中华皮肤科杂志 ›› 2010, Vol. 43 ›› Issue (4): 230-234.

• 论著 • 上一篇    下一篇

单核细胞趋化蛋白-1 DNA疫苗治疗姥鲛烷诱导BALB/c狼疮鼠模型的实验研究

王贵红1,李向培2,厉小梅3,汪国生4,张宏胡闻5,王怡平5   

  1. 1. 安徽医科大学附属省立医院风湿免疫科
    2. 安徽医科大学附属省立医院
    3. 合肥市安徽省立医院风湿科
    4. 安徽省立医院风湿免疫科
    5.
  • 收稿日期:2009-07-28 修回日期:2010-01-12 出版日期:2010-04-15 发布日期:2010-04-07
  • 通讯作者: 王贵红 E-mail:wgh-70@sohu.com

Experimental study on pristane-induced BALB/c mouse models for lupus immunized with monocyte chemoattractant protein-1 （MCP-1） DNA vaccine

  • Received:2009-07-28 Revised:2010-01-12 Online:2010-04-15 Published:2010-04-07
  • Contact: Guihong Wang E-mail:wgh-70@sohu.com

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摘要：

目的 应用单核细胞趋化蛋白-1（MCP-1）基因片段的DNA疫苗免疫姥鲛烷诱导的BALB/c狼疮鼠模型，研究MCP-1 DNA疫苗对狼疮鼠模型的免疫抑制作用以及对肾脏的保护效果。方法 BALB/c小鼠随机分为4组：狼疮鼠模型组、疫苗组、空质粒组、正常对照组。制备编码MCP-1基因片段的DNA疫苗，接种于疫苗组。测定各组小鼠体质量、24 h尿蛋白定量，ELISA法测定血清MCP-1抗原和抗体水平，免疫荧光法检测血清ANA；HE染色光镜下观察小鼠肾脏病理变化，免疫组化检测MCP-1、CD68在肾脏内的表达。结果 24周时狼疮鼠模型组、疫苗组、空质粒组和正常对照组间体质量和24 h尿蛋白定量差异有统计学意义，F值分别为20.31，6.74，P值均 < 0.05，LSD检验示疫苗组与狼疮鼠模型组和空质粒组之间差异有统计学意义（P值均 < 0.01）。狼疮鼠模型组、疫苗组、空质粒组和正常对照组血清MCP-1抗原水平分别为（572.6 ± 58.55）、（169.52 ± 28.71）、（601.98 ± 83.43）和（61.11 ± 66.12） ng/L，差异有统计学意义（F = 143.09，P < 0.05），LSD检验示疫苗组与狼疮鼠模型组、空质粒组之间差异有统计学意义（P < 0.01）；四组的血清MCP-1抗体水平分别为（357.88 ± 82.41）、（770.14 ± 220.91）、（294.25 ± 177.22）和（129.73 ± 168.24） ng/L，方差分析示F = 15.81，P < 0.01，LSD检验示疫苗组与狼疮鼠模型组、空质粒组和正常对照组之间差异有统计学意义。ANA抗体阳性率四组间差异无统计学意义。疫苗组和正常对照组与狼疮鼠模型组和空质粒组比较，肾脏损伤明显减轻，肾脏内MCP-1和CD68阳性细胞明显减少。结论 MCP-1质粒DNA疫苗能够有效抑制狼疮鼠模型血清中MCP-1抗原表达，并诱导MCP-1抗体水平升高。

关键词: 关键词：红斑狼疮, 系统性, DNA疫苗, MCP-1

Abstract:

Objective To immunize pristane-induced BALB/c mouse models for lupus with MCP-1 DNA vaccine, and to study the immunosuppressive effect and protective effect on kidney of the DNA vaccine in the murine lupus model. Methods BALB/c mice were randomly divided into four groups. Pristane-induced lupus mice were immunized with MCP-1 DNA vaccine （MCP-1 vaccine group）, empty plasmids （empty plasmid control group） or remained non-vaccinated (lupus model group). Normal mice receiving neither induction nor vaccination served as the normal control. Subsequently, mice were monitored for body weight and 24-hour protein excretion; ELISA was used to measure the serum levels of MCP-1antibody and antigen, and immunofluorescence method to detect serum ANA. Hematoxylin and eosin staining was performed to observe pathological changes of murine kidney, immunohistochemistry to assess the expression of CD68 and MCP-1 in renal tissue. Results At week 24, there was a significant difference in body weight and 24-hour protein excretion among the 4 groups （F = 20.31, 6.74, both P < 0.05）, and LSD test showed that MCP-1 vaccine group differed significantly from the model group and empty plasmid group in the two parameters （both P < 0.01）. The serum levels at week 24 were （572.6 ± 58.55） ng/L, （169.52 ± 28.71） ng/L, （601.98 ± 83.43） ng/L and（61.11 ± 66.12） ng/L for MCP-1 antigen （F = 143.09, P < 0.05）, （357.88 ± 82.41） ng/L, （770.14 ± 220.91） ng/L, （294.25 ± 177.22） ng/L and （129.73 ± 168.24） ng/L for MCP-1 antibody （F = 15.81, P < 0.01） in lupus model group, MCP-1 vaccine group, empty plasmid group and normal control group, respectively; LSD test revealed that the MCP-1 vaccine group was significantly different from the lupus model group and empty plasmid group in the serum level of MCP-1 antigen and from the other 3 groups in the level of MCP-1 antibody （all P < 0.01）. No significant difference was observed in the positivity rate of antinuclear antibody among the 4 groups. In MCP-1 vaccine group and normal control group, renal damage was much milder together with a reduction in the number of MCP-1- and CD68-positive cells in renal tissue in comparison with the other two groups. Conclusion MCP-1 DNA vaccines can efficiently inhibit the expression of MCP-1 antigen and induce the production of MCP-1 antibody in mouse model for lupus.

Key words: Key words: lupus erythematosus, systemic, DNA vaccine, MCP-1