目的 探讨苹果多酚提取物 (apple polyphenol extracts , APE) 对葡聚糖硫酸钠 (dextran sodium sulfate,DSS) 诱导的急性溃疡性结肠炎 (ulcerative colitis, UC) 相关肝脏脂代谢的影响及可能的机制。方法 30只6~8 w龄雄性C57BL/6小鼠随机分为对照组 (CON)、模型组 (DSS)、APE干预组 (AD),给与标准饲料喂养,其中,CON组正常饮水,DSS组前5 w正常饮水,第6 w 3% DSS饮水;AD组给与500 mg/ (kg·bw·d) APE连续灌胃6 w,第6 w同时给与3% DSS饮水。干预结束后,小鼠禁食过夜,眼眶后静脉丛取血后处死,取结肠、肝脏,并观察结肠远端、左侧肝小叶组织病理学变化。采用实时定量聚合酶链式反应 (quantitative real time polymerase chain reaction, RT-qPCR) 检测肝脏脂代谢相关基因mRNA的表达水平。结果 APE干预显著降低UC小鼠的疾病活动指数 (disease activity index, DAI)评分,改善结肠病理学损伤,降低病理学评分;同时改善肝脏脂质沉积和炎细胞浸润,降低非酒精性脂肪肝病(non-alcoholic fatty liver disease, NAFLD)活动度积分 (NAFLD activity score, NAS),并显著降低血清甘油三酯 (triglyceride, TG)、谷草转氨酶 (aspartate aminotransferase,AST)、谷丙转氨酶 (alanine aminotransferase,ALT)以及肿瘤坏死因子-α (tumor necrosis factor-α, TNF-α)、白细胞介素6 (interleukin-6, IL-6) 和白细胞介素1β(interleukin-1β, IL-1β) 水平。进一步的mRNA检测结果显示,与CON组相比,DSS组小鼠肝脏脂肪酸氧化相关基因过氧化物酶体增殖物激活受体α (peroxisome proliferators -activated receptor α, Pparα)、脂肪酸摄取基因白细胞分化抗原36 (cluster of differentiation 36, Cd36)、脂肪分解基因脂肪甘油三酯脂肪酶 (adipose triglyceride lipase, Atgl)、胆固醇逆转运基因ATP 结合盒亚家族 G 成员 8 (ATP binding cassette subfamily G member 8, Abcg8)、胆汁酸合成基因胆固醇7α-羟化酶（cholesterol 7-α hydroxylase, Cyp7a1) 以及肝脏脂代谢关键调节因子沉默信息调节因子 (silent information regulator 1, Sirt1) mRNA表达水平显著降低 (P<0.05)。肝脏中脂肪生成调控基因乙酰辅酶A羧化酶 (acetyl CoA carboxylase, Acc)、胆汁酸受体胆汁酸G蛋白偶联受体5 (G protein-coupled bile acid receptor 5, Tgr5)、促炎细胞因子Tnf-α以及Il-6的表达相显著上升(P<0.05)。与DSS组相比,APE干预显著上调Sirt1、胆固醇逆转运基因B类1型清道夫受体 (scavenger receptor class B type 1, Sr-b1)、肝脏X受体α (liver X receptor α, Lxrα) mRNA表达水平,显著降低了Acc、Tgr5、Tnf-α mRNA表达(P<0.05)。结论 APE通过调节肝脏脂代谢相关基因的表达改善急性UC相关肝脏脂代谢失调。
Abstract
Objective To investigate the effects of apple polyphenol extract (APE) on the changes of hepatic lipid metabolism induced by dextran sodium sulfate (DSS) in mice with acute ulcerative colitis (UC) and the underlying mechanisms. Methods Thirty male C57BL/6 mice aged 6-8 weeks were randomly divided into three groups, namely control (CON), DSS model (DSS) and DSS model treated with APE (AD). All mice were fed the chow diet. Mice in CON group were given distilled water for 6 weeks. For mice in the DSS group, distilled water was provided for the first 5 weeks and 3% DSS (m/v) solution in drinking water was given for the last week. For mice in the AD group, distilled water was provided provided for the first 5 weeks and 3% DSS (m/v) solution in drinking water was given for the last week, simultaneously with 500 mg/(kg·bw) APE gavage for 6 weeks. After the intervention, blood samples from the angular vein were obtained, while colon and liver were collected for subsequent histological examination. Quantitative real time polymerase chain reaction (RT-qPCR) was employed to detect the mRNA expression of genes related to hepatic lipid metabolism. Results APE intervention significantly reduced DAI scores in UC mice, ameliorated colonic pathological damages, diminished hepatic lipid deposition and inflammatory cell infiltration, reduced NAS, and along with significantly reduced serum levels of TG, AST, ALT, as well as levels of TNF-α, IL-6 and IL-1β. Further mRNA analysis showed that compared with the CON group, the mRNA expression levels of the fatty acid oxidation-related genes Pparα, fatty acid uptake gene Cd36, lipolysis gene Atgl, cholesterol reverse transporter genes Abcg8, bile acid synthesis genes Cyp7a1 and the key regulator of hepatic lipid metabolism Sirt1 were significantly decreased (P<0.05). The mRNA levels of Acc, Tgr5, proinflammatory cytokine Tnf-α and Il-6 were significantly increased (P<0.05). Compared with the DSS group, APE intervention significantly increased the mRNA expression levels of Sirt1, Sr-b1 and Lxrα (P<0.05),reduced the mRNA expression levels of Acc, Tgr5 and Tnf-α (P<0.05). Conclusion APE improves acute UC-associated hepatic lipid metabolism abnormalities by regulating the mRNA levels of hepatic lipid metabolism-related genes.
关键词
苹果多酚 /
DSS /
急性溃疡性结肠炎 /
肝脏 /
脂代谢
Key words
apple polyphenols extract /
dextran sodium sulfate /
acute ulcerative colitis /
liver /
lipid metabolism
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