10.4110/in.2015.15.2.83.
[12]	张同威,汪毅平,贾赤宇.清创后外用碱性成纤维细胞生长因子对新西兰兔结核性创面愈合的影响[J].中华烧伤杂志,2019,35(2):95-103.DOI: 10.3760/cma.j.issn.1009-2587.2019.02.004.
[13]	汪毅平,刘真,张亚洁,等.结核性创面大鼠模型中巨噬细胞极化改变[J/CD].中华损伤与修复杂志:电子版,2018,13(1):30-36.DOI: 10.3877/cma.j.issn.1673-9450.2018.01.007.
[14]	ChenL,LiuZ,SuY,et al.Characterization of Mycobacterium marinum infections in zebrafish wounds and sinus tracts[J].Wound Repair Regen,2017,25(3):536-540.DOI: 10.1111/wrr.12540.
[15]	ImamuraT,IyamaK,TakeyaM,et al.Role of macrophage tissue factor in the development of the delayed hypersensitivity reaction in monkey skin[J].Cell Immunol,1993,152(2):614-622.DOI: 10.1006/cimm.1993.1317.
[16]	汪毅平,张同威,贾赤宇.结核性创面动物模型的研究进展[J/CD].中华损伤与修复杂志:电子版,2017,12(4):309-311.DOI: 10.3877/cma.j.issn.1673-9450.2017.04.014.
[17]	ZhangG,ZhuB,ShiW,et al.Evaluation of mycobacterial virulence using rabbit skin liquefaction model[J].Virulence,2010,1(3):156-163.DOI: 10.4161/viru.1.3.11748.
[18]	DongX,LuoY,GaoQ,et al.Effects of MBL-associated serine protease-2 (MASP-2) on liquefaction and ulceration in rabbit skin model of tuberculosis[J].Microb Pathog,2016,99:282-286.DOI: 10.1016/j.micpath.2016.08.037.
[19]	王明珠,史大中,杨爱军,等.分枝杆菌所致家兔皮肤液化病理模型研究[J].微生物与感染,2008,3(4):208-211,218.DOI: 10.3969/j.issn.1673-6184.2008.04.004.
[20]	SugaM,DannenbergAM,HiguchiS.Macrophage functional heterogeneity in vivo. Macrolocal and microlocal macrophage activation, identified by double-staining tissue sections of BCG granulomas for pairs of enzymes[J].Am J Pathol,1980,99(2):305-323.
[21]	HiguchiS,SugaM,DannenbergAM,et al.Persistence of protein, carbohydrate and was components of tubercle bacilli in dermal BCG lesions[J].Am Rev Respir Dis,1981,123(4 Pt 1):397-401.DOI: 10.1164/arrd.1981.123.4.397.
[22]	ChenH,LiuX,MaX,et al.A new rabbit-skin model to evaluate protective efficacy of tuberculosis vaccines[J].Front Microbiol,2017,8:842.DOI: 10.3389/fmicb.2017.00842.
[23]	GutierrezMG,MasterSS,SinghSB,et al.Autophagy is a defense mechanism inhibiting BCG and Mycobacterium tuberculosis survival in infected macrophages[J].Cell,2004,119(6):753-766.DOI: 10.1016/j.cell.2004.11.038.