基于NF-κB信号通路的玉屏风散加味含药血清对RLE-6TN细胞炎症反应的影响  

Effects of Modified Yupingfeng Powder Medicated Serum on RLE-6TN Cells Inflammatory Reaction Based on NF-κB Signaling Pathway

作　　者：孟佳磊 马宇慧 袁林[1] 叶慧 华旭 雷鸣[1] MENG Jialei;MA Yuhui;YUAN Lin;YE Hui;HUA Xu;LEI Ming(Seventh People’s Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200137,China;Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学附属第七人民医院,上海200137 [2]上海中医药大学,上海201203

出　　处：《中国中医药信息杂志》2024年第1期84-90,共7页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(82174189);急救与创伤研究教育部重点实验室(海南医学院)开放课题基金项目(KLET-202115);浦东新区科技发展基金民生科研专项(PKJ2021-Y06);浦东新区卫生健康委员会学科建设计划(PWZxq2022-09)。

摘　　要：目的观察玉屏风散加味含药血清对大鼠肺上皮Ⅱ型细胞RLE-6TN炎症反应的影响,基于核因子(NF)-κB信号通路探讨其作用机制。方法采用脂多糖诱导RLE-6TN细胞炎症反应,将细胞分为对照组、模型组、地塞米松血清组和5%、10%、20%中药血清组,分别以不同血清干预细胞24 h。试剂盒检测细胞上清液乳酸脱氢酶(LDH)含量,荧光染色检测细胞活性氧(ROS)含量,Hoechst/PI双染观察细胞凋亡情况,ELISA检测细胞上清液肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6、IL-1β含量,RT-qPCR检测细胞Toll样受体4(TLR4)、髓样分化因子88(MyD88)、NF-κB抑制蛋白α(IκBα)mRNA表达,Western blot检测细胞TLR4、MyD88、NF-κBp65、IκBα蛋白表达。结果与对照组比较,模型组RLE-6TN细胞上清液LDH、TNF-α、IL-6、IL-1β及细胞ROS含量显著升高(P<0.01),细胞凋亡率显著升高(P<0.01),TLR4、MyD88 mRNA和蛋白表达显著升高(P<0.01),IκBαmRNA表达显著降低(P<0.01),p-NF-κBp65/NF-κBp65、p-IκBα/IκBα比值显著升高(P<0.01);与模型组比较,20%中药血清组和地塞米松血清组细胞RLE-6TN细胞上清液LDH、TNF-α、IL-6、IL-1β及细胞ROS含量显著降低(P<0.01),细胞凋亡率显著降低(P<0.01),TLR4、MyD88 mRNA和蛋白表达显著降低(P<0.05,P<0.01),IκBαmRNA表达显著升高(P<0.01),p-NF-κBp65/NF-κBp65、p-IκBα/IκBα比值显著降低(P<0.05,P<0.01)。结论玉屏风散加味含药血清可减轻RLE-6TN细胞炎症反应,其机制可能与调控NF-κB信号通路有关。Objective To investigate the effects of modified Yupingfeng Powder medicated serum on the inflammatory response of rat lung epithelial typeⅡcells RLE-6TN;To explore the mechanism based on nuclear factor(NF)-κB signaling pathway.Methods The inflammatory reaction of RLE-6TN cells was induced by lipopolysaccharide,and the cells were divided into control group,model group,dexamethasone medicated serum group,and 5%,10%and 20%TCM serum group,and the cell model was intervened with different serum.After 24 hours,the content of lactate dehydrogenase(LDH)in cell supernatant was detected by kit,and the content of reactive oxygen species(ROS)in cells were detected by fluorescence staining,Hoechst/PI double staining was used to observe cell apoptosis,and ELISA was used to detect tumor necrosis factor-α(TNF-α),interleukin(IL)-6,and IL-1βcontents in cell supernatant,RT-qPCR was used to detect Toll like receptor 4(TLR4),myeloid differentiation factor 88(MyD88),and NF-κB inhibitory proteinα(IκBα)mRNA expression,Western blot was used to detect protein expressions of TLR4,MyD88,NF-κBp65 and IκBα.Results Compared with the control group,the contents of LDH,TNF-α,IL-6,IL-1βin RLE-6TN cell supernatant and ROS content in cells of model group significantly increased(P<0.01),the cell apoptosis rate significantly increased(P<0.01),the expressions of TLR4 and MyD88 mRNA and protein significantly increased(P<0.01),the expression of IκBαmRNA significantly decreased(P<0.01),and the ratio of p-NF-κBp65/NF-κBp65 and p-IκBα/IκBαsignificantly increased(P<0.01).Compared with the model group,the content of LDH,TNF-α,IL-6,IL-1 in RLE-6TN cell supernatant and ROS content in cells significant decreased in the 20%TCM serum group and dexamethasone medicated serum group(P<0.01),the cell apoptosis rate significantly decreased(P<0.01),the expressions of TLR4 and MyD88 mRNA and protein significantly decreased(P<0.05,P<0.01),the expression of IκBαmRNA significantly increased(P<0.01),and the ratio of p-NF-κBp65/NF-κBp65 and p-IκBα

关 键 词：玉屏风散加味 脓毒症急性肺损伤 含药血清 NF-ΚB信号通路 RLE-6TN细胞 

分 类 号：R285.5[医药卫生—中药学]