C57BL／6小鼠结节病样肉芽肿模型的建立及鉴定  

Establishment and identification of a C57B/6 mouse sarcoidosis granuloma model

作　　者：黄燕[1] 路聪哲[1] 王彩彩[1] 姜毅[1] 王晓阳[1] 段蕴铀[1] 

机构地区：[1]海军总医院呼吸内科全军结节病中心,北京100048

出　　处：《中华结核和呼吸杂志》2013年第8期587-591,共5页Chinese Journal of Tuberculosis and Respiratory Diseases

基　　金：中国博士后科学基金面上基金（2012M521888）

摘　　要：目的建立并鉴定分枝杆菌过氧化物歧化酶多肽（SodA）诱导的C57BL／6小鼠结节病样肉芽肿模型。方法将30只雌性C57BL／6小鼠按数字随机表法分为5组：复合组（SodA＋脂糖）、SodA组，不完全弗氏佐剂（IFA）组、琼脂糖组及空白对照组，每组6只。第1天，复合组、SodA组给予50μgSodA与0．25mlIFA，混合后背部皮下注射；IFA组、琼脂糖组分别给予IFA0．25ml及PBS0．25ml背部皮下注射；空白对照组未予任何处理。第14天，复合组给予50μg SodA与6000粒琼脂糖4B珠（溶于0．5mlPBS）共价偶联后尾静脉注射；Soda组给予50μgSoda（溶于PBS0．5m1）尾静脉注射；IFA组、琼脂糖组给予6000粒琼脂糖4B珠（溶于0．5mlPBS）尾静脉注射；空白对照组未予任何处理。第22天，解剖小鼠，观察淋巴结、肺部大体标本及组织学改变；免疫组织化学检测肉芽肿内巨噬细胞和CD4^＋细胞；检测BALF中细胞分类；流式细胞术检测BALF中CD4＋／CD8^＋及肺组织内干扰素．1与IL-12水平。结果复合组肺部与淋巴结均可见结节病样肉芽肿，SodA组淋巴结可见结节病样肉芽肿，但肺部未见肉芽肿，IFA组、琼脂糖组及空白对照组淋巴结及肺部均未见结节病样肉芽肿；肉芽肿中心巨噬细胞特异性抗原Mac-2表达阳性，周围CD4^＋细胞表达阳性；复合组、SodA组BALF中淋巴细胞百分比（分别为19．4％±6．5％、22．3％±8．5％）与IFA组、琼脂糖组及空白对照组（分别为8．5％±4．3％、7．7％±3．4％、0．8％±0．6％）相比均明显增高（均P〈0．05）；复合组、SodA组BALF中CD4^＋／CD8^＋（分别为3．5±1．4、3．2±1．1）与IFA组、琼脂糖组（分别为1．2±0．5、1．0±0．4）相比均明显增高（均P〈0．05）；复合组、SodA组肺组织内干扰素-1、IL-12水平[干扰素-γ：（32．9±9．7）ng／L、（26．4±7．2）ng／L；IL-12：（29．6±9．4）ng／L、（26．1±8Objective To establish a C57BL/6 mouse sarcoidosis granuloma model elicited by mycobacterial superoxide dismutase A peptide （SodA）. Methods Thirty female C57BL/6 mice were randomly divided equally into 5 groups ： a combination （ Soda ＋ Sepharose） group, a Soda group, a IFA （incomplete Freund＇ s adjuvant）group, a sepharose group and a blank control group. On the first day, the combination group and the Soda group were sensitized by subcutaneous injection of 50 μg Soda incorporated into IFA 0. 25 ml. The IFA group and the Sepharose group were treated with subcutaneous injection of IFA 0. 25 ml and PBS 0. 25 ml respectively, while the blank control group was not given any treatment. On the 14th day, the combination group was challenged by tail vein injection of 50 μg SodA covalently coupled to 6000 agarose 4B beads （ in PBS 0. 5 ml）. The SodA group was challenged by tail-vein injection of 50μg SodA （ in PBS 0. 5 ml）. The IFA group and the Sepharose group were treated by tail-vein injection of 6000 agarose 4B beads（ in PBS 0. 5 ml） , while the blank control group was not given any treatment. On the 22th day, the mice were dissected and the gross and pathological changes of lymph nodes and lungs were observed, hnmunohistochemisty was used to identify Mac-2 and CD4^±T in granuloma. Counts and differentials of BALF cells were measured. CD4^4/CD8^± in BALF and cytokines （IFN-γ/and IL-12 ） levels in the lungs were detected by flow cytometry. Results Enlargement of peripheral and pulmonary hilar lymph nodes were found in the combination group and the Soda group, and sarcoidosis granuloma was found in the lymph nodes and lungs of the combination group. Sarcoidosis granuloma was also found in the lymph nodes but not in the lungs of the Soda group. No sarcoidosis granuloma was observed in the lungs and lymph nodes of the IFA group, the Sepharose group and the blank control group. Macrophage specific antigen Mac-2 and CD4^± T were positive in the core and rim of the granuloma respective

关 键 词：结节病 肉芽肿 浆细胞 肺 模型 动物 

分 类 号：R-332[医药卫生]