羟基红花黄色素A对香烟提取物诱导A549细胞炎症因子表达作用的研究  

Hydroxysafflor yellow a inhibits inflammatory factors expression elevation induced by cigarette smoke extract in A549 human alveolar epithelial cells

作　　者：薛长江[1,2] 董芳[1,2] 王宇[1,2] 臧宝霞[1,2] 金鸣[1,2] 

机构地区：[1]首都医科大学附属北京安贞医院 [2]北京市心肺血管研究所药理研究室,北京100029

出　　处：《心肺血管病杂志》2014年第2期276-280,共5页Journal of Cardiovascular and Pulmonary Diseases

基　　金：国家自然科学基金科学基金资助项目(81270103)

摘　　要：目的:探讨羟基红花黄色素A(HSYA)抑制香烟烟雾提取物(CSE)诱导的A549肺泡上皮细胞炎症介质表达升高的作用。方法:用CSE刺激A549肺泡上皮细胞建立损伤模型,细胞分七组:单纯HSYA组、Sham组、CSE组、CSE+HSYA低剂量组(1μmol/L)、CSE+HSYA中剂量组(4μmol/L)、CSE+HSYA高剂量组(16μmol/L)和CSE+阳性对照药红霉素(5μg/mL)组。以RT-qPCR技术检测白介素(IL)-6、白介素(IL)-1β、肿瘤坏死因子(TNF)-α、细胞间黏附分子(ICAM)-1和血管细胞黏附分子(VCAM)-1 mRNA的表达水平;ELISA法检测细胞培养液中IL-6、IL-1β及TNF-α的蛋白水平表达;蛋白质印迹法(western blot)检测细胞p38MAPK磷酸化的水平。结果:与Sham组相比,CSE组IL-6、IL-1β、TNF-α、ICAM-1和VCAM-1 mRNA水平均明显升高(P<0.001),给予不同浓度的HSYA后炎症因子的高表达受到抑制,红霉素组也见较明显的抑制。与Sham组相比,CSE组细胞上清液中IL-6、IL-1β和TNF-α蛋白的表达水平明显升高(P<0.001),给予不同浓度的HSYA后CSE所诱发的炎症因子的高表达受到抑制,红霉素组也见较明显的抑制。与Sham组比较,CSE组的p38 MAPK的磷酸化水平明显升高(P<0.001),给予不同浓度的HSYA后细胞p38 MAPK的磷酸化水平升高受到抑制,红霉素组也可见明显的抑制。结论:HSYA可抑制CSE诱导A549细胞的炎症因子的表达升高。Objective：To study the inhibitory effect of HSYA on cigarette smoke extract （CSE） induced A549 human alveolar epithelial cell inflammatory factor expression elevation.Methods：CSE induced A549 cell inflammatory injury model was established and cells were divided into seven groups including HSYA blank group,Sham group,CSE group,CSE ＋ low HSYA group （1 μmol/L）,CSE ＋ middle HSYA group （4 μmol/L）,C SE ＋ high HSYA group （16 μmol/L） and CSE ＋ Erythromycin group （ERY,5 μg/mL）as positive control.Various mRNA levels in A549 cell were measured by RT-qPCR technology.Inflammatory factors protein levels in cell culture media were detected by ELISA assay.Western blot analysis was used to determine phosphorylated p38 MAPK level in A549 cells.Results：HSYA （1 μmol/L,4μmol/L or 16μmol/L） attenuated IL-6,IL-1β,TNF-α,VCAM-1 and ICAM-1 mRNA expression elevation induced by CSE （CSE group vs.Sham group,P ＜ 0.001 ; CSE ＋ HSYA 4 μmol/L or CSE ＋ 16μmol/L group vs.CSE group,P ＜ 0.05 or P ＜ 0.01）.The endothelium IL-6,IL-1β or TNF-α protein expression increased after CSE （15％） addition and HSYA can attenuate these changes.（CSE group vs.Sham group,P ＜0.001 ; CSE ＋ HSYA group vs.CSE group,P ＜0.05 or P ＜ 0.01）.HSYA treatment also inhibited phosphorylation of p38 mitogen-activated protein kinase （p38MAPK） induced by CSE in A549 cell.Conclusion：It is suggested that HSYA is active to inhibit CSE induced A549 cell inflammatory factor expression elevation and p38MAPK phosphorylation.

关 键 词：羟基红黄色素A 香烟烟雾提取物 炎症因子 p38 MAPK 

分 类 号：R54[医药卫生—心血管疾病]