摘要
目的:探讨松果菊苷(echinacoside,ECH)对博来霉素(bleomycin,BLM)诱导的肺上皮细胞损伤、炎症反应及核因子E2相关因子2(Nrf-2)、核因子κB(NF-κB)p65和环氧合酶-2(COX-2)蛋白表达的影响。方法:分别以1、2.5、5和10 mg/L的BLM及5、25、125和625 mg/L的ECH处理体外培养的人Ⅱ型肺泡上皮细胞系HEPApiC以探索BLM和ECH的最佳实验浓度。根据筛选的BLM和ECH的最佳实验浓度,将HEPApiC细胞分为对照组、BLM组和BLM+ECH组,每组均进行3次独立实验。CCK-8法检测细胞活力;Hoechst 33258染色法和流式细胞术检测细胞凋亡情况;ELISA法检测炎症因子肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)和IL-1β的水平;试剂盒法测定丙二醛(MDA)、活性氧簇(ROS)和超氧化物歧化酶(SOD)的水平;Western blot法检测Nrf-2、NF-κB p65和COX-2的蛋白表达水平。结果:BLM和ECH的最佳实验浓度分别为10 mg/L和125 mg/L。与对照组比较,BLM组细胞活力、SOD活性及细胞质Nrf-2和细胞质NF-κB p65的水平均显著降低(P<0.05,P<0.01),而细胞凋亡水平、TNF-α、IL-6、IL-1β、ROS、MDA及COX-2、细胞核Nrf-2和细胞核NF-κB p65的水平均显著增加(P<0.01);与BLM组比较,BLM+ECH组细胞活力、SOD活性及细胞核Nrf-2和细胞质NF-κB p65水平均显著增加(P<0.01),而细胞凋亡水平、TNF-α、IL-6、IL-1β、ROS、MDA及COX-2、细胞质Nrf-2和细胞核NF-κB p65的水平均显著降低(P<0.05,P<0.01)。结论:在BLM处理的HEPApiC细胞中,ECH能抑制氧化应激损伤和炎症反应,且可减少NF-κB核转位和降低COX-2蛋白表达,并增加Nrf-2核转位。
AIM:To investigate the effects of echinacoside(ECH)on bleomycin(BLM)-induced lung epithe-lial cell injury,inflammatory response,and the protein expression of nuclear factor E2-related factor 2(Nrf-2),nuclear factor-κB(NF-κB)p65 and cyclooxygenase-2(COX-2).METHODS:Human typeⅡalveolar epithelial cells(HEPApiCs)were treated with 1,2.5,5 and 10 mg/L BLM,and 5,25,125 and 625 mg/L ECH,respectively,to explore the optimal ex-perimental concentrations of BLM and ECH.According to the optimal concentration of BLM and ECH,HEPApiCs were di-vided into control group,BLM group and BLM+ECH group,and 3 independent experiments were performed for each group.The viability was measured by CCK-8 assay.The apoptosis was detected by Hoechst 33258 staining and flow cytom-etry.The levels of inflammatory cytokines tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and IL-1βwere mea-sured by ELISA.The levels of malondialdehyde(MDA),reactive oxygen species(ROS)and superoxide dismutase(SOD)were also determined.The protein expression levels of Nrf-2,NF-κB p65 and COX-2 were detected by Western blot.RESULTS:The optimal concentrations of BLM and ECH were 10 mg/L and 125 mg/L,respectively.Compared with control group,the viability,SOD activity,and protein levels of cytoplasmic Nrf-2 and cytoplasmic NF-κB p65 in BLM group were significantly decreased(P<0.05,P<0.01),while,apoptosis rate,TNF-α,IL-6,IL-1β,ROS,MDA,COX-2,nuclear Nrf-2 and nuclear NF-κB p65 levels were significantly increased(P<0.01).Compared with BLM group,the viability,SOD activity,and protein levels of nuclear Nrf-2 and cytoplasmic NF-κB p65 levels in BLM+ECH group were significantly increased(P<0.01).The levels of apoptosis rate,TNF-α,IL-6,IL-1β,ROS,MDA,COX-2,cytoplasmic Nrf-2 and nuclear NF-κB p65 were significantly decreased(P<0.05,P<0.01).CONCLUSION:ECH inhibits oxidative stress injury and inflammatory response,reduces the nuclear translocation of NF-κB and decreases COX-2 protein expres-sion,and increases the nuclear translocation of Nrf-2 in BLM-treated HEPApiCs.
作者
崔艳红
李克芳
金博
彭飞
刘培培
曲伟伟
门翔
赵江
CUI Yan-hong;LI Ke-fang;JIN Bo;PENG Fei;LIU Pei-pei;QU Wei-wei;MEN Xiang;ZHAO Jiang(The Second Ward,Department of Respiratory and Critical Care Medicine,Nanyang Central Hospital,Nanyang 473000,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2021年第11期2024-2030,共7页
Chinese Journal of Pathophysiology
基金
河南省2018年度科技发展计划项目(No.182102310184)。
关键词
松果菊苷
博来霉素
肺上皮细胞
氧化应激
炎症反应
Echinacoside
Bleomycin
Lung epithelial cells
Oxidative stress
Inflammatory response
