中国畜牧兽医 ›› 2022, Vol. 49 ›› Issue (1): 140-149.doi: 10.16431/j.cnki.1671-7236.2022.01.015

• 营养与饲料 • 上一篇    下一篇

DHA对高脂饲粮诱导肝脏脂肪积累的预防作用

安晓羽1,4, 郝伟2, 孙利婷1, 杨丹妮3, 常悦3, 赵婷3, 李康3, 李喻瞳3, 张智精3, 杨慧娣1   

  1. 1. 内蒙古医科大学基础医学院, 呼和浩特 010110;
    2. 内蒙古医科大学第三临床学院, 呼和浩特 010110;
    3. 内蒙古医科大学第一临床学院, 呼和浩特 010110;
    4. 湖北中医药高等专科学校, 荆州 434020
  • 收稿日期:2021-05-24 出版日期:2022-01-05 发布日期:2021-12-29
  • 通讯作者: 杨慧娣 E-mail:yanghuidi1980@163.com
  • 作者简介:安晓羽,E-mail:1025044036@qq.com;郝伟,E-mail:1347153283@qq.com。
  • 基金资助:
    国家自然基金地区基金项目(32160247);内蒙古自治区自然基金面上项目(2020MS03059);内蒙古自治区大学生创新创业一般项目(201910132007、202010132009);内蒙古医科大学实验室开放基金项目(2020ZN21);内蒙古医科大学大学生科技创新英才项目(YCPY20200019、YCPY20200043)

Preventive Effect of Docosahexaenoic Acid on Liver Lipid Accumulation Induced by High Fat Diet in Mice

AN Xiaoyu1,4, HAO Wei2, SUN Liting1, YANG Danni3, CHANG Yue3, ZHAO Ting3, LI Kang3, LI Yutong3, ZHANG Zhijing3, YANG Huidi1   

  1. 1. Basic Medical School, Inner Mongolia Medical University, Hohhot 010110, China;
    2. Third School of Clinical Medicine, Inner Mongolia Medical University, Hohhot 010110, China;
    3. First School of Clinical Medicine, Inner Mongolia Medical University, Hohhot 010110, China;
    4. Hubei College of Chinese Medicine, Jingzhou 434020, China
  • Received:2021-05-24 Online:2022-01-05 Published:2021-12-29

摘要: [目的] 探索二十二碳六烯酸(DHA)对高脂饲粮诱导肝脏脂肪积累的预防机制。[方法] 将32只雄性SPF级C57BL/6小鼠均分为4组,对照组(Con)饲喂普通饲粮,模型组(Model)饲喂高脂饲粮,DHA组分别在高脂饲粮中添加0.2 g DHA(DHAL)和1.0 g DHA(DHAH),饲喂周期为20周。饲喂期间每天称量体重及食物重量,计算摄食量;饲喂结束,采集肝脏和血液,ELISA法检测肝脏脂联素和血清甘油三酯(TG)的含量,实时荧光定量PCR检测新生脂肪合成关键酶(SREBP-1c、FAS)、脂肪酸氧化关键基因(PPARα、PPARγ、CPT-1A和ACOX)、线粒体基因(PGC-1α)、褐色脂肪化基因(Prdm16、UCP1)的表达,Western blotting法检测肝脏磷酸化ACC、AMPK和AKT蛋白的表达。[结果] 与Con组相比,Model组终体重、体脂重量和TG含量均显著增加(P<0.05),肝脏脂联素浓度显著降低(P<0.05)。与Model组相比,DHAL和DHAH组终体重、体脂重量和TG含量均显著降低(P<0.05),肝脏脂联素水平显著增加(P<0.05)。实时荧光定量PCR结果表明,与Con组相比,Model组SREBP-1c和FAS mRNA表达量均显著增加(P<0.05),PPARα、CPT-1A、ACOX、PGC-1α和UCP1 mRNA表达均显著降低(P<0.05);与Model组相比,DHAL和DHAH组SREBP-1c和FAS mRNA表达量均显著降低(P<0.05),且DHAH组表达量均显著低于DHAL组(P<0.05),DHAL和DHAH组PPARα、CPT-1A、ACOX、PGC-1α、Prdm16和UCP1 mRNA表达量均显著增加(P<0.05),DHAH组CPT-1A和ACOX mRNA表达量显著低于DHAL组(P<0.05),PPARγ的mRNA表达量在4组中没有显著差异(P>0.05)。Western blotting结果表明,Model组p-ACC显著高于Con和DHAH组,但显著低于DHAL组;Model组p-AMPK/AMPK和pAKT/AKT比值均显著低于Con组(P<0.05),DHAH组p-AMPK/AMPK和pAKT/AKT比值均显著高于Model和DHAL组(P<0.05)。[结论] DHA可降低高脂饲粮导致的C57BL/6小鼠终体重、体脂和TG含量的升高,增加肝脏脂联素的水平,促进脂肪酸氧化和白色脂肪细胞褐色化,从而预防肝脏的脂肪积累。

关键词: 高脂饲粮; 小鼠; 二十二碳六烯酸(DHA); 肝脏; 脂肪积累

Abstract: [Objective] This study was to explore the preventive mechanism of docosahexaenoic acid (DHA) on liver fat accumulation induced by high fat diet (HFD). [Method] 32 male mice were divided into 4 groups. Control group (Con) mice were fed with control diet, model group (Model) mice were feed with high fat diet, DHA groups were supplemented with 0.2 g DHA (DHAL) and 1.0 g DHA (DHAH) in high fat diet, respectively. The feeding cycle was 20 weeks. The body weight and diet weight were weighed during feeding, and the food intake was calculated. At the end of feeding, liver and blood were collected, and the contents of liver adiponectin and serum triglyceride (TG) were detected by ELISA. The expression of key enzymes of newborn lipogenesis (SREBP-1c, FAS), key genes of fatty acid oxidation (PPARα, PPARγ, CPT-1A and ACOX), mitochondrial gene (PGC-1α) and brown fatty genes (PRDM16, UCP1) were detected by Real-time quantitative PCR. The expression of the hepatic phosphorylated ACC, AMPK and AKT proteins were determined by the Western blotting. [Result] Compared with Con group, the final body weight, body fat weight and TG content were significantly increased (P<0.05), and hepatic adiponectin level was significantly decreased in Model group (P<0.05). When compared with Model group, the final body weight, body fat weight and TG content of DHAL and DHAH groups were significantly decreased (P<0.05), and liver adiponectin level was significantly increased(P<0.05). The results of Real-time quantitative PCR showed that compared with Con group, SREBP-1c and FAS mRNA level of Model group were significantly increased (P<0.05), PPARα, CPT-1A, ACOX, PGC-1α and UCP1 mRNA level of Model group were significantly decreased (P<0.05). While compared with Model group, SREBP-1c and FAS mRNA levels of both DHAH and DHAL groups were significantly decreased (P<0.05), and the level of DHAH group was lower than that of DHAL group (P<0.05), PPARα, CPT-1A, ACOX, PGC-1α, Prdm16 and UCP1 mRNA expression level of DHAL and DHAH groups were significantly increased (P<0.05), CPT-1A and ACOX mRNA level of DHAH group was higher than that of DAHL group (P<0.05), and there was no significant difference in PPARγ mRNA expression level among the four groups(P>0.05). The results of Western blotting showed that p-ACC in Model group was significantly higher than that in Con and DHAH groups, but significantly lower than that in DHAL group, p-AMPK/AMPK and p-AKT/AKT of Model group were smaller than those of Con group (P<0.05), p-AMPK/AMPK and p-AKT/AKT of DHAH group were bigger than those of Model and DHAL groups (P<0.05). [Conclusion] DHA could reduce the increase of final body weight, body fat and TG content in C57BL/6 mice caused by high fat diet, promote fatty acid oxidation and browning of white adipocytes, so as to prevent fat accumulation in liver.

Key words: high fat food; mice; docosahexaenoic acid (DHA); liver; lipid accumulation

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