穿心莲内酯通过抗炎和调节免疫提高EV71感染小鼠的生存率  

Andrographolide protects mice against severe Enterovirus 71 infection by its anti-inflammation and immunomodulation effects

作　　者：管晨[1] 李敏[2] 任庆杰[3] 张伟[4] 单敏敏[1] 王融冰[4] 张广州[2] 刘清泉[5] 王希良[3] 赵忠鹏[3] 王玉光[5] 

机构地区：[1]北京中医药大学第一临床学院中医内科,100029 [2]军事医学科学院实验动物中心,北京100071 [3]军事医学科学院微生物流行病研究所免疫学研究室病原微生物生物安全国家重点实验室,北京100071 [4]首都医科大学附属北京地坛医院感染科,100015 [5]首都医科大学附属北京中医医院呼吸科,100010

出　　处：《免疫学杂志》2013年第9期737-744,共8页Immunological Journal

基　　金：“重大新药创制”科技重大专项(2011ZX09201-201-04);国家自然科学基金(30901204);北京市科技基金(JJT-2010-18)

摘　　要：目的阐明穿心莲内酯磺化物(喜炎平注射液)防治手足口病(hand,foot and mouth disease,HFMD)的效果和机制。方法将不同质量浓度的喜炎平注射液分别作用于Vero细胞和7日龄乳鼠,观测细胞的CPE和乳鼠的死亡率,测定喜炎平注射液的CC50(50%cell cytotoxicity)和MLD(median lethal dose);将1.25×107TCID50的71型肠道病毒(Enterovirus 71,EV71)BJ09/07毒株腹腔注射7日龄ICR乳鼠,建立HFMD重症感染模型;在喜炎平注射液无毒浓度下,通过比较喜炎平注射液药物组,相对于生理盐水对照组对EV71重症感染模型临床体征改善情况,测定喜炎平注射液防治EV71感染的效果;通过比较药物组,相对于对照组对EV71感染Vero细胞CPE的改善情况,测定喜炎平注射液的抗病毒效果;通过比较药物组和对照组的组织病变程度、炎症因子浓度、脾淋巴细胞增殖和激活程度,初步探索喜炎平注射液防治EV71的细胞和分子机制;通过比较药物组和对照组中性粒细胞MAPK/NF-κB信号通路分子表达和激活状态,初步揭示喜炎平注射液防治EV71的信号传导机制。结果在无毒质量浓度条件下,喜炎平注射液可显著降低EV71重症感染模型的死亡率和临床疾病得分,降低组织病变程度和炎症分子的质量浓度,提升脾T淋巴细胞的数量和激活状态;而喜炎平注射液防治HFMD的效果不是通过抗EV71病毒作用,而是主要通过降低炎性信号通路信号分子的表达和激活,从而抑制炎症因子分泌、改善机体细胞免疫状态,进而减轻组织炎症损伤而获得。结论喜炎平注射液可通过调节免疫治疗EV71感染引起的重症HFMD,暗示穿心莲内酯衍生物可能是1种有效的HFMD防治药物。Hand,foot and mouth disease（HFMD） which caused by Enterovirus 71（EV71） in Asia broke out in recent years.Andrographolide derivatives are widely used in treating HFMD in China,but the benefits and harms of those uses in HFMD have not been clearly elucidated.In this study,we aimed to investigate the efficacy and mechanism of andrographolide（Xiyanping） against HFMD.Firstly,seven-day-old suckling ICR mice were infected with 1.25×107 TCID50 EV71 BJ09/07 strain to prepare HFMD mouse models.And then,Xiyanping injection was given to mice intraperitoneally post infection.Routine clinical parameters and histopathologic changes were examined.In addition,the molecular mechanism of Xiyanping against HFMD was explored by analyzing the antiviral,anti-inflammatory and immunomodulatory effects and testing the MAPKs/NF-κB signaling pathway.All the quantitative data were performed by one way ANOVA analysis following Student＇s t-test.Treatment with Xiyanping injection could result in reduction of mortality,clinical scores and pathological changes which were possibly achieved through the anti-inflammatory and immunomodulatory activities of Xiyanping.The serum and muscle IL-6,TNF-alpha and MCP-1 levels were significantly decreased in HFMD mice treated with Xiyanping injection,compared with Saline-treated HFMD mice in the same conditions.In addition,markedly elevated splenic activated T immune cells were observed in the protected mice.Furthermore,there was no significant difference in virus titers between Xiyanping injection-treated mice and Saline-treated ones,indicating that the Xiyanping injection-mediated protection is independent of the virus load.According to pathway analysis,Xiyanping injection significantly inhibited NF-κB expression and down-regulated the levels of p-ERK and p-JNK in EV71 infected neutrophils.Taken together,andrographolide derivatives may be an effective medicine for the clinical treatment of HFMD patients.

关 键 词：肠道病毒71 穿心莲内酯衍生物 抗炎 免疫调节 

分 类 号：R285.5[医药卫生—中药学]