日本血吸虫感染小鼠肝脏炎症反应与肠道菌群谱的相关性研究  

Correlation between liver inflammation and intestinal flora in mice infected with Schistosoma japonicum

作　　者：梅丛进 杨莹莹[1] 董盼盼 宋丽君[1] 周永华[1] 许永良[1] 余传信[1] MEI Congjin;YANG Yingying;DONG Panpan;SONG Lijun;ZHOU Yonghua;XU Yongliang;YU Chuanxin(Jiangsu Institute of Parasitic Diseases,National Health Commission Key Laboratory of Parasitic Disease Control and Prevention,Wuxi214064,Jiangsu,China)

机构地区：[1]江苏省血吸虫病防治研究所,国家卫生健康委员会寄生虫病预防与控制技术重点实验室,江苏无锡214064

出　　处：《中国病原生物学杂志》2023年第7期793-798,共6页Journal of Pathogen Biology

基　　金：无锡市科技发展基金项目(No.N20202040);无锡市卫生健康委科研项目(No.Q201937,Q202001);江南大学公共卫生研究中心青年项目(No.JHPU201834);所级人才项目(No.JHPU-RC-202005)。

摘　　要：目的观察和分析C57BL/6J小鼠感染日本血吸虫后肝脏病变与肠道菌群变化的相关性。方法将10只6周龄的C57BL/6J小鼠随机分为健康对照组和日本血吸虫感染组。感染组每只小鼠感染(15(1)条日本血吸虫尾蚴。在感染后的第7周解剖小鼠,取肝脏组织,1)经石蜡包埋后进行HE和Masson染色检查;2)采用比色法测定羟脯氨酸(HYP)含量;3)提取肝组织的mRNA,采用RT-PCR检测炎症因子IL-1b和TGF-b的mRNA水平;4)留取小鼠粪便,委托华大基因公司进行微生物多样性检测。结果小鼠肝脏切片经HE和Masson染色后检查,感染组平均单个肉芽肿和胶原面积在感染后7周分别为148532μm^(2)和193209μm^(2)。健康对照组和感染组羟脯氨酸含量分别为0.18和0.34μg/mg,差异有统计学意义(P<0.01)。RT-PCR检测健康对照组和感染组炎症因子IL-1b mRNA相对表达量分别为1.06和8.33,差异有统计学意义(P<0.01);纤维化因子TGF-bm RNA相对表达量分别为1.05和9.58,差异有统计学意义(P<0.01)。粪便16s多样性分析显示,感染组小鼠肠道菌群的Shannon指数低于健康对照组,Simpson指数高于健康对照组。门水平物种组成分析显示,血吸虫感染小鼠粪便中厚壁菌门(Firmicutes)丰度显著下调(P<0.05)。在属水平,感染小鼠粪便中拟杆菌属(Bacteroides),帕拉普菌(Paraprevotella),螺杆菌属(Helicobacter)的相对丰度均显著上调(均P<0.05),粪球菌属相对丰度显著下调(P<0.05)。对差异菌属与IL-1b、TGF-b和HYP进行环境因子关联分析,其中拟杆菌属、帕拉普菌和螺杆菌属丰度与IL-1b、TGF-b、HYP mRNA水平呈正相关,粪球菌属丰度与上述炎性因子mRNA相对水平呈负相关。结论小鼠感染日本血吸虫后肝脏虫卵肉芽肿样病变及炎症因子的mRNA相对表达水平变化与其肠道菌群组成的变化相关,拟杆菌属、帕拉普菌和螺杆菌属丰度与炎症因子及纤维化因子的mRNA水平呈正相关,粪球菌属丰度与上述炎性因�Objective The aim of this study is to observe and analyze the correlation between liver lesions and intestinal flora in C57BL/6J mice infected with Schistosoma japonicum.Methods 10 mice(6-week-old)were randomly divided into 2 groups,healthy control group and Schistosoma japonicum-infected group.Each mouse in the infected group was infected with 15 Schistosoma japonicum caecal larvae.The liver tissues were dissected at the 7th week after infection to perform the following analysis:1)After paraffin embedding,HE and Masson staining were performed;2)The content of hydroxyproline(HYP)was determined by colorimetry;3)The mRNA was extracted from the liver tissues,and the expression levels of inflammatory factors IL-1b and TGF-b in the liver tissues were analyzed by RT-PCR;4)The feces of mice were collected and tested for microbial diversity by BGI.Tech.Results After HE and Masson staining,the average area of single granuloma and collagen in the infected group was 148532μm^(2)and 193209μm^(2),respectively.The content of hydroxyproline in healthy control group and infection group was 0.18 and 0.34μg/mg,respectively,and the difference was statistically significant(P<0.01).The mRNA expression levels of IL-1b mRNA in healthy control group and infection group were 1.06 and 8.33,respectively,and the differences were statistically significant(P<0.01);the mRNA expression levels of fibrosis factor TGF-b were 1.05 and 9.58,respectively,and the difference was statistically significant(P<0.01).16s feces diversity analysis showed that the Shannon index of intestinal flora in infected group was lower than that in healthy control group,and Simpson index was higher than that in healthy control group.Analysis of phylum level species composition showed that the abundance of Firmicutes in infected mice feces was significantly decreased(P>0.05).At the genus level,the relative abundance of Bacteroides,Paraprevotella and Helicobacter in infected mice feces was significantly up-regulated(P>0.05),the relative abundance of Coprococcus was sign

关 键 词：日本血吸虫 虫卵肉芽肿炎症 肠道菌群 相互关系 

分 类 号：R383.24[医药卫生—医学寄生虫学]