摘要
目的探讨C5a在阿尔茨海默病(AD)小胶质细胞病变模型中释放炎性因子的作用及其机制,明确C5a受体拮抗剂对上述病变过程的干预作用。方法制备Aβ1-42寡聚体,用Aβ1-42寡聚体刺激BV-2小胶质细胞建立AD小胶质细胞病变模型。实验组分为Aβ1-42干预组、Aβ1-42+C5a组、Aβ1-42+C5aRA组、Aβ1-42+C5a+C5aRA组,分别加入不同浓度的C5a、C5aRA、C5a+C5aRA进行干预,以小胶质细胞无干预组为空白对照。通过流式细胞术测定细胞表面C5a受体(CD88)的表达、ELISA法检测上清液中炎症因子TNF-α的浓度。结果 Aβ1-42刺激小胶质细胞分泌TNF-α,Aβ1-42组TNF-α浓度与空白对照组相比明显增加(P<0.05),该组CD88表达高于空白对照组(P<0.01);Aβ1-42+C5a组的TNF-α浓度高于单纯Aβ1-42组(P<0.05),Aβ1-42+C5aRA组TNF-α浓度低于单纯Aβ1-42组(P<0.01),而Aβ1-42+C5a组及Aβ1-42+C5aRA组的CD88表达与单纯Aβ1-42组相比无显著差异(P>0.05)。结论 C5a能刺激小胶质细胞分泌炎症因子TNF-α,且与Aβ1-42有协同作用,而C5aRA则能有效抑制该通路。
Objective To investigate the role of CSa in the release of TNF-a from microglial ceils and its related mechanism, and identify the intervention effect of CSaR antagonist （CSaRA） in the pathological change of Alzheimer＇s disease （AD）. Methods Soluble Aβ1-42 oligomer was prepared, identified by atomic force microscopy. Then Aβ1-42 oligomer alone, or in a combination with different levels of CSa, CSaRA, or CSa ＋ CSaRA was used to treat cultured BV2 cells. The non-treated BV2 cells served as controls. ELISA was used to detect the concentration of TNF-α, and flow cytometry to analyze the expression of CSa receptor （CD88）. Results Aβ1-42 oligomer significantly stimulated BV2 cells to release TNF-α and CD88, compared to the control group （ P 〈 0.05 and P 〈 0.01, respectively）. CSa further increased the release of TNF-α （ P 〈 0.05 vs Aβ1-42 group）. In contrast, CSaRA inhibited remarkably the release of TNF-α stimulated by Al31-42 （P〈0.01）. Never- theless, CSa or CSaRA didn＇t show a significant effect on the expression of CD88 in BV2 cells that was elevated by Aβ1-42 treatment （P〉0.05）. Conclusion CSa promotes the release of TNF-α from microglial cells; Aβ1-42 can further boost the release and CSaRA can inhibite TNF-a secretion of microglial cells significantly.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2013年第3期265-268,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
广州市科技计划项目基金(2010Y1-C101)
关键词
阿尔茨海默病
补体
Β-淀粉样蛋白
Alzheimer＇s disease
complement system
amyloid β protein
