环鸟苷酸腺苷酸促日本血吸虫感染小鼠肝虫卵肉芽肿形成及纤维化  

Cyclic guanosine monophosphate-adenosine monophosphate promotes liver egg granuloma formation and fibrosis in mice infected with Schistosoma japonicum

作　　者：梁乐 张璟[1] 沈玉娟 胡媛 曹建平 LIANG Le;ZHANG Jing;SHEN Yu-juan;HU Yuan;CAO Jian-ping(National Institute of Parasitic Diseases,Chinese Center for Disease Control and Prevention(Chinese Center for Tropical Diseases Research),NHC Key Laboratory of Parasite and Vector Biology,WHO Collaborating Center for Tropical Diseases,National Center for International Research on Tropical Diseases,Shanghai 200025,China;Shanghai University of Medicine&Health Sciences,Shanghai 201328,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),国家卫生健康委寄生虫病原与媒介生物学重点实验室,世界卫生组织热带病合作中心,国家级热带病国际联合研究中心,上海200025 [2]上海健康医学院,上海201328

出　　处：《中国寄生虫学与寄生虫病杂志》2022年第4期441-445,453,共6页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家自然科学基金面上项目(81971969,81772225);上海市公共卫生体系建设三年行动计划(2020—2022)重点学科项目(GWV-10.1-XK13)。

摘　　要：目的研究第二信使分子环鸟苷酸腺苷酸(cGAMP)对日本血吸虫感染小鼠肝虫卵肉芽肿形成及纤维化的影响,并初步阐明其调控机制。方法将C57BL/6小鼠随机分为cGAMP处理组(8只)、感染对照组(28只)、感染cGAMP处理组(28只)。cGAMP处理组尾静脉单独注射cGAMP(2μg,100μl),感染对照组和感染cGAMP处理组经腹部皮肤贴片感染日本血吸虫尾蚴[(20±2)条/鼠]后,分别经尾静脉注射PBS(100μl)和cGAMP(2μg,100μl),每周1次。3组小鼠各8只分别注射10次,每次注射后观察小鼠死亡情况。感染对照组和感染cGAMP处理组小鼠(各5只)于感染后7周,取小鼠眼眶血,ELISA检测丙氨酸氨基转移酶(ALT)与天冬氨酸氨基转移酶(AST)的水平;取小鼠肝组织切片进行苏木精-伊红染色(HE)和Masson染色,计算单个虫卵肉芽肿面积和肝纤维化面积,并计数肝组织虫卵数。感染对照组和感染cGAMP处理组小鼠(各15只)于感染后0、4和7周,取小鼠肝组织,提取总RNA,实时荧光定量PCR(qRT-PCR)检测肝组织β干扰素(IFN-β)编码基因Ifnb1的表达水平;取小鼠眼眶血,ELISA检测血清中IFN-β水平。采用GraphPad Prism 8.0.2软件进行统计学分析,两组间比较采用t检验。结果cGAMP处理组小鼠无死亡;感染后10周,感染对照组小鼠死亡2只,感染cGAMP处理组小鼠死亡5只,两组差异有统计学意义(χ^(2)=7.55,P<0.01)。感染对照组和感染cGAMP处理组小鼠血清ALT分别为(394.80±72.82)和(627.20±93.58)U/L(t=4.38,P<0.01),血清AST分别为(605.00±148.90)和(871.80±138.10)U/L(t=2.94,P<0.05)。感染对照组和感染cGAMP处理组小鼠肝组织虫卵数分别为50600±15473和53400±16273,差异无统计学意义(t=0.28,P>0.05);HE染色结果显示,感染对照组和感染cGAMP处理组均出现明显的肝组织结构被破坏,并有炎性细胞浸润,肝组织病理损害比例分别为(35.32±15.09)%和(64.98±13.91)%(t=3.23,P<0.05);肝组织虫卵肉芽肿面积分别为(0.57±0.45)和(1.17±0.74)mm^(2)Objective To investigate the effects of cyclic guanosine monophosphate-adenosine monophosphate(cGAMP),a second messenger molecule,on liver egg granulomas and fibrosis in Schistosoma japonicum-infected mice and to preliminarily clarify its regulatory mechanism.Methods C57BL/6 mice were randomly divided into cGAMP-treated group(8 mice),infected control group(28 mice),and infected cGAMP-treated group(28 mice).For the cGAMP-treated group,cGAMP(2μg,100μl)was injected via the tail vein,while the infected control group and the infected cGAMP-treated group were injected with PBS(100μl)and cGAMP(2μg,100μl)via the tail vein once a week,respectively,after being infected with S.japonicum cercariae[(20±2)/mouse]via abdominal skin patch.From each group,8 mice received 10 ingections to observe survivl state after each injection.After 7 weeks,mice orbital blood samples from infected control group and infected cGAMP-treated group(each 5)were collected to detect the levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)by ELISA.The liver tissues sections were prepared and stained with hematoxylin eosin(HE)and Masson to estimate the area of individual egg granuloma and fibrosis,and count the number of eggs in the liver tissues.Mice liver tissues were collected from infected control group and infected cGAMP-treated group(15 each),on 0,4,and 7 weeks post-infection,for extraction of total RNA,which was used for performing real time fluorescent quantitative PCR(qRT-PCR)to examine the expression level of IFN-β-encoding gene Ifnb1;the mice orbital blood was collected to detect the serumβ-interferon(IFN-β)level by ELISA.GraphPad Prism 8.0.2 software was used for statistical analysis,and t-test was used for comparison between the two groups.Results There were no deaths in the cGAMP-treated group,and 10 weeks after infection,2 mice died in the infected control group and 5 mice died in the infected cGAMP-treated group,with statistically significant differences between the two groups(χ^(2)=7.55,P<0.01).The number

关 键 词：日本血吸虫 环鸟苷酸腺苷酸 干扰素基因刺激蛋白 Ⅰ型干扰素 

分 类 号：R383.24[医药卫生—医学寄生虫学]