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作 者:梁海梅[1] 于化鹏[1] 夏虎[1] 易丽[1] 方泽葵[1] 李忠丽[1]
机构地区:[1]南方医科大学珠江医院呼吸内科,广东广州510282
出 处:《热带医学杂志》2013年第5期570-572,594,F0003,共5页Journal of Tropical Medicine
摘 要:目的研究NLRP3、IL-33在脂多糖(LPS)诱导的RAW264.7巨噬细胞炎症模型中的表达及格列本脲对其表达的影响。方法培养RAW264.7巨噬细胞,将其分为空白对照(A)组、NLRP3抑制剂格列本脲预处理+LPS(B)组,LPS模型(C)组;B、C组采用LPS刺激RAW264.7巨噬细胞建立炎症模型,B组在LPS刺激前30min给予格列本脲处理,空白对照组只给予培养基培养。采用蛋白免疫印迹法(Western-blot)检测细胞中NLRP3蛋白的表达;酶联免疫吸附试验(ELISA)检测细胞培养上清中1L-33的含量;四氮唑盐(MTT)法测定各组细胞的增殖能力。结果 C组中NLRP3蛋白表达均高于A组、B组(均P<0.05),B组NLRP3蛋白表达高于A组(P<0.05)。C组IL-33含量比A、B组高(P<0.05),A组和B组之间差异无统计学意义。MTT试验中C组细胞增殖率高于A组、B组(P<0.05),而A、B组差异无统计学意义。结论 NLRP3、IL-33在LPS诱导RAW264.7的巨噬细胞中表达升高,而格列本脲干预后可抑制NLRP3的表达及IL-33的分泌。Objective To test the expression of NLRP3 and IL-33 in lipopolysaccharides (LPS)-induced RAW264,7 cells before and after glyburide intervention. Methods RAW264.7 cells were divided into three groups: normal control group (group A), NLRP3 inhibitor(glyburide) +LPS group (group B) and LPS-induced cell inflammation group (group C). In groups B and C, LPS was used for building the RAW264.7 cell inflammation model. Glyburide was added 30 minutes before LPS exposure in group B. NLRP3 expression was identified by Western-blot method and IL-33 production was detected by ELISA in cell supernatant. MTF method was used to detect the cell proliferation. Results The protein expression level of NLRP3 in group C was significantly higher than those in groups A and B (P〈0.05). NLRP3 expression level in group B was higher than that in group A (P〈0.05). And the production of IL-33 in group C was significantly higher than those in groups A and B (P〈0.05), but no significant difference was observed between groups A and B. The proliferation state in group C was higher than those in groups A and B (P〈0.05), but no significant difference was noted between groups A and B. Conclusion LPS-induced RAW264.7 cells promote NLRP3 and IL-33 expression, and glyburide can inhibit the expression of NLRP3 and IL-33.
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