摘要
目的基于内源转录间隔区(ITS)rRNA基因高通量测序技术探讨脾虚高脂血症大鼠肠道真菌多样性及构成的改变,以及香砂六君子汤对其的影响。方法选择15只6周龄体重180~200 g的SD大鼠依照随机数表法分为正常对照组(Z组)、脾虚高脂血症组(M组)、香砂六君子汤组(XS组),每组5只。M组大鼠采用高脂饲料+脾虚复合方法造模,共12周;XS组大鼠造模12周后(造模方法同上),灌胃香砂六君子汤4周,其余两组大鼠每日灌胃生理盐水4周。使用生化试剂盒检测血脂水平。采用油红O染色观察各组大鼠肝脏脂质沉积情况。应用PAS染色观察各组大鼠小肠杯状细胞变化。使用ITS rRNA基因高通量测序技术检测各组大鼠肠道真菌菌群多样性及构成变化。结果香砂六君子汤可以显著降低血清总胆固醇(F=14.861,P<0.01)、甘油三酯(F=13.377,P<0.01)水平,对低密度脂蛋白胆固醇(F=60.481,P>0.05)有降低趋势,显著升高高密度脂蛋白胆固醇(F=102.197,P<0.01)水平,缓解肝脏中大量脂质沉积现象,增加小肠内杯状细胞数量。肠道真菌检测结果显示Z组特有物种405个、M组特有物种238个、XS组特有物种201个;显著下降脾虚高脂血症组大鼠的Chao1指数(F=22.070,P<0.01)、ACE指数(F=29.280,P<0.01)、PD whole-tree指数(F=211.000,P<0.01)和Pielou指数(F=11.360,P<0.05),菌群丰度多样性降低,香砂六君子汤可以逆转大鼠肠道真菌丰度多样性;门水平上M组大鼠担子菌门/子囊菌门比值显著降低(F=4.824,P<0.05),给予香砂六君子汤后可进一步逆转,但无显著差异;属水平中百分占比较多的真菌属为丛赤壳属、枝孢属、多球壳属;菌群生态功能分析表明三组肠道真菌主要的营养方式为腐生。结论香砂六君子汤可能通过改变脾虚高脂血症大鼠肠道真菌丰度和多样性来改善脾虚高脂血症大鼠血脂代谢紊乱。
Objective To investigate the changes in intestinal fungal diversity and composition in rats with spleen deficiency hyperlipidemia and the intervention effect of Xiangsha Liujunzi Decoction based on high-throughput sequencing technology of internal transcribed spacer(ITS)rRNA gene.Methods A total of 156-week-old SD rats weighing 180-200 g were randomly divided into normal control group(Group Z),spleen deficiency hyperlipidemia group(Group M),and Xiangsha Liujunzi Decoction group(Group XS),with 5 rats in each group.rats in Group M were modeled by high-fat diet combined with spleen deficiency method for 12 weeks;after 12 weeks of modeling(using the same modeling method as above),rats in Group XS were given Xiangsha Liujunzi Decoction by gavage for 4 weeks,while rats in the other two groups were given normal saline by gavage daily for 4 weeks.Biochemical kits were used to detect blood lipid levels.Oil red O staining was used to observe the lipid deposition in the liver of rats in each group.PAS staining was applied to observe the changes of goblet cells in the small intestine of rats in each group.High-throughput sequencing technology of ITS rRNA gene was used to detect the diversity and composition changes of intestinal fungal flora in rats of each group.Results Xiangsha Liujunzi Decoction could significantly reduce serum total cholesterol(F=14.861,P<0.01)and triglycerides(F=13.377,P<0.01),had a decreasing trend on low-density lipoprotein cholesterol(F=60.481,P>0.05),significantly increase high-density lipoprotein cholesterol(F=102.197,P<0.01)levels,alleviate the phenomenon of massive lipid deposition in the liver,and increase the number of goblet cells in the small intestine.The results of intestinal fungi detection showed that there were 405 unique species in group Z,238 in group M,and 201 in group XS;the Chao1 index(F=22.070,P<0.01),ACE index(F=29.280,P<0.01),PD whole-tree index(F=211.000,P<0.01)and Pielou index(F=11.360,P<0.05)of rats in the spleen deficiency hyperlipidemia group decreased significantly,and the diversity of flora abundance decreased,while Xiangsha Liujunzi Decoction could reverse the diversity of intestinal fungal abundance in rats;at the phylum level,the ratio of Basidiomycota/Ascomycota in group M rats decreased significantly(F=4.824,P<0.05),which could be further reversed after administration of Xiangsha Liujunzi Decoction,but there was no significant difference;at the genus level,the fungal genera with higher percentage were Nectria,Cladosporium and Phaeosphaeria;the analysis of flora ecological function showed that the main nutritional mode of intestinal fungi in the three groups was saprophytic.Conclusion Xiangsha Liujunzi Decoction may improve dyslipidemia in rats with spleen deficiency and hyperlipidemia by altering the abundance and diversity of intestinal fungi.
作者
曹媛
冷雪
CAO Yuan;LENG Xue(Traditional Chinese Medicine Innovation Engineering Technology Center,Liaoning University of Traditional Chinese Medicine,Liaoning,Shenyang 110847,China;不详)
出处
《中国微生态学杂志》
北大核心
2025年第10期1117-1126,1136,共11页
Chinese Journal of Microecology
基金
国家自然科学基金青年科学基金(82104552)
辽宁省教育厅高校基本科研项目(LJ232410162027)
辽宁省科技计划联合计划(自然科学基金——面上项目)(2024-MSLH-293)。
关键词
脾虚高脂血症
肠道真菌
香砂六君子汤
ITS高通量测序
Spleen deficiency hyperlipidemia
Intestinal fungi
Xiangsha Liujunzi decoction
High-throughput sequencing
