淫羊藿苷在肺泡上皮A549细胞的摄取与抗炎作用研究  

Uptake and Anti-inflammatory Effect of Icariin in Alveolar Epithelial A549 Cells

作　　者：赵迪[1,2,3] 董浩然 刘学芳[1,2,3] 李先贺 赵杰[1,2,3] 冯素香 Zhao Di;Dong Haoran;Liu Xuefang;Li Xianhe;Zhao Jie;Feng Suxiang(Henan University of Traditional Chinese Medicine,Zhengzhou 450046;Co-construction Collaborative Innovation Center for Chinese Medicine and Respiratory Diseases by Henan&Education Ministry of P.R.China,Zhengzhou 450046;Henan Key Laboratory of Chinese Medicine for Respiratory Disease,Zhengzhou 450046)

机构地区：[1]河南中医药大学中医药科学院,郑州450046 [2]呼吸疾病中医药防治省部共建协同创新中心,郑州450046 [3]河南中医药防治呼吸病重点实验室,郑州450046

出　　处：《中药药理与临床》2021年第4期18-22,共5页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金资助项目(编号:81904033);河南省中医药大学博士基金科研项目(编号:BSJJ-2016-01)。

摘　　要：目的:探索淫羊藿苷在肺泡上皮A549细胞的摄取与抗炎作用。方法:采集、培养肺泡上皮A549细胞,采用肿瘤坏死因子-α(TNF-α)刺激建立体外炎症模型,噻唑蓝法(MTT)检测细胞存活率,建立HPLC-MS法并测定细胞内淫羊藿苷的含量。酶联免疫法(ELISA)检测细胞上清中白细胞介素-8(IL-8)、基质金属蛋白酶9(MMP9)与金属蛋白酶组织抑制因子1(TIMP-1)含量。结果:TNF-α建立炎症模型可降低细胞存活率(P<0.05);淫羊藿苷在浓度为12.5μg/mL~100μg/mL条件下对A549细胞无显著毒性;摄取实验中,空白对照组与模型对照组细胞对淫羊藿苷摄取达峰时间(T_(max))分别为360 min、300 min(P<0.05);淫羊藿给药浓度与最大摄取量(I_(max))呈正相关(r空白=0.976,r模型=0.982),模型对照组I_(max)高于空白对照组(P<0.01);外排实验中,淫羊藿苷在模型对照组细胞中消除迅速,外排较快。与空白对照组细胞相比,模型给药组细胞中IL-8、MMP9与TIMP-1水平显著升高(P<0.01)、TIMP-1/MMP9比值明显降低(P<0.05);与模型给药组细胞相比,用药组细胞IL-8、MMP水平随药物浓度升高明显下降(P<0.05),TIMP-1/MMP9比值有升高趋势。结论:炎症模型A549细胞对淫羊藿苷摄取量较大,外排迅速;淫羊藿苷可提高A549细胞炎症模型存活率,改善IL-8、MMP水平和TIMP-1/MMP9比值。Objective:To explore the uptake and its anti-inflammatory effect of icariin in alveolar epithelial A549 cells. Methods: A549 cells were collected and cultured, and the in vitro inflammation model was established by tumor necrosis factor-α(TNF-α) stimulation. Cell viability was determined by MTT assay. The HPLC-MS method was established to measure the concentration of icariin in cells. The levels of interleukin-8(IL-8), matrix metalloproteinase 9(MMP9), and metalloproteinase tissue inhibitor 1(TIMP-1) were detected by ELISA. Results: Compared with the normal control group, the model group showed reduced cell viability(P<0.05). Icariin at 12.5 μg/mL~100 μg/mL possessed no significant toxicity on A549 cells. The results of the uptake test showed that the T_(max) of icariin in the normal control group and the model group was 360 min and 300 min(P<0.05), respectively. The icariin concentration was positively correlated with the I_(max)(r_(normal)=0.976 and rmodel=0.982). I_(max) of the model group was higher than that in the normal control group(P<0.01). The efflux test revealed that icariin was eliminated fast in the model group. Compared with the normal control group, the model group showed increased levels of IL-8, MMP9, and TIMP-1(P<0.01) and declining TIMP-1/MMP9(P<0.05). Compared with the model group, the groups with drug intervention showed declining levels of IL-8 and MMP with the increase in icariin concentration(P<0.05) and an increasing trend of TIMP-1/MMP9.Conclusion: The inflammation model A549 cells exhibit a large uptake and a fast efflux of icariin. Icariin can effectively increase the cell viability of inflammatory cells and increase the levels of IL-8, MMP, and TIMP-1/MMP9.

关 键 词：淫羊藿苷 Ⅱ型肺泡上皮细胞A549 炎症模型 摄取 

分 类 号：R285[医药卫生—中药学]