广东农业科学2025，Vol.52Issue(7)：72-83,12.DOI:10.16768/j.issn.1004-874X.2025.07.007
基于16S rRNA基因测序探究ApoE基因敲除兔肠道微生物特征
Exploring the Intestinal Microbiota Characteristics in ApoE Knockout Rabbits Based on 16S rRNA Gene Sequencing
摘要
Abstract
[Objective]To investigate the characteristics and differences of gut microbial communities between ApoE knockout rabbits and New Zealand rabbits,and to elucidate the potential role of the ApoE gene in regulating intestinal microbiota.[Method]ApoE knockout rabbits were successfully generated using CRISPR/Cas9 technology.High throughput sequencing of 16S rRNA gene sequences from fecal samples of ApoE knockout rabbits and New Zealand rabbits was performed on the NovaSeq sequencing platform.Bioinformatics analyses-including quality control,ASV(amplicon sequence variant)clustering,taxonomic annotation,and diversity assessments were conducted to systematically evaluate the differences in gut microbiota diversity,community structure,and functional regulation.[Result]Under normal dietary conditions,ApoE knockout rabbits exhibited higher levels of total cholesterol(TC),triglycerides(TG),and low-density lipoprotein cholesterol(LDL-C)in serum compared to New Zealand rabbits,with differences in TC and LDL-C being highly significant(P<0.01),consistent with hyperlipidemia characteristics.Compared with New Zealand rabbits,ApoE knockout rabbits showed a significant increase in gut microbial ASV abundance(P<0.01);α diversity indices including Observed_species,Chao1,Shannon,Simpson,and PD_whole_tree were significantly elevated(P<0.05),indicating increased species richness and diversity.β diversity analysis revealed significant differences,with principal component and principal coordinate analyses explaining 28.84%and 38.20%of the variance respectively,suggesting marked alterations in gut microbial community structure.The Firmicutes/Bacteroidetes ratio(F/B)was significantly increased in ApoE knockout rabbits(P<0.05),the gut shows an increase in lipopolysaccharide(LPS)-producing bacteria and mucosa-damaging bacteria such as Akkermansia,Alistipes,and Clostridium.Additionally,significant differences were observed in pathways related to basal transcription factor regulation,drug metabolism,and atrazine degradation(P<0.05).[Conclusion]The gut microbial diversity and metabolic pathways in ApoE gene knockout rabbits were altered,indicating that the deletion of the ApoE gene affects the gut microbial ecology and metabolic functions,playing a key role in regulating intestinal microecological balance.关键词
兔/高脂血症/ApoE基因敲除/CRISPR/Cas9/16S rRNA/肠道微生物Key words
rabbits/hyperlipidemia/ApoE knockout/CRISPR/Cas9/16S rRNA/intestinal microbiota分类
农业科技引用本文复制引用
黎桂玲,萨楚拉,陈傍柱,刘月姝,刘科,谭巧燕,郑佳琳,朱叶萌,王刚..基于16S rRNA基因测序探究ApoE基因敲除兔肠道微生物特征[J].广东农业科学,2025,52(7):72-83,12.基金项目
广东省科技计划项目(2019A030317012) （2019A030317012）
广东省医学科研基金项目(B2024060) （B2024060）
