马拉龙和阿托伐醌+阿奇霉素在不同免疫状态小鼠体内的抗田鼠巴贝虫药效评价  

Evaluation on the in vivo efficacy of malarone and atovaquone-azithromycin combination against Babesia microti in mice under different immune status

作　　者：殷梦 张皓冰 陶奕 姜斌[1] 刘华[1] YIN Meng;ZHANG Hao-bing;TAO Yi;JIANG Bin;LIU Hua(National Institute of Parasitic Diseases,Chinese Center for Disease Control and Prevention,Chinese Center for Tropical Diseases Research,Key Laboratory on Parasite and Vector Biology,Ministry of Health,WHO Centre for Tropical Diseases,National Center for International Research on Tropical Diseases,Ministry of Science and Technology,School of Global Health,Chinese Center for Tropical Diseases,Jiaotong University School of Medicine,Shanghai 200025,China)

机构地区：[1]中国疾病预防控制中心寄生虫病预防控制所(国家热带病研究中心),卫生部寄生虫病原与媒介生物学重点实验室,世界卫生组织疟疾、血吸虫病和丝虫病热带病合作中心,科技部国家级热带病国际研究中心,上海交通大学医学院-国家热带病研究中心全球健康学院,上海200025

出　　处：《中国寄生虫学与寄生虫病杂志》2021年第5期659-665,673,共8页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：上海市公共卫生体系建设三年行动计划(2020-2022年)重点学科建设计划项目(GWV-10.1-XK13)。

摘　　要：目的以免疫状态正常的BALB/c小鼠及非肥胖糖尿病/重症联合免疫缺陷NOD/SCID小鼠为模型,对临床较常用的阿托伐醌(ATQ)+阿奇霉素(AZM)和马拉龙进行抗田鼠巴贝虫体内药效评价。方法取69只BALB/c健康小鼠与15只NOD/SCID健康小鼠,每鼠接种107个感染田鼠巴贝虫的鼠红细胞。2种小鼠感染后均设置3个组,即ATQ+AZM组(195 mg/kg ATQ+32.5 mg/kg AZM)、马拉龙组(62.5 mg/kg ATQ+25 mg/kg氯胍,即1/4片)和对照组(5%可溶性淀粉溶液),每鼠按0.2 ml/10 g灌胃给药。BALB/c小鼠药物抑制试验中(2个用药组各12只,对照组15只),小鼠感染后4 h开始用药,连用10 d,每组于感染后1、 3、 5、 7、 10 d喂药前,随机取3只小鼠尾尖采血,采用薄血膜涂片染色镜检观察红细胞感染情况并计算红细胞染虫率(EIR),qPCR检测18S r RNA基因相对量。BALB/c小鼠药物治疗试验中(3组各10只),于感染后7 d取所有小鼠尾尖血制薄血膜染色镜检,确认感染后开始用药,连用10 d,于感染后7、 10、 11、 12、 13、 15、 17、 19、 24、 27 d均采血镜检并计算EIR;于感染后27 d,每组随机取5只小鼠,采用免疫抑制剂地塞米松磷酸钠注射液(200μl/鼠),连续腹腔注射7 d,自免疫抑制后3 d起,每天采血制薄血膜涂片染色镜检,观察复燃情况;于感染后27 d,每组随机取5只小鼠采眶窦血,将抗凝全血混匀后腹腔注射接种对应的3组健康BALB/c小鼠(每组5只),继代接种感染后7~10 d,制薄血膜涂片染色镜检并计算EIR。NOD/SCID小鼠(3组各5只)于感染后10 d开始用药,连用10 d,于感染后10、 12、 15、 17、 19、 21、 24、 27、 29、 31、 35、 42、 49 d,分别取3组小鼠尾尖血,采用薄血膜涂片染色镜检并计算EIR。应用GraphPad Prism 8对数据进行统计学分析。结果 BALB/c小鼠药物抑制试验结果显示,ATQ+AZM及马拉龙均可有效抑制小鼠虫血症。镜检结果显示,ATQ+AZM组在感染后3 d、5 d,均仅1只小鼠查见虫体,EIR分别为(0.20�Objective To evaluate the in vivo efficacy against Babesia microti of two therapies commonly used in the clinic: a combination of atovaquone(ATQ) with azithromycin(AZM) and malarone in immune-normal BALB/c and non-obese diabetic/severe combined immunodeficient(NOD/SCID) mice models. Methods In total, 69 BALB/c and 15 NOD/SCID mice were each inoculated with 107 Babesia microti-infected erythrocytes. The mice of each immune type were divided into three groups: ATQ + AZM group(195 mg/kg ATQ + 32.5 mg/kg AZM), malarone group(62.5 mg/kg ATQ + 25 mg/kg proguanil), and control group(5% soluble starch solution), the drug was administered by gavage of 0.2 ml/10 g body weight. In the drug-suppression testing in BALB/c mice(12 mice each of two drug groups, and 15 mice in the control group), the mice received the first dose of drug 4 h after infection,and then the dosing was continued for 10 consecutive days. Three mice were randomly selected for blood sampling from tail tip before gavage administration on 1, 3, 5, 7, and 10 days post-infection. Thin blood smears were prepared for microscopy to examine B. microti infection of erythrocytes, and estimate the erythrocyte infection rate(EIR);the blood samples were tested for 18 S r RNA using qPCR to examine the gene value. In the drug-therapy testing experiment with BALB/c mice(10 mice each of 3 group), tail tip blood samples were collected from all mice 7 d post-infection to determine whether the infection was established, then when the infection was confirmed,the drug administration was initiated and continued for 10 consecutive days;microscopic examination was conducted for the blood samples collected on days 10, 11, 12, 13, 15, 17, 19, 24 and 27 post-infection to estimate the EIR.On 27 d post-infection, 5 mice randomly selected from each group were intraperitoneally injected with immunosuppressant dexamethasone sodium phosphate solution(200 μl/mouse) for 7 consecutive days;starting on 3 d after immunosuppressant injection, daily blood samples were examined by staining micro

关 键 词：田鼠巴贝虫 阿托伐醌 阿奇霉素 马拉龙 药效评价 红细胞染虫率 

分 类 号：R531.9[医药卫生—内科学]