淫羊藿苷对补体旁路激活诱导的小鼠急性肺损伤的保护作用  

Effects of icariin on acute lung injury in mice induced by activation of the complement alternative pathway

作　　者：郭静[1,2] 李娇[1,2] 张霖 杨毅[2] 朱芳娟[1,2] 李敏[3] 孙黔云 GUO Jing;LI Jiao;ZHANG Lin;YANG Yi;ZHU Fang-juan;LI Min;SUN Qian-yun(State Key Laboratory of Functions and Applications of Medicinal Plants,Guizhou Medical University,Guiyang 550014,China;Center for Pharmacology and Drug Screening,Key Laboratory of Chemistry for Natural Products,Guizhou Province and Chinese Acade)

机构地区：[1]贵州医科大学省部共建药用植物功效与利用国家重点实验室,贵州贵阳550014 [2]贵州省中国科学院天然产物化学重点实验室药理与生物活性研究中心,贵州贵阳550014 [3]贵州省人民医院干医科,贵州贵阳550002

出　　处：《中国药理学通报》2018年第7期988-993,共6页Chinese Pharmacological Bulletin

基　　金：贵州省科技计划项目(No 黔科合基础[2017]1115);贵州省科技创新人才团队项目(No 黔科合平台人才[2016]5625);贵州省高层次创新型人才培养项目(百层次)(No 黔科合人才[2016]4018)

摘　　要：目的探讨淫羊藿苷(ICA)对补体旁路激活造成的小鼠急性肺损伤的保护作用及作用机制。方法 32只昆明小鼠随机分为正常对照组、模型组、PDTC组、淫羊藿苷组,预防给药7 d后,用特异性补体旁路激活蛋白眼镜蛇毒因子(CVF),尾静脉注射复制小鼠急性肺损伤模型。测定小鼠支气管肺泡灌洗液(BALF)中蛋白含量和炎性细胞数目;肺组织匀浆后测定髓过氧化物酶(MPO)活性;ELISA法检测BALF和血清中IL-6、TNF-α、P-selectin、ICAM-1的含量;HE染色法观察肺组织病理形态学变化;免疫组化法测定肺组织中NF-κB p65的磷酸化水平,并采用双萤光素酶报告基因检测系统,在细胞水平上测定补体旁路激活对微血管内皮细胞中NF-κB的核内转录活性的影响。结果 ICA可以明显降低小鼠肺组织匀浆液中MPO含量和BALF中炎性细胞数目,并同时降低BALF中IL-6、TNF-α、P-selectin含量和血清中TNF-α、P-selectin、ICAM-1水平,减轻小鼠肺部炎性细胞浸润,明显抑制小鼠肺组织中NF-κB p65的磷酸化,并有效抑制NF-κB核内转录活性的上调。结论 ICA能有效减轻补体旁路激活诱导的小鼠肺部急性炎症反应,其机制可能与抑制肺组织炎性细胞浸润、NF-κB p65的磷酸化及核内转录活性,从而降低炎症反应水平有关。Aim To explore the protective effect and mechanism of icariin（ ICA） on acute lung injury（ ALI） in mice induced by activation of the complement alternative pathway. Methods 32 healthy KM mice were randomly divided into four groups： the normal group,the model group,the PDTC group and the Icariin group,which received 7-day intragastric administration respectively. Then cobra venom factor（ CVF）was used to activate specifically complement alternative pathway to induce acute lung injury in mice by intravenous injection. Myeloperoxidase（ MPO） activity of lung homogenate,the cell count and the protein content of bronchoalveolar lavage fluid（ BALF） were measured. The concentration of IL-6,TNF-α,P-selectin and ICAM-1 in BALF and serum were determined by ELISA. The pathological change of lung tissue was observed by HE staining. The phosphorylation of NF-κB p65 in lung tissues was checked by immunohistochemistry. The effect of the transcriptional activity of NF-κB signal pathway in microvascular endothelial cells was measured by employing dual-luciferase reporter assay system. Results ICA reduced MPO activity of lung homogenate,the cell count and the content of IL-6,TNF-α,P-selectin in BALF obviously.The level of TNF-α,P-selectin and ICAM-1 in serum was decreased,the pulmonary inflammatory cell infiltration was reduced,the phosphorylation of NF-κB p65 in lung was inhibited significantly and the transcriptional activity of NF-κB was also down-regulated. Conclusion ICA can alleviate acute inflammatory response of ALI mice induced by activation of the complement alternative pathway. The mechanism may be highly related to the inhibition of inflammatory cell infiltration in lung tissue,the down-regulation of phosphorylation of NF-κB p65 and nuclear transcriptional activity.

关 键 词：淫羊藿苷 补体旁路 急性肺损伤 眼镜蛇毒因子 NF-ΚB P65 炎症 

分 类 号：R-33[医药卫生] R284.1