植物乳杆菌ZDY04促进胆固醇逆向转运减轻TMAO诱导的ApoE^(-/-)小鼠动脉粥样硬化  

Lactobacillus plantarum ZDY04reduces trimethylamine N-oxide-induced atherosclerosis by promoting reverse cholesterol transport in ApoE^(-/-)mice

作　　者：汤景辉 秦嫚嫚 唐乐 魏华[3] 裘梁[1,2] 余军 TANG Jing-hui;QIN Man-man;TANG Le;WEI Hua;QIU Liang;YU Jun(Translational Medicine Center,Jiangxi University of Traditional Chinese Medicine,Nanchang,Jiangxi 330006,China;Jiangxi Key Laboratory of Traditional Chinese Medicine for Prevention and Treatment of Vascular Remodeling Related Diseases,Jiangxi University of Traditional Chinese Medicine,Nanchang,Jiangxi 330006,China;不详)

机构地区：[1]江西中医药大学医学转化中心,江西南昌330006 [2]江西中医药大学江西省中药防治血管重塑相关疾病重点实验室 [3]南昌大学食品与科学技术国家重点实验室 [4]Department of Physiology and Center for Metabolic Disease Research,Lewis Katz School of Medicine,Temple University,Philadelphia,PA19140,USA

出　　处：《中国微生态学杂志》2022年第3期249-256,共8页Chinese Journal of Microecology

基　　金：国家自然科学基金(81860090);江西省教育厅一般项目(GJJ190677);江西中医药大学博士启动基金(2019WBZR009);江西省自然科学基金(20202BABL206007);江西省重点实验室建设项目(20202BCD42014);南昌市科技局项目(2018-NCZDSY-005)。

摘　　要：目的探讨植物乳杆菌ZDY04能否改善氧化三甲胺(TMAO)诱导的小鼠动脉粥样硬化及其机制。方法6~8周龄雌性载脂蛋白E基因敲除(apolipoprotein E-deficient mice,ApoE^(-/-))小鼠被随机分成3组:正常饮食组(Chow组)、胆碱+溶剂组(Choline+PBS组)和Choline+ZDY04组,每组5只,通过自由饮用1.3%高胆碱饮食构建小鼠动脉粥样硬化模型,灌胃含有15%甘油的无菌PBS或菌株,持续16周,检测血脂水平,uHPLC-MS/MS测定血浆TMAO和粪便三甲胺(TMA)的含量,采用油红“O”染色检测全长主动脉粥样硬化病变程度,采用苏木素-伊红、油红“O”以及Masson染色分别检测主动脉根部斑块面积、脂质占比以及胶原纤维含量,采用免疫荧光检测主动脉根部斑块中巨噬细胞以及平滑肌细胞数量,采用RT-qPCR和Western blotting检测小鼠肝脏胆固醇逆向转运相关基因及Fmo3基因的表达水平。结果与Choline+PBS组相比,Choline+ZDY04组小鼠主动脉脂质沉积区域显著减少,主动脉根部动脉粥样硬化斑块面积、脂滴及巨噬细胞阳性区域显著减少,胶原纤维和平滑肌细胞在斑块中占比没有显著变化。粪便TMA及血清TMAO含量显著降低。血清总胆固醇、低密度脂蛋白胆固醇含量具有下降趋势,但差异无统计学意义,高密度脂蛋白胆固醇含量显著上升。胆固醇逆向转运相关基因Sr-b1、Abcg5和Cyp7a1等显著升高,Fmo3基因表达水平没有变化。结论植物乳杆菌ZDY04通过促进胆固醇的逆向转运和分解,抑制ApoE^(-/-)小鼠体内的动脉粥样硬化。Objective To observe the effect of Lactobacillus plantarum ZDY04 in trimethylamine N-oxide(TMAO)-induced atherosclerosis in mice and its underlying mechanisms. Methods Fifteen female(six to eight-weeks old) apolipoprotein E-deficient(ApoE^(-/-)) mice were randomly assigned into three groups as follows: Chow group, Choline+PBS group and Choline+ZDY04 group, and fed with 1.3% high-choline diet to establish atherosclerosis models, then treated with sterile PBS or ZDY04 containing 15% glycerol for 16 weeks. After treatment, serum lipids were measured using cholesterol kit. The serum TMAO and fecal TMA content were determined by using liquid chromatography-tandem mass spectrometry LC-MS. Atherosclerotic lesion formation in whole aorta was detected with Oil red O staining. Hematoxylin-eosin, Oil red O and Masson staining were used to detect the atherosclerotic area, lipid deposition and collagen content in the aortic roots. Immunofluorescent staining of macrophage specific antigen CD68 and smooth muscle a-actin(SMA) were used to examine the lesion composition in the aortic root. The expression of genes related to reverse cholesterol transport and Fmo3 in liver was detected with RT-qPCR and Western blotting. Results Compared with Choline+PBS group, the atherosclerotic lesions in the whole aortas significantly reduced in Choline+ZDY04 group. ZDY04 treatment significantly decreased atherosclerotic lesion size, lipid deposition and macrophage content, but not the contents of lesion smooth muscle cells or collagen, when compared to Choline+PBS fed mice. The content of cecum TMA and serum TMAO decreased significantly in Choline+ZDY04 group;serum high density lipoprotein-cholesterol significantly increased, but total cholesterol and low density lipoprotein-cholesterol only marginally decreased. ZDY04 significantly increased the mRNA expression of reverse cholesterol transport genes Sr-b1, Abcg5, Cyp7a1 but not Fmo3. Liver CYP7 A1 protein level was also increased by ZDY04 compared to controls. Conclusion Lactobacillus plantaru

关 键 词：动脉粥样硬化 植物乳杆菌ZDY04 氧化三甲胺 胆固醇逆向转运 

分 类 号：Q93[生物学—微生物学]