芍药苷减弱真菌葡聚糖诱导的支气管上皮细胞内NLRP3炎性小体活化
发布时间：2018-08-17 16:02
【摘要】：目的:探讨脂多糖(lipopolysaccharide,LPS)联合真菌葡聚糖能否活化支气管上皮细胞16HBE内NOD样受体热蛋白结构域相关蛋白3(Nod-like receptor pyrin domain-containing protein 3,NLRP3)炎性小体,芍药苷(paeoniflorin,PF)对该NLRP3炎性小体的活化是否有抑制作用及相关机制。方法:LPS联合真菌葡聚糖建立感染模型,RT-PCR检测细胞内NLRP3、caspase-1、白细胞介素(interleukin,IL)-1βm RNA的表达;ELISA检测细胞上清中IL-1β含量;caspase-1活性检测试剂盒检测胞内caspase-1活化程度;流式检测胞内活性氧(reactive oxygen species,ROS)变化;Western blot检测胞内NLRP3、caspase-1、IL-1β蛋白表达变化。结果:LPS联合真菌葡聚糖联合作用于支气管上皮细胞,胞内NLRP3、caspase-1、IL-1β表达转录加强,细胞上清中IL-1β含量增加,caspase-1活性上调,细胞内ROS升高;PF预作用细胞后,胞内ROS随着药物浓度增加而逐渐降低,NLRP3、caspase-1、IL-1β的转录表达也随之受抑制而下调。结论:LPS联合真菌葡聚糖可有效活化支气管上皮细胞内NLRP3炎性小体,而PF能有效抑制胞内ROS产生从而抑制炎性小体活化、抑制真菌葡聚糖所致的炎性反应。
[Abstract]:Objective: to investigate whether lipopolysaccharide (LPS) combined with fungal dextran can activate the inflammatory body of NOD like receptor heat domain associated protein 3 (Nod-like receptor pyrin domain-containing protein 3) in 16HBE of bronchial epithelial cells. Whether paeoniflorin PF can inhibit the activation of NLRP3 inflammatory corpuscles and its related mechanisms. Methods RT-PCR was used to detect the expression of NLRP3caspase-1 and interleukin (IL) -1 尾 m RNA in the supernatant of the supernatant. The activity of IL-1 尾 in the supernatant was detected by Elisa. The activity of caspase-1 in the supernatant was detected by Elisa. The activity of caspase-1 in the supernatant was detected by RT-PCR. The changes of intracellular reactive oxygen species (reactive oxygen speciesus Ros) were detected by flow cytometry and the expression of IL-1 尾 protein in NLRP3Caspase-1 was detected by Western blot. Results the expression of IL-1 尾 of NLRP3caspase-1 was enhanced in bronchial epithelial cells induced by the combination of IL-1 尾 and fungal dextran, and the activity of caspase-1 increased in the supernatant of the cells. The intracellular ROS increased the expression of IL-1 尾 in the pretreated cells of PF. The transcriptional expression of NLRP3 caspase-1 and IL-1 尾 was inhibited and down-regulated with the increase of drug concentration. Conclusion NLRP3 inflammatory corpuscles in bronchial epithelial cells can be effectively activated by the combination of NLRP3 and fungal dextran, while PF can effectively inhibit the production of intracellular ROS, inhibit the activation of inflammatory bodies and inhibit the inflammatory response induced by fungal dextran.
【作者单位】： 南京医科大学第一附属医院呼吸科;
【基金】：江苏省呼吸病临床医学研究中心项目(BL2012012)
【分类号】：R519
[Abstract]:Objective: to investigate whether lipopolysaccharide (LPS) combined with fungal dextran can activate the inflammatory body of NOD like receptor heat domain associated protein 3 (Nod-like receptor pyrin domain-containing protein 3) in 16HBE of bronchial epithelial cells. Whether paeoniflorin PF can inhibit the activation of NLRP3 inflammatory corpuscles and its related mechanisms. Methods RT-PCR was used to detect the expression of NLRP3caspase-1 and interleukin (IL) -1 尾 m RNA in the supernatant of the supernatant. The activity of IL-1 尾 in the supernatant was detected by Elisa. The activity of caspase-1 in the supernatant was detected by Elisa. The activity of caspase-1 in the supernatant was detected by RT-PCR. The changes of intracellular reactive oxygen species (reactive oxygen speciesus Ros) were detected by flow cytometry and the expression of IL-1 尾 protein in NLRP3Caspase-1 was detected by Western blot. Results the expression of IL-1 尾 of NLRP3caspase-1 was enhanced in bronchial epithelial cells induced by the combination of IL-1 尾 and fungal dextran, and the activity of caspase-1 increased in the supernatant of the cells. The intracellular ROS increased the expression of IL-1 尾 in the pretreated cells of PF. The transcriptional expression of NLRP3 caspase-1 and IL-1 尾 was inhibited and down-regulated with the increase of drug concentration. Conclusion NLRP3 inflammatory corpuscles in bronchial epithelial cells can be effectively activated by the combination of NLRP3 and fungal dextran, while PF can effectively inhibit the production of intracellular ROS, inhibit the activation of inflammatory bodies and inhibit the inflammatory response induced by fungal dextran.
【作者单位】： 南京医科大学第一附属医院呼吸科;
【基金】：江苏省呼吸病临床医学研究中心项目(BL2012012)
【分类号】：R519
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1 黄燕华;华檬;崔学范;;芍药苷减弱真菌葡聚糖诱导的支气管上皮细胞内NLRP3炎性小体活化[J];南京医科大学学报(自然科学版);2017年03期
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