摘要
目的:观察桂枝汤对高脂饮食导致的载脂蛋白E基因敲除(ApoE^(-/-))小鼠外周血单核细胞、肠道菌群和主动脉血管动脉粥样硬化(AS)斑块形成的影响。方法:40只12周龄雌性ApoE^(-/-)小鼠采用配对比较法随机分为普食组(ApoE^(-/-)+CD),高脂组(ApoE^(-/-)+WD),桂枝汤组(ApoE^(-/-)+WD+GZT,7.83 g·kg^(-1))和阿托伐他汀组(ApoE^(-/-)+WD+Atr,3.33 mg·kg^(-1)),10只匹配的C57BL/6小鼠为野生普食组(C57BL/6+CD)。除普食组外,其余给予高脂饮食诱导AS模型。治疗组于高脂饮食同时分别给予口服灌胃桂枝汤和阿托伐他汀,正常组和模型组采用同时间、同体积双蒸水灌胃对照。干预4周,每组取5只小鼠,取眼球血,采用全自动生化分析仪检测血浆血脂水平、流式细胞仪检测外周血单核细胞及其亚型比例、表面受体Toll样受体4(TLR4)和CD36表达水平;取回肠内容物采用16S rRNA相对应的DNA序列(16S rDNA)测序检测小鼠肠道菌群。每组余下的5只小鼠干预12周后,取胸腹主动脉采用油红O染色检测主动脉血管斑块形成情况。结果:干预4周,与C57BL/6+CD组比较,ApoE^(-/-)+CD组血浆总胆固醇(TC)和低密度脂蛋白(LDL)水平升高(P<0.01),ApoE^(-/-)+WD组血浆TC和LDL水平进一步升高(P<0.01);ApoE^(-/-)+WD组外周血单核细胞及其炎症亚型Ly6C比例升高,TLR4表达上升(P<0.05);ApoE^(-/-)+WD组回肠菌群的厚壁菌门增加,疣微球菌门减少。与ApoE^(-/-)+WD组比较,ApoE^(-/-)+WD+GZT组血脂水平没有明显改变,但炎症亚型单核细胞Ly6C++比例降低,单核细胞表面受体TLR4和CD36表达降低(P<0.05);回肠菌群的厚壁菌门比例减少,拟杆菌门和疣微球菌门增加。干预12周,ApoE^(-/-)+WD组主动脉血管AS斑块形成,ApoE^(-/-)+WD+GZT组AS斑块病变程度减轻。结论:桂枝汤可以改善高脂饮食饲喂ApoE^(-/-)小鼠的单核细胞免疫异常以及肠道菌群失衡,抑制AS斑块形成。
Objective: To observe the effect of Guizhitang(GZT)on peripheral blood monocytes,intestinal flora and AS plaque formation of ApoE^(-/-)mice induced by Western diet(WD). Method: In this study,40 12-week-old homozygous female ApoE^(-/-)mice were randomly divided into chow diet(CD)group(ApoE^(-/-)+CD),WD group(ApoE^(-/-)+WD),GZT group(ApoE^(-/-)+WD+GZT,7.83 g·kg^(-1))and atorvastatin(Atr)group(ApoE^(-/-)+WD+Atr,3.33 mg·kg^(-1)). And 10 matched C57 BL/6 mice were set as wild CD control group(C57 BL/6+CD). Except the CD group,the rest groups were given WD to induce models. Treatment groups were given Guizhitang or atorvastatin orally in addition to WD,while ApoE^(-/-)+CD and ApoE^(-/-)+WD model groups were treated with the same volume of double steam water at the same time. After 4 weeks of intervention,5 mice in each group were selected to collect the eyeball blood samples. The levels of plasma lipids were detected by automatic biochemical analyzer,and the proportion of peripheral blood mononuclear cells and its subtypes,and the expression levels of surface receptors toll like receptor 4(TLR4)and CD36 were detected by flow cytometry,the intestinal flora of mice was detected by 16 S rDNA sequencing. The remaining 5 mice in each group were intervened for 12 weeks,and the aorta was taken to detect the formation of aortic plaque by oil red O staining. Result:After intervention for 4 week,compared with C57 BL/6+CD group,the levels of plasma total cholesterol(TC)and low-density lipoprotein(LDL)levels in ApoE^(-/-)+CD and ApoE^(-/-)+WD groups were increased(P<0.01). ApoE^(-/-)+WD group showed increase in the proportion of monocytes,their inflammatory subtypes Ly6 C,and TLR4 expression on monocyte surface in blood(P<0.05). ApoE^(-/-)+WD group induced the imbalance of intestinal flora, with increase of Firmicutes and decrease of Verrucomicrobia in ileum of ApoE^(-/-)mice. Compared with ApoE^(-/-)+WD group,there was no significant change in blood lipid level and monocyte proportion in ApoE^(-/-)+WD+GZT group,but with decrease in the proportion of Ly6 C,increase in the proportion of anti-inflammatory subtype Ly6 C-,and decrease in the expression of TLR4 and CD36 on monocyte surface(P<0.05). ApoE^(-/-)+WD+GZT group showed decrease of Firmicutes and increase of Bacteroidetes and Verrucomicrobia in ileum of ApoE^(-/-)mice. After 12 weeks of intervention,ApoE^(-/-)+WD group showed increase in the number and area of aortic plaques in ApoE^(-/-)mice. ApoE^(-/-)+WD+GZT group showed decrease of the area of aortic AS plaques. Conclusion:GZT can reduce the immune damage and imbalance of intestinal flora caused by WD,then inhibit the formation of AS plaque.
作者
袁晓雯
姜楠
柏冬
陈冰
李玉梅
曾辉
马雅銮
YUAN Xiao-wen;JIANG Nan;BAI Dong;CHEN Bing;LI Yu-mei;ZENG Hui;MA Ya-luan(Institute of Basic Theory,China Academy of Chinese Medical Sciences,Beijing 100700,China;Institute of Infectious Disease,Beijing Ditan Hospital,Capital Medical University,Beijing 100015,China;Beijing Key Laboratory of Emerging Infectious Diseases,Beijing 100015,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2021年第4期24-29,共6页
Chinese Journal of Experimental Traditional Medical Formulae
基金
中国中医科学院自主选题项目(YZ-1702)。
