基于16S rDNA高通量测序探讨肠道菌群对疟原虫感染小鼠的保护性作用  

Investigation of protective research of gut microbiome on Plasmodium infection mice by 16S rDNA high-throughput sequencing

作　　者：田爽 吴双 李欣 廖国燕 杜云婷 陈光[1,2] TIAN Shuang;WU Shuang;LI Xin;LIAO Guo-yan;DU Yun-ting;CHEN Guang(College of Basic Medicine,Jiamusi University,Jiamusi 154000,China;College of Medicine,Taizhou College,Taizhou 318000,China;Hebei North University,Zhangjiakou 075000,China;Department of Clinical Laboratory,Tumor Hospital of China Medical University,Liaoning Tumor Hospital,Shenyang 110042,China)

机构地区：[1]佳木斯大学基础医学院,佳木斯154000 [2]台州学院医学院,台州318000 [3]河北北方学院,张家口075000 [4]中国医科大学肿瘤医院,辽宁省肿瘤医院检验科,沈阳110042

出　　处：《中国人兽共患病学报》2023年第2期99-106,共8页Chinese Journal of Zoonoses

基　　金：国家自然科学基金(No.81101278);台州学院杰出青年项目(No.2019JQ003)。

摘　　要：目的探究Plasmodium berghei ANKA(P.bANKA)感染小鼠肠道菌群特异性改变及其保护性研究。方法C57BL/6小鼠随机分为正常对照组(NC组)和P.bANKA感染组(P.b A组),动态留取粪便进行16S rDNA高通量测序。建立益生菌预处理组(Pre-probiotics+P.b A组),伊文思蓝灌注评估血脑屏障(BBB)完整性,苏木精-伊红(HE)染色,观察脑组织病理学改变。结果16S rDNA高通量测序分析,P.bANKA感染小鼠后肠道菌群以乳酸杆菌和罗伊乳酸杆菌明显下降为主要变化趋势,且呈时间依赖性。动物验证实验发现,与P.b A组相比,Pre-probiotics+P.b A组原虫血症水平较低,小鼠于感染后第14 d开始出现死亡且未发生脑疟神经系统症状,BBB完整性较好,脑组织无明显损伤。结论P.bANKA感染可明显改变宿主肠道菌群的结构和功能,其中以乳酸杆菌数量减少最为明显。外源性补充乳酸杆菌后可明显降低ECM小鼠原虫血症水平,提高生存率。其作用机制可能与保护BBB完整性,改善小鼠脑组织病理损伤密切相关。The aim of this study was to investigate the specific changes and protective effects of the intestinal flora in mice infected with Plasmodium berghei ANKA(P.bANKA).C57BL/6 mice were randomly divided into a normal control group(NC group)and P.bANKA infection group(P.b A group),and feces were collected for 16S rDNA high-throughput sequencing.A pre-probiotic+P.b A group was established.Evans blue stain was perfused to assess blood-brain barrier(BBB)integrity,and hematoxylin-eosin staining was used to observe histopathological changes in brain tissue.In high-throughput sequencing analysis of 16S rDNA,the main trend in the intestinal flora in mice infected with P.bANKA indicated a significant time-dependent decrease in Lactobacillus and Lactobacillus royale.Animal validation experiments demonstrated lower parasitemia in the pre-probiotic+P.b A group than the P.b A group;the mice started to die 14 days after infection and did not develop neurological symptoms of cerebral malaria.The integrity of the BBB was better,and no obvious damage to brain tissue was observed.P.bANKA infection significantly altered the structure and function of the host intestinal flora,thus significantly decreasing the number of Lactobacillus.Exogenous supplementation with Lactobacillus significantly decreased parasitemia in ECM mice and increased the survival rate.The underlying mechanism may be closely related to protection of BBB integrity and amelioration of pathological damage to mouse brain tissue.

关 键 词：16S rDNA Plasmodium berghei ANKA 肠道菌群 高通量测序 脑疟 乳酸杆菌 

分 类 号：R446[医药卫生—诊断学]