系统性红斑狼疮(2)

Abstract

Abstract

Purpose:

Systemic lupus erythematosus (SLE) is a complex antoimmune disease characterized by the production of anti-double-stranded DNA (anti-dsDNA) antibodies, which can form immune complexes and deposit in tissues, causing inflammation and organ damage. SLE can affect multiple organs, including nervous system, digestive system, respiratory system, blood system, mucocutaneous, musculoskeletal, livers, kidneys, etc., mainly kidneys. The cause of the disease includes both genetic and environment factors. Because the pathogenesis of SLE is complex, there is no effective way to cure SLE.The immune system dysregulation in SLE involves both adaptive and innate immunity. Interleukin-6 (Interleukin-6, IL-6) is a proinflammatory cytokine, which plays an important role in the inflammatory response. Previous studies in a variety of autoimmune diseases have reported elevated serum IL-6 levels. Here, we Constructed a mouse model of SLE, to study the role of IL-6 in the SLE pathogenesis. Methods:

(1) Spleen lymphocytes from C57 Wt mice were stimulated by ConA, activated lymphocyte derived DNA (ALD-DNA) was extracted. Immunized C57 Wt mice with ALD-DNA and detected SLE associate indicators, including the anti-dsDNA antibodies, proteinuria, renal biopsy.

(2) Immunized IL-6KO mice as described above and detected SLE associate indicators.

(3) In vivo, measured the anti-double-stranded DNA antibodies, IL-6, IL-17 and TNF-a in immunized mice with ELISA . In vitro, stimulated Spleen lymphocytes from C57 Wt mice with ALD-DNA and measured the anti-dsDNA antibodies, IL-6, IL-17 and TNF-a in supernatants

(4) The Spleen and lymph nodes lymphocytes from immunized mice were isolated,

III

Abstract

the percentages of regulatory T cells (Treg) and activation of CD4+T cells were analyzed by flow cytometry. Spleen lymphocytes from immunited mice were cultured under Treg and Th1 differentiation condition, and the percentages of Treg and Th1 were analyzed by flow cytometry.

(5) Bone marrow cells from normal C57 mice were cultured under bone marrow-derived dendritic cells (BMDC) differentiation condition, stimulated BMDC with ALD-DNA, measured the IL-6 and TNF-a in supernatants. Spleen lymphocytes from normal C57 mice were cultured under Treg differentiation condition with the supernatants, the percentages of Treg were analyzed by flow cytometry. Blocking IL-6 or TNF-a with Anti-IL-6 or anti- TNF-a antibodies, the percentages of Treg were analyzed by flow cytometry. ?? Results:

(1) ALD-DNA can induce SLE in C57 Wt mice, with the production of anti-dsDNA antibodies, proteinuria, renal pathological damage.

(2) IL-6KO mice were resistant to the development of SLE, which suggested that IL-6 is crucial for SLE.

(3) ALD-DNA immunized C57 Wt mice produced high level of anti-dsDNA Abs, IL-6 in serum and in vitro ALD-DNA can induce the production of anti-dsDNA Abs, IL-6 , IL-17and TNF-a.

(4) ALD-DNA immunized C57 Wt mice had higher activation of CD4+ T cells and lower expression of Foxp3+ than IL-6KO immunized mice and control mice. (5) ALD-DNA stimulated DC could produced high levels of IL-6 and TNF-a. ALD-DNA stimulated DC could suppress FoxP3 expression via IL-6. Conclusion:

IL-6 secreated by BMDC can promote the progression of SLE by suppressing Treg. The knowledge on IL-6 not only fosters our understanding of the disease, but also provides insights in devising biomarkers and targeted therapies.

Key Words: SLE、 IL-6、Treg、BMDC

IV

目录

目录

摘 要 ........................................................................................................ I Abstract .................................................................................................. III 目录 ........................................................................................................ V 第一章

前言 ....................................................................................... 1

第一节 系统性红斑狼疮 ....................................................................................1

1.1.1 系统性红斑狼疮的病理表现 ............................................................................. 1 1.1.2 系统性红斑狼疮的致病原因 ............................................................................. 4 1.1.3 系统性红斑狼疮的治疗 ..................................................................................... 6

第二节 系统性红斑狼疮的研究现状 ................................................................8

1.2.1 IL-6与系统性红斑狼疮 ...................................................................................... 9 1.2.2 Treg与系统性红斑狼疮 ................................................................................... 11 1.2.3 DC与系统性红斑狼疮 ...................................................................................... 12 1.2.4 CD4 +T细胞与系统性红斑狼疮（图1.9） ..................................................... 17

第三节 本实验研究 ..........................................................................................18

1.3.1 研究目的和意义 ............................................................................................... 18 1.3.2 研究方法和内容 ............................................................................................... 19

第二章 实验材料和方法 .................................................................. 20

第一节 实验材料 ..............................................................................................20

2.1.1 实验小鼠 ........................................................................................................... 20 2.1.2 实验试剂 ....................................................................................... …… 此处隐藏：8407字，全部文档内容请下载后查看。喜欢就下载吧 ……