阻断白细胞介素9对日本血吸虫感染小鼠免疫病理损伤及肠道菌群的影响  

Effects of interleukin-9 blockage on immunopathological injury and gut microbiota during Schistosoma japonicum infection in mice

作　　者：吕嘉慧 王纪龙 肖苏羽 卢作超[1] 莫景筌 李彩淇 邱然 唐泽丽 李青 战廷正[1] LYU Jiahui;WANG Jilong;XIAO Suyu;LU Zuochao;MO Jingquan;LI Caiqi;QIU Ran;TANG Zeli;LI Qing;ZHAN Tingzheng(Department of Parasitology,Guangxi Medical University,Nanning,Guangxi 530021,China;School of Pre-clinical Medicine,Guangxi Medical University,Nanning,Guangxi 530021,China;Department of Cell Biology and Genetics,Guangxi Medical University,Nanning,Guangxi 530021,China)

机构地区：[1]广西医科大学寄生虫学教研室,广西南宁530021 [2]广西医科大学临床医学院,广西南宁530021 [3]广西医科大学细胞生物学教研室,广西南宁530021

出　　处：《热带医学杂志》2024年第5期611-616,F0002,共7页Journal of Tropical Medicine

基　　金：国家自然科学基金(82260407,81860358);广西壮族自治区自然科学基金(2018GXNSFAA281059,2023GXNSFBA026068);国家级大学生创新创业训练计划项目(202210598014)

摘　　要：目的研究阻断白细胞介素9(IL-9)对日本血吸虫(Sj)感染小鼠肝组织免疫损伤以及肠道菌群的影响,探讨阻断IL-9改善血吸虫病免疫病理损伤的机制。方法将15只实验小鼠随机分成正常对照组、Sj组和IL-9阻断组。Sj组和IL-9阻断组小鼠分别感染(15±1)条日本血吸虫尾蚴,IL-9阻断组小鼠每周腹腔注射IL-9抗体2次,Sj组小鼠每周腹腔注射IgG抗体2次作为同型对照。感染9周后处死小鼠,收取肝脏组织进行苏木精-伊红(HE)染色、Masson染色及免疫组化;收取小鼠结肠内容物,对肠道微生物进行宏基因组测序。结果HE、Masson染色显示,单个肉芽肿面积百分比,Sj组(58.6±4.9)%>IL-9阻断组(37.0±2.1)%,差异有统计学意义(F=678.4,P<0.001)。胶原沉积面积百分比,Sj组(35.5±2.0)%>IL-9阻断组(16.2±0.3)%>正常对照组(0.4±0.1)%,差异有统计学意义(F=1314.0,P<0.001)。免疫组化染色显示,IL-9阻断后小鼠肝组织α-平滑肌动蛋白(α-SMA)、胶原Ⅰ和胶原Ⅲ蛋白的表达水平显著降低。α-SMA蛋白表达百分比,Sj组(29.9±4.0)%>IL-9阻断组(14.9±1.4)%>正常对照组(1.4±0.4)%,差异有统计学意义(F=267.9,P<0.001);胶原Ⅰ蛋白表达百分比,Sj组(33.9±1.2)%>IL-9阻断组(16.2±0.7)%>正常对照组(1.3±0.2)%,差异有统计学意义(F=3112.0,P<0.001);胶原Ⅲ蛋白表达百分比,Sj组(14.5±3.0)%>IL-9阻断组(5.3±0.6)%>正常对照组(1.4±0.2)%,差异有统计学意义(F=131.1,P<0.001)。小鼠肠道微生物宏基因组测序显示,与正常对照组比较,Sj组和IL-9阻断组小鼠肠道菌群的多样性和物种丰富度均降低,其中IL-9阻断组与正常对照组差异有统计学意义(P=0.002),温生菌、巴恩斯氏菌、未分类肠杆菌、埃希氏菌属等的丰度发生显著变化。结论体内阻断IL-9可改变日本血吸虫感染小鼠肠道菌群的物种结构、多样性,并且这种改变可能与日本血吸虫病免疫病理的发生发展密切相关。Objective To evaluate the effects of interleukin-9(IL-9)blockage on liver tissue immune damage and gut microbiota in mice infected with Schistosoma japonicum(Sj),and explore the mechanism of IL-9 blockage improving immunopathological injury in schistosomiasis.Methods Fifteen mice were randomly divide into control group,Sj group,and IL-9 blockage group.Each mouse in the Sj group and the IL-9 blockage group was infected with(15±1)S.japonicum cercariae.Each mouse in the IL-9 blockage group was intraperitoneally injected with anti-IL-9 antibody twice a week.Mice in the Sj group were injected with Ig G antibody twice a week as an isotype control.At week 9 post-infection,mice were sacrificed and their liver tissues were examined by hematoxylin-eosin(HE)staining,Masson staining and immunohistochemistry.Colon contents were collected from mice and perform metagenomic sequencing on gut microbiota.Results HE and Masson staining showed that the percentage of individual granuloma area,Sj group(58.6±4.9)%>IL-9 blockage group(37.0±2.1)%;the difference was statistically significant(F=678.4,P<0.001).Percentage of collagen deposition area,Sj group(35.5±2.0)%>IL-9 blockage group(16.2±0.3)%>normal control group(0.4±0.1)%;the difference was statistically significant(F=1314.0,P<0.001).Immunohistochemistry showed that IL-9 blockage markedly declined the expression levels ofα-smooth muscle actin(α-SMA),collagenⅠ,and collagenⅢproteins in liver tissue of mice.Percent ofα-SMA protein expression,the Sj group(29.9±4.0)%>IL-9 blockage group(14.9±1.4)%>normal control group(1.4±0.4)%;the differences was statistically significant(F=267.9,P<0.001).Percent of collagenⅠprotein expression,Sj group(33.9±1.2)%>IL-9 blockage group(16.2±0.7)%>normal control group(1.3±0.2)%;the difference was statistically significant(F=3112.0,P<0.001).Percentage of collagenⅢexpression,Sj group(14.5±3.0)%>IL-9 blockage group(5.3±0.6)%>normal control group(1.4±0.2)%;the difference was statistically significant(F=131.1,P<0.001).Metagenomic sequenc

关 键 词：日本血吸虫 虫卵肉芽肿 肝纤维化 白细胞介素9 肠道菌群 

分 类 号：R383.24[医药卫生—医学寄生虫学]