摘要
目的探讨动脉粥样硬化(动脉硬化)合并血清诱导型炎性关节炎小鼠肠道菌群及免疫系统的变化。方法实验分为ApoE基因缺陷(ApoE^-/-)组和C57BL/6对照组,通过给8周龄ApoE^-/-小鼠和C57BL/6小鼠分别饲喂高脂食物,ELISA法检测血脂蛋白水平,油红O染色来评价动脉硬化斑块面积,从而评估小鼠患动脉硬化的严重程度。从17周龄起腹腔注射K/BxN血清至26周龄,游标卡尺测量小鼠踝关节宽度,通过踝关节宽度和血清诱导型炎性关节炎评分来评价小鼠患血清诱导型炎性关节炎的程度。取小鼠粪便并提取DNA进行16S V4区PCR扩增,Illumina HiSeq PE250上机测序。分离小鼠脾脏细胞和骨髓细胞并用流式抗体染色,经流式细胞仪分析免疫细胞分布的改变。结果与C57BL/6对照组相比,动脉硬化合并血清诱导型炎性关节炎小鼠血脂水平显著升高,动脉硬化斑块面积显著增加;踝关节宽度和血清诱导型炎性关节炎评分降低;肠道菌群多样性降低,疣微菌纲显著降低;脾脏和骨髓CD3^+T细胞和CD19^+B细胞显著降低,CD11b^+巨噬细胞显著升高。结论动脉硬化合并血清诱导型炎性关节炎引起小鼠免疫细胞分布改变,肠道菌群多样性降低,细菌菌群分布改变,健康肠道的潜在生物标志物艾克曼菌丰度降低。推测肠道微生物的改变可能参与动脉硬化合并血清诱导型炎性关节炎的发病机制。
Objective To investigate the changes in intestinal microbiota and immune cells in mice with atherosclerosis and arthritis.Methods Two groups were set up with 8-week-old ApoE knockout(ApoE^-/-)and C57BL/6 mice.All mice were fed with high fat diet.The severity of atherosclerosis was evaluated through analyzing the level of serum lipid by ELISA and arteriosclerotic plaques by oil red O staining.K/BxN serum was intraperitoneally injected into ApoE^-/-and C57BL/6 mice at 17 to 26 weeks of age until 26 weeks to induce arthritis.The development of arthritis in mice was evaluated with vernier caliper-measured ankle width and clinical score.Total DNA was extracted from fecal samples of mice to amplify the 16S V4 region by PCR,and then sequencing analysis was performed on Illumina HiSeq PE250.Splenocytes and bone marrow cells were isolated and stained by antibodies to analyze the changes in immune cell distribution by flow cytometry.Results The levels of low density lipoprotein cholesterol(LDL-C)and total cholesterol(TCHO)in serum and the atherosclerotic plaques in aorta were significantly increased in ApoE^-/-mice than in C57BL/6 mice,suggesting that the symptoms of atherosclerosis in ApoE^-/-mice were more severe than those in C57BL/6 mice.Ankle width measurements and clinical scores of arthritis showed that the ApoE^-/-mice had less severe ankle swelling than the C57BL/6 mice.Moreover,the ApoE^-/-mice had decreased intestinal microbiota diversity,especially in the number of Verrucomicrobiae.Flow cytometry analysis showed that CD3^+T and CD19^+B cells were down-regulated,while CD11b^+macrophages were up-regulated in splenocytes and bone marrow cells of the ApoE^-/-mice.Conclusions Immune cell distribution was changed in the ApoE^-/-mice with atherosclerosis and arthritis.Compared with the C57BL/6 mice,the ApoE^-/-mice had decreased intestinal microbiota diversity and altered bacterial composition.Moreover,the abundance of Akkermansia,a potential biomarker of healthy intestinal environment,in the ApoE^-/-mice was also dec
作者
石娜
张抒扬
牛海涛
Shi Na;Zhang Shuyang;Niu Haitao(NHC Key Laboratory of Human Disease Comparative Medicine (the Institute of Laboratory Animal Sciences,CAMS&PUMC),Beijing Key Laboratory for Animal Models of Emerging and Reemerging Infectious,Beijing Engineering Research Center for Experimental Animal Models of Human Critical Diseases,Key Laboratory of Human Diseases Animal Model,State Administration of Traditional Chinese Medicine,Beijing 100021,China;Department of Cardiology,Peking Union Medical College Hospital,Beijing 100010,China)
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2019年第4期283-291,共9页
Chinese Journal of Microbiology and Immunology
基金
国家重点研发计划资金资助(2017YFC1103603)
中国医学科学院医学与健康科技创新工程资金资助(2016-12M-1-006).
