青蒿琥酯对小鼠巨细胞病毒性肺炎的治疗作用  

Therapeutic effects of artesunate on cytomegalovirus pneumonia in mice

作　　者：王萍 陈盛 黄小桃 肖小梅 湛青平 秦爱萍 余细勇 WANG Ping;CHEN Sheng;HUANG Xiao-tao;XIAO Xiao-mei;ZHAN Qing-ping;QIN Ai-ping;YU Xi-yong(Guangdong Key Laboratory of Molecular Target and Clinical Pharmacology,State Key Laboratory of Respiratory Diseases Pharmacology Group,School of Pharmaceutical Sciences of Guangzhou Medical University,Guangzhou 511436,China)

机构地区：[1]广州医科大学药学院,广东省分子靶标与临床药理学重点实验室,呼吸疾病国家重点实验室药理学组,广东广州511436

出　　处：《药学学报》2020年第11期2651-2656,共6页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金重点项目(U1601227,81330007);广东省重大科技计划资助项目(GD2015B020225006)。

摘　　要：为了探索青蒿琥酯对小鼠巨细胞病毒性肺炎的治疗作用及其机制,应用滴鼻法建立携带绿色荧光蛋白的小鼠巨细胞病毒(murine cytomegalovirus-green fluorescent protein,MCMV-GFP)感染的巨细胞病毒性肺炎模型。动物福利和实验过程均遵循广州医科大学动物伦理委员会的规定。将BALB/c-nu小鼠随机分为5组:空白对照组、巨细胞病毒肺炎模型组、青蒿琥酯60、120和240 mg·kg^-1组。观察小鼠生存期和肺组织病理变化,并进一步进行分子机制研究。分别采用苏木精-伊红(hematoxylin-eosin,HE)染色法检测肺组织的病理变化;荧光定量PCR(polymerase chain reaction)方法检测肺组织中MCMV主要即刻早期基因1(major immediate early 1,Mie1)mRNA的表达;免疫荧光法检测MCMV-GFP的表达;ELISA法(enzyme-linked immunosorbent assay)检测肺上清炎症因子白细胞介素6(interleukin 6,IL-6)、IL-10和肿瘤坏死因子α(tumor necrosis factorα,TNF-α)的含量;Western blot法检测感染后肺组织NF-κB(nuclear factor-κB)通路的变化;绘制小鼠体重变化和生存曲线图。结果显示,MCMV感染模型组小鼠肺组织出现大量炎症细胞浸润,肺组织结构破坏。青蒿琥酯(120 mg·kg^-1)治疗后的小鼠肺部炎症减轻,肺泡结构明显好转。与MCMV感染模型组相比,青蒿琥酯(120和240 mg·kg^-1)剂量组治疗后,小鼠肺部病毒复制水平降低,青蒿琥酯(120 mg·kg^-1)剂量组可显著降低促炎因子IL-6和TNF-α水平并增加抗炎因子IL-10的水平;磷酸化NF-κB蛋白表达明显下降,治疗组小鼠体重增加,生存期延长。上述结果表明,青蒿琥酯具有抑制MCMV感染小鼠肺炎的作用,通过抑制NF-κB炎症信号通路的激活,减轻肺组织炎症因子IL-6和TNF-α的释放,并增加抗炎因子IL-10的表达,从而实现减轻肺部组织炎症的作用。To investigate the therapeutic effect of artesunate on mouse cytomegalovirus pneumonia,the BALB/c-nu mice were infected with murine cytomegalovirus-green fluorescent protein(MCMV-GFP)by nose dropping method.The experimental protocol was approved by the Medical Laboratory Animal Ethics Committee of Guangzhou Medical University.The BALB/c-nu mice were randomly divided into five groups:control group,MCMV pneumonia group,and artesunate(60,120,and 240 mg·kg^-1)groups.The survival rate,weights,and virus loads in lungs among the groups were observed.The degree of histopathologic changes in lungs was assessed directly by hematoxylin-eosin(HE)assay.MCMV-GFP expression was assessed by immunofluorescence.In addition,reverse transcription polymerase chain reaction(RT-PCR)analysis was performed to investigate the content of major immediate early 1(Mie1)mRNA,and enzyme-linked immunosorbent assay(ELISA)was used to detect the changes of inflammatory factors,interleukin 10(IL-10),IL-6,and tumor necrosis factor-α(TNF-α).Western blot analysis was used to detect the expression of the changes of nuclear factor-kappa B(NF-κB)signaling pathways in total proteins.Compared with MCMV group,artesunate(120 mg·kg^-1)significantly increased body weights of MCMV-infected nude mice over 30 days,and decreased the viral titer in lung homogenate,lung inflammation,and histological severity.Moreover,the administration of artesunate(120 mg·kg^-1)could downregulate the expression of phospho-NF-κB(p-NF-κB)p65 in the lungs of mice.The present study suggested that artesunate can protect the immunocompromised mice from MCMV-induced interstitial pneumonia via downregulating NF-κB signaling pathway,thus attenuating inflammation in the lungs.

关 键 词：巨细胞病毒 青蒿琥酯 肺炎 炎症因子 信号通路 

分 类 号：R965[医药卫生—药理学]