【摘要】：目的:探討脂多糖(lipopolysaccharide,LPS)聯(lián)合真菌葡聚糖能否活化支氣管上皮細(xì)胞16HBE內(nèi)NOD樣受體熱蛋白結(jié)構(gòu)域相關(guān)蛋白3(Nod-like receptor pyrin domain-containing protein 3,NLRP3)炎性小體,芍藥苷(paeoniflorin,PF)對(duì)該NLRP3炎性小體的活化是否有抑制作用及相關(guān)機(jī)制。方法:LPS聯(lián)合真菌葡聚糖建立感染模型,RT-PCR檢測(cè)細(xì)胞內(nèi)NLRP3、caspase-1、白細(xì)胞介素(interleukin,IL)-1βm RNA的表達(dá);ELISA檢測(cè)細(xì)胞上清中IL-1β含量;caspase-1活性檢測(cè)試劑盒檢測(cè)胞內(nèi)caspase-1活化程度;流式檢測(cè)胞內(nèi)活性氧(reactive oxygen species,ROS)變化;Western blot檢測(cè)胞內(nèi)NLRP3、caspase-1、IL-1β蛋白表達(dá)變化。結(jié)果:LPS聯(lián)合真菌葡聚糖聯(lián)合作用于支氣管上皮細(xì)胞,胞內(nèi)NLRP3、caspase-1、IL-1β表達(dá)轉(zhuǎn)錄加強(qiáng),細(xì)胞上清中IL-1β含量增加,caspase-1活性上調(diào),細(xì)胞內(nèi)ROS升高;PF預(yù)作用細(xì)胞后,胞內(nèi)ROS隨著藥物濃度增加而逐漸降低,NLRP3、caspase-1、IL-1β的轉(zhuǎn)錄表達(dá)也隨之受抑制而下調(diào)。結(jié)論:LPS聯(lián)合真菌葡聚糖可有效活化支氣管上皮細(xì)胞內(nèi)NLRP3炎性小體,而PF能有效抑制胞內(nèi)ROS產(chǎn)生從而抑制炎性小體活化、抑制真菌葡聚糖所致的炎性反應(yīng)。
[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.
【作者單位】： 南京醫(yī)科大學(xué)第一附屬醫(yī)院呼吸科;
【基金】：江蘇省呼吸病臨床醫(yī)學(xué)研究中心項(xiàng)目(BL2012012)
【分類號(hào)】：R519

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本文編號(hào)：2188167