摘要
为研究蛋白酶体激活因子REGγ对脂多糖(lipopolysaccharides,LPS)介导的腹膜炎的调节作用,采用腹腔注射LPS的方法以建立小鼠腹膜炎模型.首先,通过骨髓细胞过继转移的方法构建骨髓嵌合体小鼠;利用存活率实验检测REGγ敲除对小鼠生存率的影响;然后,通过酶联免疫吸附测定实验检测促炎因子的表达.结果显示,与野生型(WT)小鼠相比,REGγ敲除(KO)小鼠表现为较低的存活率,且细胞因子TNFα(tumor necrosis factor-α)、IL-1β(interleukin-1β)和MCP-1(monocyte chemoattractant protein-1)的表达也较低.提取WT和KO小鼠来源的骨髓细胞,分别过继转移至X-射线辐照过的WT小鼠,获得WTchimera和KO^(chimera)骨髓嵌合体小鼠.腹腔注射LPS于WTchimera和KO^(chimera)嵌合体小鼠,结果发现,KO^(chimera)也表现为较低的存活率和TNFα、IL-1β和MCP-1表达.研究结果表明,REGγ在造血系统来源的炎症细胞中敲除,可以加速LPS介导的小鼠存活率下降,且抑制机体促炎因子TNFα、IL-1β和MCP-1的表达.
Mouse peritonitis model is established by LPS peritoneal injection to analyze the roles of proteasome activator REGγ in LPS-mediated peritonitis. Bone marrow chimera mice are generated by adoptive transfer of bone marrow cells. Survival assay is used to research the effect of REGγ deficiency on survival rate. Expression of proinflammatory factors are detected via enzyme-linked immunosorbent assay（ ELISA）. As compared to the wild-type（ WT） mice,the REGγ-knockout（ KO） mice shows significantly lower survival rate and lower expression of the proinflammatory factors TNFα,IL-1β and MCP-1. Furthermore,bone marrow from WT and KO mice is isolated and transferred adoptively to X-ray irradiated WT mice to generate WTchimeraand KO^（chimera）bone marrow chimera mice. The results suggest that KO^（chimera）mice also manifest obviously lower survival rate and proinflammatory factors expression of TNFα,IL-1β and MCP-1 after peritoneal injection of LPS to WTchimeraand KO^（chimera）. Thus,these data suggest that REGγ deficiency in inflammatory cells of hematopoietic origin accelerates LPS-mediated decline of survival rate and suppresses expression of proinflammatory factors TNFα,IL-1β and MCP-1.
出处
《深圳大学学报（理工版）》
EI
CAS
CSCD
北大核心
2017年第5期471-475,共5页
Journal of Shenzhen University(Science and Engineering)
基金
国家自然科学基金资助项目(31401217)
中国博士后科学基金资助项目(2014M0560672)~~
关键词
腹膜炎
蛋白酶体
过继转移
骨髓嵌合体
造血系统
酶联免疫吸附测定法
peritonitis
proteasome
adoptive transfer
bone marrow chimera
hematopoietic system
enzyme-linkedimmunosorbent assay
