二甲双胍增强LPS激活的腹腔巨噬细胞在ATP刺激下的炎症小体活化

发布时间：2018-05-28 19:05

本文选题：二甲双胍 + 细胞焦亡　；参考：《中国药理学通报》2017年04期

【摘要】：目的以脂多糖(LPS)激活的腹腔巨噬细胞为炎症细胞模型,探讨糖尿病常用药物二甲双胍对胞外ATP诱导炎症小体活化并释放白细胞介素-1β(IL~(-1)β)的影响。方法C57BL/6小鼠腹腔注射30 g·L~(-1)巯基乙酸盐(thioglycollate,TG)诱导腹腔巨噬细胞大量产生;利用LPS+ATP处理激活炎症小体,采用碘化丙锭(PI)染色法检测二甲双胍对LPS+ATP诱导的腹腔巨噬细胞焦亡的影响,免疫印迹法检测该细胞胞内及上清中IL~(-1)β和caspase-1的表达水平;免疫荧光技术检测巨噬细胞P_2X_7嘌呤能ATP受体7(P_2X_7R)的亚细胞分布及荧光强度。结果单独二甲双胍不会导致LPS激活细胞发生焦亡,但二甲双胍呈剂量依赖性促进LPS+ATP诱导的细胞焦亡;免疫印迹法显示,在蛋白水平上,单独LPS(或LPS+二甲双胍)刺激不能诱导细胞释放成熟IL~(-1)β(17 ku)到细胞外;当加入ATP刺激后,成熟IL~(-1)β和活化caspase-1(10 ku)释放到胞外;而二甲双胍可以剂量依赖性地促进LPS激活、ATP刺激下腹腔巨噬细胞释放成熟IL~(-1)β和活化caspase-1的水平。结论二甲双胍增强LPS激活的腹腔巨噬细胞在ATP刺激下的炎症小体活化,提高caspase-1活化和成熟IL~(-1)β释放,促进炎症反应。
[Abstract]:Objective to investigate the effect of metformin, a common diabetic drug, on the activation of inflammatory corpuscles induced by extracellular ATP and the release of interleukin 1 尾 (IL 1) 尾 in peritoneal macrophages activated by lipopolysaccharide (LPS). Methods C57BL/6 mice were injected intraperitoneally with thioglycollate (30 g / L) to induce a large number of peritoneal macrophages, the inflammatory bodies were activated by LPS ATP treatment, and the effect of metformin on the apoptosis of peritoneal macrophages induced by LPS ATP was detected by iodized propidium iodide (Pi) staining. Immunoblotting assay was used to detect the expression of ILF-1) 尾 and caspase-1 in the cell and supernatant, and immunofluorescence technique was used to detect the subcellular distribution and fluorescence intensity of P_2X_7 purinergic ATP receptor (7P2XS7R) in macrophages. Results Metformin alone did not cause pyrolysis of LPS activated cells, but metformin promoted the apoptosis induced by LPS ATP in a dose-dependent manner. LPS- (or LPS metformin) stimulation could not induce the cells to release mature ILP-1) 尾 -ku17) into the cells, but after the addition of ATP, the mature ILP-1) 尾 and activated caspase-1(10 Kul) were released to the extracellular cells. Metformin dose-dependently promoted the release of mature ILF-1 尾 and activated caspase-1 by ATP-activated peritoneal macrophages. Conclusion Metformin enhances the activation of inflammatory corpuscles stimulated by ATP, enhances the activation of caspase-1 and the release of mature ILF-1) 尾 in LPS activated peritoneal macrophages, and promotes the inflammatory response.
【作者单位】：暨南大学生命科学技术学院免疫生物学系;暨南大学生命科学技术学院细胞生物学系;
【基金】：国家自然科学基金资助项目(No 81173604,81373423)
【分类号】：R96

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