基于血清代谢组学研究黄芪甲苷抑制高脂喂养的Apc^(Min/+)小鼠结肠腺瘤性息肉形成  

Astragalaside IV Inhibits Colonic Adenomatous Polyps Formation in High-fat Diet-fed Apc^(Min/+)Mice by Regulating Serum Metabolomics

作　　者：贺秋兰[1] 温鲁平 李贵明 林中原 高凤娇 黄文起[1] HE Qiu-lan;WEN Lu-ping;LI Gui-ming;LIN Zhong-yuan;GAO Feng-jiao;HUANG Wen-qi(Department of Anesthesiology,The First Affiliated Hospital of Sun Yat-sen University,Guangzhou 510080,China;Anorectal Section,Affiliated Hospital of Xuzhou Medical University,Xuzhou,221000,China;Department of Colorec‐tal and Anal Surgery,The Sixth Affiliated Hospital of Sun Yat-sen University,Guangzhou,510655,China)

机构地区：[1]中山大学附属第一医院麻醉科,广东广州510080 [2]徐州医科大学附属医院肛肠科,徐州江苏221000 [3]中山大学附属第六医院结直肠肛门外科,广州广东510655

出　　处：《中山大学学报（医学科学版）》2022年第6期916-927,共12页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(82004368)。

摘　　要：【目的】探究黄芪甲苷(ASIV)对高脂喂养(HF)的腺瘤性息肉病基因突变(Apc^(Min/+))小鼠结肠腺瘤性息肉(CAP)形成的预防作用及对血清代谢物紊乱的调控机制。【方法】将Apc^(Min/+)小鼠分成对照组(Con)、高脂喂养组(HF)、黄芪甲苷治疗组(HF-ASIV),每组8只。HF组和HF-ASIV组喂养高脂饲料8周,HF-ASIV组同时每两天给予ASIV(50 mg/kg)灌胃。记录小鼠每日饮水量、进食量和体质量变化。8周末收集小鼠血清,取小肠末端及近端结肠记录CAP个数和病理观察(HE);免疫组化(IHC)检测各组小鼠CAP周围肠壁COX2水平;qRT-PCR检测肠壁IL-1β和TNFα表达水平;液相质谱技术对小鼠肠道进行非靶向代谢轮廓分析,筛选差异代谢物并进行功能富集。【结果】实验期间各组小鼠无死亡,HF-ASIV组与HF组相比肠息肉总数减少,以中腺瘤(直径1~3 mm)、大腺瘤(直径>3 mm)减少为主。HF-ASIV组小鼠肠壁的杯状细胞及潘氏细胞数量较HF组增加;肠壁COX2、IL-1β和TNFα表达水平下降。组间两两比较的代谢物OPLS-DA评分差异显著(P<0.001;P=0.02)。与HF组比较,HF-ASIV组血清中有13种差异代谢物发生显著回调,其中正负离子模式下功能富集到N-乙酰-α-D-氨基葡萄糖基-二磷酸的代谢物水平均显著提高(P<0.05),并与腺瘤数量呈负相关(P<0.01)。【结论】ASIV可减少高脂喂养Apc^(Min/+)小鼠结肠腺瘤性息肉的数量,与降低肠壁炎症、调节血清代谢产物N-乙酰-α-D-氨基葡萄糖基-二磷酸水平密切相关。【Objective】To investigate the preventive effects of Astragaloside IV(ASIV)on the formation of colonic ad-enomatous polyps(CAP)in high-fat-fed(HF)mice with adenomatous polyposis mutation(Apc^(Min/+))and its regulation mechanism on serum metabolite disorders.【Methods】A total of 24 mice were evenly assigned to Apc^(Min/+)mice group(Con,n=8),high-fat diet-fed Apc^(Min/+)mice group(HF,n=8),and ASIV treatment group(HF-ASIV,n=8).Apc^(Min/+)mice in HF group and HF-ASIV group were given high-fat diet for 8 consecutive weeks to construct CAP mouse model.Mean-while,Apc^(Min/+)mice in HF-ASIV group were given 50 mg/kg ASIV every other day for 8 weeks.Daily amount of water in-take,food intake and body weight of mice in each group were recorded.At the end of the 8th week,mice were sacrificed for serum and the distal and proximal colon.The number of colonic polyps in each group was recorded.Hematoxylin-eosin(H&E)staining was used to observe the morphological changes of the proximal colon.Immunohistochemistry(IHC)was applied to analyze the amount of COX2 positive cells in the proximal colon.The expression levels of IL-1βand TNFαin intestinal wall were detected by QRT PCR.Non-targeted metabolic profiling of mouse gut by liquid-phase mass spectrome-try was used to screen for differential metabolites and perform functional enrichment.【Results】No mice died in each group during experiment.ASIV effectively reduced the size and the number of the CAPs that accompanied with the downregulation of inflammatory signaling molecules(COX-2,IL-1βand TNFα).Alcian blue and lysozyme staining showed ASIV im-proved the gut epithelium by promoting goblet and Paneth cells population.Significant differences in metabolite OPLS-DA scores between groups were noted(P<0.001;P=0.02).Compared with HF group,13 differential metabolites in the se-rum of HF-ASIV group were significantly recalled,and the levels of the metabolites functionally enriched to N-acetyl-αD-glucosaminyl-diphosphate were significantly increased in the positive and negative

关 键 词：腺瘤性息肉 黄芪甲苷 结肠 代谢组学 N-乙酰-α-D-氨基葡萄糖基 

分 类 号：R735.35[医药卫生—肿瘤]