摘要
本研究探究柚皮素对肺炎链球菌(SP)诱导的肺泡上皮细胞凋亡及MAPK/NF-κB信号通路的影响。培养肺泡上皮细胞HPAEpiC,使用CCK-8检测柚皮素对HPAEpiC细胞的毒性。再次培养HPAEpiC细胞,分为对照组、模型组、阳性对照组、柚皮素组和抑制剂组,ELISA法检测炎症因子的表达水平;TUNEL检测细胞凋亡水平;Western blot检测凋亡及MAPK/NF-κB信号通路相关蛋白表达水平。结果显示,柚皮素浓度≤60μmol/L时,对HPAEpiC细胞无毒性(P>0.05)。SP诱导可上调细胞上清液中炎症因子(TNF-α和IL-1β)的释放量、细胞凋亡率和细胞中凋亡相关蛋白(Cleaved caspase-8、Cleaved caspase-9和Cleaved caspase-3)与MAPK/NF-κB信号通路相关蛋白(p-JNK、p-p38、p-ERK、p-p65和p-IκBα)的表达水平。柚皮素干预后可改善SP对肺泡上皮细胞的影响,且添加MAPK/NF-κB信号通路抑制剂干预与柚皮素干预对SP诱导的肺泡上皮细胞的作用效果具有相似性。以上结果提示柚皮素可能通过抑制MAPK/NF-κB信号通路减弱SP诱导的肺泡上皮细胞凋亡和炎症反应。
This study was to explore the effects of naringenin on the alveolar epithelial cells apoptosis and MAPK/NF-κB signaling pathway induced by Streptococcus pneumoniae(SP).The alveolar epithelial cells HPAEpiC were cultured, and CCK-8 was used to detect the toxicity of naringenin to HPAEpiC cells.HPAEpiC cells were cultured again, and they were divided into control group, model group, positive control group, naringenin group and inhibitor group.ELISA method was used to detect the expression level of inflammatory factors.TUNEL was used to detect the level of apoptosis.Western blot was used to detect the expression levels of apoptosis and MAPK/NF-κB signaling pathway related proteins.The results showed that naringenin was not toxic to HPAEpiC cells when the concentration of naringenin was ≤60 μmol/L(P>0.05).SP induction could up-regulate the release of inflammatory factors(TNF-α and IL-1β) in the cell supernatant, the rate of apoptosis, and the expression levels of apoptosis-related proteins(Cleaved caspase-8,Cleaved caspase-9 and Cleaved caspase-3) in the cell, and MAPK/NF-κB signaling pathway related proteins(p-JNK,p-p38,p-ERK,p-p65 and p-IκBα).Naringenin intervention could improve the effects of SP on alveolar epithelial cells, and the effect of adding MAPK/NF-κB signaling pathway inhibitors and naringenin intervention on SP-induced alveolar epithelial cells was similar.The above results suggest that naringenin may attenuate the apoptosis and inflammatory response of alveolar epithelial cells induced by SP through inhibiting the MAPK/NF-κB signaling pathway.
作者
陈运庭
王睿雯
麦明志
杨国通
CHEN Yun-ting;WANG Rui-wen;MAI Ming-zhi;YANG Guo-tong(Respiratory Medicine,Sanya People's Hospital,Sanya 572000,China)
出处
《药物生物技术》
CAS
2023年第1期19-24,共6页
Pharmaceutical Biotechnology
基金
海南省卫生计生行业科研项目(No.1802330243A2001)。
