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 plantarum ZDY04 can inhibit TMAO induced thermogenesis by promoting reverse transport and decomposition of cholesterol in ApoE^(-/-) mice.
作者
汤景辉
秦嫚嫚
唐乐
魏华
裘梁
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)
出处
《中国微生态学杂志》
CAS
CSCD
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)。
