橙皮苷抑制ROS/NLRP3通路对肺炎链球菌感染的肺泡上皮细胞损伤的影响OA
Effect of hesperidin on alveolar epithelial cell damage caused by Streptococcus pneumoniae infection by inhibiting ROS/NLRP3 pathway
目的:探究橙皮苷抑制活性氧(ROS)/核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)通路对肺炎链球菌感染的肺泡上皮细胞损伤的影响.方法:将肺泡上皮细胞A549分为对照组(不作处理)、感染组(1×108 CFU/mL肺炎链球菌刺激细胞)、感染+低、中、高剂量橙皮苷组(感染后分别使用30、60、90 µmol/L橙皮苷处理)、感染+高剂量橙皮苷+通路激活剂三甲胺N-氧化物(TMAO)组(感染后使用90 µmol/L橙皮苷和200 µmol/L TMAO共同处理).CCK-8法检测细胞活力;流式细胞术分析细胞凋亡情况;ELISA检测细胞炎症因子TNF-α、IL-1β和IL-6水平;DCFH-DA荧光探针检测细胞ROS水平;试剂盒检测细胞MDA含量及CAT、SOD活性;Western blot分析细胞Ki67、半胱天冬酶3(caspase-3)、诱生型环氧化酶(Cox-2)、烟酰胺腺嘌呤二核苷酸磷酸氧化酶(Nox)和NLRP3蛋白水平.结果:与对照组比较,感染组细胞A450值、CAT和SOD活性、Ki67蛋白水平降低(P<0.05),细胞凋亡率、TNF-α、IL-1β、IL-6和MDA水平、ROS荧光强度、caspase-3、Nox和NLRP3蛋白水平升高(P<0.05);与感染组比较,感染+低、中、高剂量橙皮苷组细胞A450值、CAT和SOD活性、Ki67蛋白水平升高(P<0.05),细胞凋亡率、TNF-α、IL-1β、IL-6和MDA水平、ROS荧光强度、caspase-3、Nox和NLRP3蛋白水平降低(P<0.05);与感染+高剂量橙皮苷组比较,感染+高剂量橙皮苷+TMAO组细胞A450值、CAT和SOD活性、Ki67蛋白水平降低(P<0.05),细胞凋亡率、TNF-α、IL-1β、IL-6和MDA水平、ROS荧光强度、caspase-3、Nox和NLRP3蛋白水平升高(P<0.05).结论:橙皮苷可能通过抑制ROS/NLRP3通路减轻肺炎链球菌感染的肺泡上皮细胞损伤.
Objective:To investigate effect of hesperidin on alveolar epithelial cell damage caused by Streptococcus pneumoniae infection by inhibiting reactive oxygen species(ROS)/nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)pathway.Methods:A549 alveolar epithelial cells were assigned into control group(untreated),infection group(1×108 CFU/mL Strep-tococcus pneumoniae stimulated cells),infection+low,medium and high-doses hesperidin groups(treated with 30,60,and 90 µmol/L hesperidin after infection),and infection+high-dose hesperidin+pathway activator trimethylamine N-oxide(TMAO)group(treated with 90 µmol/L hesperidin and 200 µmol/L TMAO after infection).CCK-8 assay was used to detect cell viability.Flow cytometry was applied to analyze cell apoptosis.ELISA was applied to detect levels of cytokines TNF-α,IL-1β and IL-6 in cells.DCFH-DA fluores-cent probe was applied to detect cellular ROS level.Reagent kit was used to detect MDA content and CAT,SOD activities in cells.Western blot was applied to analyze levels of Ki67,caspase-3,cyclooxygenase-2(Cox-2),nicotinamide adenine dinucleotide phos-phate oxidase(Nox)and NLRP3 proteins in cells.Results:Compared with control group,A450 value,CAT and SOD activities,and Ki67 protein level in infection group were decreased(P<0.05),while apoptosis rate,TNF-α,IL-1β,IL-6 and MDA levels,ROS fluo-rescence intensity,caspase-3,Nox and NLRP3 protein levels were increased(P<0.05).Compared with infection group,A450 value,CAT and SOD activities,and Ki67 protein level in infection+low/medium/high-dose hesperidin groups were increased(P<0.05),while apoptosis rate,TNF-α,IL-1β,IL-6 and MDA levels,ROS fluorescence intensity,caspase-3,Nox and NLRP3 protein levels were decreased(P<0.05).Compared with infection+high-dose hesperidin group,A450 value,CAT and SOD activities and Ki67 protein level ininfection+high-dose hesperidin+TMAO group were decreased(P<0.05),while apoptosis rate,TNF-α,IL-1β,IL-6 and MDA levels,ROS fluorescence intensity,caspase-3,Nox and NLRP3 protein levels were increased(P<0.05).Conclusion:Hesperidin may alleviate alveolar epithelial cell damage caused by Streptococcus pneumoniae infection by inhibiting ROS/NLRP3 pathway.
黄文静;杨向琪;张艳敏;任乾;曾晓兵
张掖市第二人民医院儿科,张掖 734000张掖市第二人民医院儿科,张掖 734000张掖市第二人民医院儿科,张掖 734000张掖市第二人民医院儿科,张掖 734000张掖市第二人民医院儿科,张掖 734000
医药卫生
橙皮苷活性氧核苷酸结合寡聚化结构域样受体蛋白3肺炎链球菌肺泡上皮细胞损伤
HesperidinReactive oxygen speciesNucleotide-binding oligomerization domain-like receptor protein 3Strepto-coccus pneumoniaeAlveolar epithelial cell damage
《中国免疫学杂志》 2026 (3)
541-546,6
甘肃省科技计划项目(21JR1RG311)2023年甘肃省卫生健康行业科研项目(GSWSQN2023-19).
评论