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熊果酸对酒精性肝损伤大鼠肠道菌群的影响

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  • 国家科技期刊平台
  • NETL
  • NSTL
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  • 维普
目的:探究熊果酸(ursolic acid,UA)补充对酒精性肝损伤大鼠肠道菌群的影响,为酒精性肝损伤的防治拓展思路.方法:健康2 月龄雄性Wistar大鼠30 只随机分为3 组(每组10 只),分别为正常对照组(生理盐水)、酒精模型组(酒精)、UA干预组(酒精+UA),实验持续8 周.末次灌胃后12 h,麻醉后进行腹主动脉取血,同时留取肝脏组织及粪便.苏木精-伊红染色法观察各组大鼠肝脏组织病理学变化;赖氏法检测血清转氨酶活力;显色基质鲎试剂盒检测大鼠血浆内毒素水平;改良紫外酶促法检测血浆D-乳酸浓度;基因间重复共有序列(enterobacterial repetitive intergenic consensus,ERIC)-聚合酶链式反应(polymerase chain reaction,PCR)、PCR-变性梯度凝胶电泳(denaturing gradient gel electrophoresis,DGGE)技术获取粪便肠道菌群指纹图谱,并进行大鼠肠道菌群多样性分析;实时荧光定量PCR技术检测各组大鼠肠道内大肠杆菌、粪肠球菌、长双歧杆菌和嗜酸乳杆菌的含量.结果:苏木精-伊红染色病理学观察显示,酒精模型组大鼠肝小叶结构模糊,脂肪变性及炎性浸润增多;与酒精模型组相比,UA干预组肝组织损伤程度明显得到改善,且血清转氨酶活力明显降低(P＜0.05).此外,UA的补充显著抑制了酒精引起的血浆D-乳酸浓度及内毒素水平升高(P＜0.05).ERIC-PCR和PCR-DGGE结果显示,UA干预改善了酒精所致肠道菌群结构的紊乱,使其朝着正常组大鼠肠道菌群结构方向趋近.实时荧光定量PCR结果显示,UA干预组大鼠肠道内长双歧杆菌和嗜酸乳杆菌含量明显高于酒精模型组(P＜0.05),大肠杆菌和粪肠球菌含量较酒精模型组显著减少(P＜0.05),且均接近于正常对照组.结论:适量地补充UA能够有效改善酒精引起的大鼠肝脏损伤,其作用机制可能与UA调节酒精引起的肠道菌群结构和数量的改变、改善肠道微生态有关.
Effect of Ursolic Acid on Gut Microbiota in Rats with Alcohol-Induced Liver Injury
Objective: To explore the effect of ursolic acid (UA) on the gut microbiota of rats with alcoholic liver injury for the purpose of providing new insights into controlling alcoholic liver injury. Methods: A total of 30 healthy male Wistar rats at the age of 2 months were randomly divided into 3 groups with 10 animals in each group including normal control group (saline), alcohol model group and UA-treated group (alcohol + UA) throughout the 8-week experiment. The rats were weighed and anesthetized with 10% chloral hydrate at twelve hours after the last treatment. Then blood samples were collected by abdominal aorta puncture to determine biochemical parameters. Meanwhile, liver tissue and feces were also taken. Pathological changes of liver tissue were evaluated by hematoxylin-eosin (HE) staining. The activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were determined by the method of Reitman and Frankel. The endotoxin level in serum was tested by tachypleus amebocyte lysate kit. The content of D-lactic acid (D-LA) in plasma was determined by a modified enzymatic method at an ultraviolet wavelength. The genomic DNA of the intestinal flora was used as the template through enterobacterial repetitive intergenic consensus (ERIC)-polymerase chain reaction (PCR) amplification and PCR-denaturing gradient gel electrophoresis (DGGE). At the same time, the diversity of gut microbiota was analyzed in rats. Quantitative real-time PCR was used to quantify the changes of gut microbiota following ethanol and UA treatments. Results: The pathological results obtained by HE staining showed that the hepatic lobular structure in the alcohol model group was vague, and fatty degeneration and inflammatory infiltration were increased. Compared with the alcohol model group, liver tissue damage was significantly improved and the activity of serum transaminase was significantly reduced in the UA-treated group (P ＜ 0.05). In addition, D-LA and endotoxin levels in serum induced by alcohol were significantly suppressed after supplementing UA (P ＜ 0.05). The results of ERIC-PCR and PCR-DGGE indicated that the alcohol-induced disorder of the gut microbiota structure was brought back to almost normal after the supplementation of UA. The results of quantitative real-time PCR indicated that the numbers of Enterococcus faecalis and Escherichia coli in the UA-treated group were significantly decreased to almost normal levels when compared with the alcohol model group ( P＜0.05). Moreover, the numbers of Lactobacillus and Bifidobacterium were significantly higher in UA-treated group than in the alcohol model group (P ＜ 0.05). Conclusion: Supplementation of UA can effectively improve alcohol liver injury in rats, likely by modulating gut microbiota distribution and improving intestinal microecology.

ursolic acidalcohol-induced liver injurygut microbiotaD-lactic acidendotoxin

马浩然、贾海莲、张文龙、王 晶、张男男、李可欣、邰大鹏、戈 娜

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包头医学院 营养与食品健康研究所,内蒙古 包头 014040

包头医学院第一附属医院骨外科,内蒙古 包头 014010

包头医学院第二附属医院消化科,内蒙古 包头 014030

熊果酸 酒精性肝损伤 肠道菌群 D-乳酸 内毒素

国家自然科学基金地区科学基金内蒙古自治区教育厅高等学校"青年科技英才支持计划"项目青岛市博士后人员应用研究项目内蒙古自治区科技计划项目包头医学院博士科研启动基金内蒙古自治区研究生创新课题自治区级大学生创新创业训练计划项目

81760586NMYT-15-B112015140201602069BSJJ201630S201610127Y02201610133010

2018

10.7506/spkx1002-6630-201811023

食品科学
北京食品科学研究院

食品科学

CSTPCDCSCD北大核心EI
影响因子：1.327
ISSN：1002-6630
年,卷(期)：2018.39(11)
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