淫羊藿苷对香烟烟雾提取物诱导上皮细胞炎症的保护作用  

Protective effect of icariin on cigarette smoke extract induced inflammation in alveolar epithelial cells

作　　者：曾丽璇[1] 弓唯一[1] 张红英[1] 孙婧[1] 罗清莉[1] 段晓红[1] 董竞成[1] 

机构地区：[1]复旦大学中西医结合研究所,上海200040

出　　处：《中华中医药杂志》2013年第7期2114-2118,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家重点基础研究发展计划(973计划)资助项目(No.2009CB523001);2009年度上海市优秀学科带头人计划(A类)项目(No.09XD1400700);上海市卫生局中医药科研基金(No.2008L018A)~~

摘　　要：目的:探讨淫羊藿苷对香烟烟雾提取物(CSE)诱导的上皮细胞炎症的影响。方法:CSE刺激SD乳鼠肺泡上皮细胞(ATⅡ)建立细胞模型。细胞分6组:对照组、CSE组、PDTC[核因子KappaB(NF-κB)抑制剂]组、淫羊藿苷低(40μg/mL)、中(80μg/mL)和高(160μg/mL)剂量组。改良巴氏染色和电镜鉴定ATⅡ细胞;CCK-8法检测药物对细胞增殖的影响;ELISA法检测细胞上清液中细胞因子含量;蛋白质印迹法检测I Kappa B-α(IκB-α)和P-p65磷酸化水平;凝胶迁移实验(EMSA)检测NF-κB的DNA结合活力。结果:分离培养的细胞巴氏染色阳性;淫羊藿苷各组均未显示出细胞毒性,CSE对ATⅡ细胞呈现出时间和浓度依赖性抑制;CSE组细胞上清液中白细胞介素-8(IL-8)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)较对照组显著性升高(P<0.05),IL-10显著性降低(P<0.05),淫羊藿苷组随着剂量增高IL-8、IL-6、TNF-α表达逐渐降低,IL-10表达增加。经CSE诱导后,P-p65蛋白表达较对照组增加(P<0.05),IκB-α则减少(P<0.05);淫羊藿苷干预后,P-p65表达减少(P<0.05),IκB-α增多(P<0.05)。对照组NF-κB的DNA结合活力较弱,CSE组显著性增强(P<0.05),淫羊藿苷干预后减弱(P<0.05)。结论:淫羊藿苷抑制CSE诱导的ATⅡ细胞IL-8、IL-6、TNF-α表达和NF-κB活化,促进IL-10表达。Objective： To investigate the effect of icariin on cigarette smoke extract（CSE） induced inflammation in alveolar epithelial（AT II） cells.Methods： Cell model was established by stimulating AT II of neonatal SD rats by CSE.The cells were divided into six groups： control group,CSE group,PDTC group,40μg/mL,80μg/mL and 160μg/mL icariin group.AT II cells were identified by modified papanicolaou staining and electron microscopy;cell proliferation was detected by CCK-8 assay;cytokines in cell supernatants was measured by ELISA assay;expressions of IκB-α and p65 phosphorylation were identified by western blot;NF-κB DNA binding activity was detected by electrophoretic mobility shift assay（EMSA）.Results： Isolated and cultured cells were positive by papanicolaou stain;icariin groups did not show cytotoxicity,while CSE showed a time and concentration-dependent inhibition on AT II cells;compared with control group,IL-8,IL-6 and TNF-α in cell supernatant in CSE group significantly increased,with IL-10 decreased,while as the dose increased,IL-8,IL-6,TNF-α expression icariin groups decreased,with IL-10 increased.After CSE stimulation,P-p65 expression increased and IκB-α decreased;P-p65 expression decreased and IκB-α increased in icariin groups.NF-κB DNA binding activity was weak in control group,and was enhanced significantly in CSE group.After icariin intervention,it was weakened.Conclusion： The Icariin attenuated CSE-induced expression of IL-8,IL-6,TNF-α and NF-κB activation,and increased IL-10 expression in AT II cells.

关 键 词：淫羊藿苷 香烟烟雾提取物 上皮细胞炎症 核因子-ΚB 

分 类 号：R285.5[医药卫生—中药学]