壮药玉郎伞对动脉粥样硬化小鼠肠道菌群及其代谢产物的影响

Effects of Zhuang Medicine Yulangsan on Intestinal Flora and Its Metabolites in Mice with Atherosclerosis

关键词:  玉郎伞  动脉粥样硬化  肠道菌群  代谢产物

Key words:  Yulangsan  atherosclerosis  intestinal flora  metabolites

摘要:

为观察壮药玉郎伞对动脉粥样硬化(Atherosclerosis,AS)小鼠肠道菌群及其代谢产物的影响，分析其抗AS的潜在作用机制，将40只SPF级ApoE^-/-(Apolipoprotein E knockout)雄性小鼠随机分为模型(Model)组、阿托伐他汀(Ato)组以及玉郎伞高、中、低剂量（YLS-H、YLS-M、YLS-L）组，每组各8只，另选取８只健康雄性C57BL/6J小鼠作为空白对照(Normal)组。除Normal组外，其余5组均采用高脂饲料喂养12周构建AS模型，造模成功后按组别灌胃给药，Normal组、Model组灌胃等量无菌0.9%氯化钠溶液，Ato组给药剂量为5.0 mg/(kg·d)，YLS-H组、YLS-M组、YLS-L组给药剂量分别为10.0、5.0、2.5 mg/(kg·d)，连续给药8周后取材，检测其血清甘油三酯(Triglyceride,TG)、总胆固醇(Total Cholesterol,TC)、低密度脂蛋白胆固醇(Low-Density Lipoprotein Cholesterol,LDL-C)、高密度脂蛋白胆固醇(High-Density Lipoprotein Cholesterol,HDL-C)水平，并分析其粪便肠道菌群结构和差异代谢物。结果显示，与Model组相比，YLS组可剂量依赖性降低主动脉粥样硬化斑块面积占比，下调TG、TC、LDL-C水平，上调HDL-C （P<0.01）水平；YLS-H组调节紊乱的菌群Alpha多样性，使门、科、属水平菌群结构趋向正常水平，上调厚壁菌门（Firmicutes）等有益菌丰度(P<0.05)；与Normal组、YLS组相比，Model组共筛选出21个主要富集于类固醇激素生物合成、甘油磷脂代谢等通路的潜在差异代谢物；玉郎伞干预可改善上述差异代谢物的异常水平。综上所述，壮药玉郎伞可抑制AS小鼠主动脉粥样硬化斑块形成，减轻组织病理损伤，改善血脂紊乱，其作用机制可能与改善肠道差异菌群及其相关代谢通路有关。

Abstract:

This study observed the effects of Zhuang medicine Yulangsan (YLS) on intestinal flora and its metabolites in mice with atherosclerosis (AS),aiming to explore the potential mechanism underlying the anti-atherosclerotic activity of YLS.Forty male SPF-grade apolipoprotein E knockout (ApoE^-/-) mice were randomly assigned into 5 groups (8 mice per group):a Model group,an atorvastatin (Ato) group,a high-dose YLS (YLS-H) group,a medium-dose YLS (YLS-M) group,and a low-dose YLS (YLS-L) group.Another 8 healthy male C57BL/6J mice were set as the Normal group.Mice in the other 5 groups except the Normal group were fed a high-fat diet for 12 weeks to establish the model of AS.After successful modeling,gavage was performed according to grouping.Mice in the Normal and Model groups received an equal volume of sterile 0.9% sodium chloride solution via gavage.The Ato group was treated with atorvastatin at 5.0 mg/(kg·d).The YLS-H,YLS-M,and YLS-L groups were administered YLS at doses of 10.0,5.0,and 2.5 mg/(kg·d),respectively.After continuous administration for 8 weeks,samples were collected.The serum levels of Triglyceride (TG),Total Cholesterol (TC),Low-Density Lipoprotein Cholesterol (LDL-C),and High-Density Lipoprotein Cholesterol (HDL-C) were measured.In addition,the intestinal flora structure and differential metabolites in feces were analyzed.The results showed that compared with the Model group,YLS reduced the proportion of aortic plaque area in a dose-dependent manner,down-regulated the levels of TG,TC,and LDL-C,and up-regulated the level of HDL-C (P<0.01).YLS-H restored the disordered alpha diversity of intestinal flora,shifted the phylum- family- and genus-level microbial community structure toward the normal state,and increased the relative abundance of beneficial bacteria such as Firmicutes (P<0.05).Compared with the Normal and YLS groups,a total of 21 potential differential metabolites were screened in the Model group,which were mainly enriched in pathways including steroid hormone biosynthesis and glycerophospholipid metabolism.Intervention with YLS restored the levels of the above differential metabolites.In conclusion,the Zhuang medicine YLS inhibits the formation of atherosclerotic plaques,alleviates histopathological injury,and ameliorates dyslipidemia in AS mice by regulating the differential taxa in intestinal flora and their associated metabolic pathways.