TLR4基因在CD38--小鼠B细胞发育障碍中的作用-药物分析专业论文.docxVIP

PAGE PAGE VI 摘要 (Immunohistochemistry, IHC-P)观察 CD38-/-和 CD38-/-TLR4mut 小鼠脾脏的组 织学差异； 7. 通过流式细胞术（Flow Cytometry）和相关表面抗体（包括 CD21 FITC、CD24 PE、 CD45R PE Cy5.5 和 IgD APC 等）染色检测 B 细胞的细胞发育、细胞 周期和细胞凋亡情况。 结果： 1. LPS 刺激可以显著促进小鼠 B 细胞的发育；而与 WT 小鼠相比，CD38-/-小鼠 的 B 细胞的发育成熟度明显下降；CD38-/-小鼠的 B 细胞周期被阻滞在 S 期； 2. 成功制备并鉴定了 CD38-/-TLR4mut 小鼠； 3. 与 CD38-/-小鼠相比较，CD38-/-TLR4mut 小鼠脾脏 B 细胞中 TLR4 基因和炎性 因子 MCP-1 和 TNF-α的 mRNA 表达水平显著下降； 4. 与 CD38-/-小鼠相比较，CD38-/-TLR4mut 小鼠脾脏 B 细胞中 TLR4 蛋白表达下 降，而 Sirt1、NF-κB（p65）和 AC-NF-κB 蛋白表达水平未见明显改变； 5. 与 CD38-/-小鼠相比较，CD38-/-TLR4mut 小鼠脾脏巨噬细胞和中性粒细胞数目 减少； 6. 与 CD38-/-小鼠相比较，CD38-/-TLR4mut 小鼠脾脏 B 细胞发育阻滞更加严重， 集中表现在 T1 向 T2 的转型障碍；但细胞周期和细胞凋亡未见明显影响。 结论： 1. TLR4 基因参与了 CD38 基因敲除所引起的 B 细胞的发育障碍，且与 Sirt1 途 径无相关性； 2. CD38 基因的敲除可通过抑制 TLR4 基因的表达，降低其下游炎性因子 MCP-1 和 TNF-α的表达，减少炎性细胞如巨噬细胞和中性粒细胞的浸润； 3. CD38-/-和 TLR4mut 协同抑制 B 细胞的发育成熟，特别是 T1 向 T2 的转型。 关键词： Toll 样受体 4 ；CD38；B 细胞；发育 Abs Abstract ABSTRACT Objective： TLR4 is an important molecule related to innate immune, it play an important role in the bodys inflammatory response by recognizing pathogen associatedmolecular patterns. At the meantime, TLR4 is also involved in the development and progression of cancer, atherosclerosis and autoimmune diseases. Studies showed that CD38 gene knockout mice have the obstacled B cell development,especially obstacled at immature B cells phase in which the apoptosis of T1 cells is decreasing and the development of T2 cells is obstacled. Our previously study found that the expression of TLR4 gene is specifically expressed in mature B cells in bone marrow, and it’s expression is increased significantly with the migration of mature B cells from bone marrow to spleen. The B cell development of TLR4mut mice is also obstacled. In addition, the expression of TLR4 in spleen is also decreased in CD38-/- mice. These results hint that the influence of TLR4 and CD38 on B cell development may have some inner relations.Our study aims at, to further explore the role of CD38 and TLR4 in B cell developmen