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目的建立活性染色质诱导系统性红斑狼疮(SLE)样小鼠模型。方法从ConA活化的BALB/c小鼠脾淋巴细胞中提取活性染色质,分别于d 0、d 14、d 21和d 28以染色质100μg在BALB/c小鼠尾根部及背部皮内注射免疫4次,诱导SLE样小鼠模型。目测半定量尿蛋白试纸法检测动物的尿蛋白变化,HE染色法检查动物的肾脏、脾脏病理改变,计算动物的胸腺和脾脏指数,MTT法检测ConA和LPS诱导的T、B淋巴细胞增殖反应,全自动生化分析仪检测血清中Crea和BUN水平,ELISA法检测小鼠血清中ANA、抗dsDNA、IgG1、IgG2a、IL-10、IFN-γ水平,流式细胞术检测脾脏T、B淋巴细胞亚群变化。结果诱导模型小鼠尿蛋白水平升高,出现肾小球肾炎、脾脏增生等病理改变;脾脏指数明显升高,LPS诱导的B淋巴细胞增殖反应增强;血清Crea、BUN、ANA、抗dsDNA、IgG1、IgG2a、IL-10和IFN-γ水平明显升高;脾脏CD19+、CD19+CD21+、CD19+CD23+、CD19+IgD+B淋巴细胞亚群百分比明显升高,CD4+CD25+T淋巴细胞百分比明显下降。结论 ConA活化淋巴细胞的染色质免疫同系BALB/c小鼠成功诱导了SLE样小鼠模型,其血清学、组织病理学及免疫学方面特征与人类SLE临床特征表现相似。
Objective To establish a reactive chromatin-induced systemic lupus erythematosus (SLE) -like mouse model. Methods The active chromatin was extracted from splenic lymphocytes of ConA-activated BALB / c mice and immunized intradermally at the tail root and the back of BALB / c mice with 100 μg of chromatin at d 0, d 14, d 21 and d 28, respectively 4 times, induced SLE-like mouse model. The proteinuria of urinary protein was detected by semi-quantitative urine protein test paper method. The pathological changes of kidney and spleen were examined by HE staining. Thymus and spleen indexes of animals were calculated. The proliferation responses of T and B lymphocytes induced by ConA and LPS were detected by MTT assay. Serum levels of Crea and BUN were detected by automatic biochemical analyzer. Levels of ANA, anti-dsDNA, IgG1, IgG2a, IL-10 and IFN-γ in sera of mice were detected by ELISA. T and B lymphocyte subsets in spleen were detected by flow cytometry Group changes. Results The level of urinary protein in induced model mice increased, pathological changes such as glomerulonephritis and spleen hyperplasia were observed. The index of spleen was obviously increased and the proliferation of B lymphocytes was enhanced by LPS. Serum Crea, BUN, ANA, anti-dsDNA, IgG1 , The levels of IgG2a, IL-10 and IFN-γ were significantly increased. The percentages of CD19 +, CD19 + CD21 +, CD19 + CD23 +, CD19 + IgD + B lymphocyte subsets in spleen were significantly increased and the percentage of CD4 + CD25 + T lymphocytes was significantly decreased . CONCLUSIONS: ConA-activated lymphocyte chimeric BALB / c mice successfully induced a SLE-like mouse model with similar serological, histopathological and immunological characteristics as the clinical features of human SLE.