目的研究固醇调节元件结合蛋白1c(sterol regulatory element binding protein 1c,SREBP1c)缺失对小鼠肠道菌群结构的影响。方法以SREBP1c基因敲除鼠为研究对象,收集纯合敲除型(SREBP1c-KO)和同窝野生型(SREBP1c-WT)的小鼠粪便,采用16S ...目的研究固醇调节元件结合蛋白1c(sterol regulatory element binding protein 1c,SREBP1c)缺失对小鼠肠道菌群结构的影响。方法以SREBP1c基因敲除鼠为研究对象,收集纯合敲除型(SREBP1c-KO)和同窝野生型(SREBP1c-WT)的小鼠粪便,采用16S rDNA测序粪便基因组DNA,采用物种分类学分析、多样性指数分析、聚类分析、主成分分析(principal component analysis,PCA)等,比较敲除型和野生型两组小鼠肠道菌群结构的组成差异。检测两组小鼠肝脏脂合成代谢以及肠道屏障的差异。结果常规饲养下,SREBP1c基因敲除对小鼠的体重及肝脏总胆固醇含量(total cholesterol,TC)没有显著影响,明显减少肝脏内甘油三酯(triglyceride,TG)含量。α多样性指数(Sobs指数)分析发现,SREBP1c-KO小鼠的肠道菌群多样性显著低于野生型小鼠(P<0.01);PCA分析显示,两组小鼠肠道菌群组成有一定程度的差异。在门水平,厚壁菌门与拟杆菌门比值在SREBP1c-KO小鼠肠道中有上升趋势,但没有统计学差异;在属水平,相较于野生型小鼠,SREBP1c-KO小鼠肠道的理研菌属(Rikenella)相对丰度显著减少(P<0.05),同时肠道抗菌肽Reg3γ和紧密连接蛋白Occludin表达显著下调。结论脂代谢关键调控基因SREBP1c缺失降低小鼠肠道菌群多样性,减少理研菌属相对丰度水平,可能与肠道屏障减弱有关。展开更多
Objective To verify the neurotypicality of AAV-PHP.eB after tail vein injection in adult mice and its efficiency in crossing the blood-brain barrier(BBB).Methods The rAAV-SYN-GFP plasmid was constructed, and adult C57...Objective To verify the neurotypicality of AAV-PHP.eB after tail vein injection in adult mice and its efficiency in crossing the blood-brain barrier(BBB).Methods The rAAV-SYN-GFP plasmid was constructed, and adult C57 BL mice were injected with AAV PHP.eB: SYN-GFP in the tail vein(300 nL, virus titer 3 × 10~9 vg) and in the prefrontal lobe(50 L, virus tite5 × 10^(11) vg). The green fluorescent protein(GFP) signal in the brain was observed at two weeks, while the GFP signal in the peripheral organs was observed at four weeks. Results Two weeks after tail vein injection, GFP expression was observed throughout the brain especially in the cortex, hippocampus, and geniculate nucleus. No GFP signal was observed or detected by western blotting in the peripheral organs after four weeks. GFP signal was observed mainly at the loca site after prefrontal lobe injection.Conclusion AAV-PHP.eB: SYN-GFP can effectively cross the BBB in adult mice. Using a neuron-specific promoter allows exogenous gene expression in neurons; therefore, AAV-PHP.eB can be used as an effective carrier for studying diseases in the central nervous system(CNS).展开更多
文摘目的研究固醇调节元件结合蛋白1c(sterol regulatory element binding protein 1c,SREBP1c)缺失对小鼠肠道菌群结构的影响。方法以SREBP1c基因敲除鼠为研究对象,收集纯合敲除型(SREBP1c-KO)和同窝野生型(SREBP1c-WT)的小鼠粪便,采用16S rDNA测序粪便基因组DNA,采用物种分类学分析、多样性指数分析、聚类分析、主成分分析(principal component analysis,PCA)等,比较敲除型和野生型两组小鼠肠道菌群结构的组成差异。检测两组小鼠肝脏脂合成代谢以及肠道屏障的差异。结果常规饲养下,SREBP1c基因敲除对小鼠的体重及肝脏总胆固醇含量(total cholesterol,TC)没有显著影响,明显减少肝脏内甘油三酯(triglyceride,TG)含量。α多样性指数(Sobs指数)分析发现,SREBP1c-KO小鼠的肠道菌群多样性显著低于野生型小鼠(P<0.01);PCA分析显示,两组小鼠肠道菌群组成有一定程度的差异。在门水平,厚壁菌门与拟杆菌门比值在SREBP1c-KO小鼠肠道中有上升趋势,但没有统计学差异;在属水平,相较于野生型小鼠,SREBP1c-KO小鼠肠道的理研菌属(Rikenella)相对丰度显著减少(P<0.05),同时肠道抗菌肽Reg3γ和紧密连接蛋白Occludin表达显著下调。结论脂代谢关键调控基因SREBP1c缺失降低小鼠肠道菌群多样性,减少理研菌属相对丰度水平,可能与肠道屏障减弱有关。
基金Supported by grants from the Innovation of Science and Technology Talents in Harbin(No.2017RAXQJ045)the Fundamental Research Funds for the Central Universities
文摘Objective To verify the neurotypicality of AAV-PHP.eB after tail vein injection in adult mice and its efficiency in crossing the blood-brain barrier(BBB).Methods The rAAV-SYN-GFP plasmid was constructed, and adult C57 BL mice were injected with AAV PHP.eB: SYN-GFP in the tail vein(300 nL, virus titer 3 × 10~9 vg) and in the prefrontal lobe(50 L, virus tite5 × 10^(11) vg). The green fluorescent protein(GFP) signal in the brain was observed at two weeks, while the GFP signal in the peripheral organs was observed at four weeks. Results Two weeks after tail vein injection, GFP expression was observed throughout the brain especially in the cortex, hippocampus, and geniculate nucleus. No GFP signal was observed or detected by western blotting in the peripheral organs after four weeks. GFP signal was observed mainly at the loca site after prefrontal lobe injection.Conclusion AAV-PHP.eB: SYN-GFP can effectively cross the BBB in adult mice. Using a neuron-specific promoter allows exogenous gene expression in neurons; therefore, AAV-PHP.eB can be used as an effective carrier for studying diseases in the central nervous system(CNS).
