兰州大学机构库 >数学与统计学院
一氧化氮对巨噬细胞亚型分化影响的实验研究
Alternative TitleThe impact of nitric oxide on macrophage subtype differentiation experiments
胡旭堂
Thesis Advisor王志禄
2014-05-28
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
Keyword动脉粥样硬化 RAW264.7巨噬细胞 SNAP M1、M2亚型
Abstract背景 动脉粥样硬化是一种发生在血管壁的慢性炎症过程,巨噬细胞在其中发挥着关键作用。不同亚型巨噬细胞对粥样斑块内脂质核的形成及板块的稳定性有不同的影响,因此,有人提出通过调控粥样斑块内巨噬细胞亚型的分化来改变动脉粥样硬化病程的进展。目的 本实验通过观察本实验通过观察S-亚硝基-N-乙酰-DL-青霉胺(SNAP)对巨噬细胞亚型分化标志物CD86、iNOS 、Arg-I、MR 及内质网应激标志物p-PERK、CHOP表达的影响,旨在探讨○1NO对巨噬细胞亚型分化的影响及其机制,○2NO的免疫调节功能在动脉粥样硬化病程中的影响。方法 以RAW 264.7巨噬细胞为研究对象,分为空白对照组和不同浓度SNAP干预组,应用RT-PCR法检测RAW264.7巨噬细胞亚型分化标志物M1(iNOS,CD86)、M2(Arg-I,MR)及CHOP mRNA的表达,应用Western blot技术检测iNOS及ERS通路中相关蛋白CHOP、P-PERK的表达。结果 与空白对照组比较,SNAP干预组iNOS、CD86、CHOP mRNA的表达均明显降低(P﹤0.05),Arg-I mRNA表达明显升高(P﹤0.05),而MR mRNA表达尽管升高,但无统计学意义(P﹥0.05);SNAP组CHOP、iNOS、p-PERK蛋白表达均明显低于对照组(P﹤0.05)。结论 NO可以抑制巨噬细胞向M1亚型分化,NO可抑制巨噬细胞内质网应激,两者可能相关。
Other AbstractBackground. Atherosclerosis is a chronic inflammatory process occured in the vessel wall, macrophages play a key role during this process. The different subtype of macrophages influence the disease process in different way, so atherosclerosis treatment using medicines in order to control subsets macrophages differentiation might be a new therapeutic aim. Object. To investigate the effect and mechanism of NO to macrophage phenotype differentiation in atherosclerosis by observing the changes of CD86, iNOS, Arg-I, MR and p-PERK and CHOP after stimulated by SANP. Methods. The RAW 264.7 macrophages were seeded in six wells (106/ ml in each) for 24 h before intervention of SNAP in different concentration(30, 100, 300, 400 and 500 µmol/L). Total RNA of cells was extracted after intervention for 24 hours. Detecting the phenotype marker iNOS, CD86 (as M1 phenotypes markers), MR, Arginase-I (Arg-I) (as M2 phenotypes markers) respectively by real time PCR on RNA level to determine whether SNAP had influenced in macrophage subtypes differentiation. Detected CHOP, p-PERK the response proteins of ERS by Western blotting on protein level to determine if this change is related with ERS. Results. iNOS and CD86 of RAW 264.7 cell decreased remarkably(P﹤0.05) compared with control group after stimulation by different concentration of SNAP for 24h as well as CHOP and p-PERK, in contrast Arg-I increased remarkably(P﹤0.05), however there were no changes to MR of RAW 264.7 cell; last the p-PERK, iNOS and CHOP protein were decreased remarkably after stimulated by different concentration of SNAP, this changes correspond with the concentration of SNAP . Conclusion. SNAP can suppress macrophage differentiation to M1 subtype and it is related with endoplasmic reticulum stress (ERS).
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Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/224421
Collection数学与统计学院
Recommended Citation
GB/T 7714
胡旭堂. 一氧化氮对巨噬细胞亚型分化影响的实验研究[D]. 兰州. 兰州大学,2014.
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