研究论文 RESEARCH PAPER

银杏叶提取物通过脂联素/脂联素受体1/腺苷酸激活蛋白激酶信号通路改善肥胖犬脂质代谢及脂肪沉积的研究

  • 陆江 ,
  • 朱道仙 ,
  • 卢劲晔 ,
  • 刘莉 ,
  • 刘静
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  • 1. 江苏农牧科技职业学院宠物科技学院, 泰州 225300;
    2. 江苏农牧科技职业学院动物医学院, 泰州 225300
陆江(1982—),男,江苏新沂人,副教授,硕士,主要从事小动物临床营养与代谢病研究。E-mail:vetlj@163.com

收稿日期: 2021-09-27

  网络出版日期: 2022-05-14

基金资助

江苏省高校教师青蓝工程学科带头人项目(2021);江苏农牧科技职业学院校级课题(NSF2021ZR08)

Ginkgo biloba Extract Improves Lipid Metabolism and Fat Deposition of Obese Dogs via Adiponectin/Adiponectin Receptor 1/Adenosine Monophosphate-Activated Protein Kinase Signaling Pathway

  • LU Jiang ,
  • ZHU Daoxian ,
  • LU Jinye ,
  • LIU Li ,
  • LIU Jing
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  • 1. Department of Pet Science and Technology, Jiangsu Agri-Animal Husbandry Vocational College, Taizhou 225300, China;
    2. Department of Animal Medicine, Jiangsu Agri-Animal Husbandry Vocational College, Taizhou 225300, China

Received date: 2021-09-27

  Online published: 2022-05-14

摘要

本试验旨在研究脂联素(APN)/脂联素受体1(AdipoR1)/腺苷酸激活蛋白激酶(AMPK)信号通路在银杏叶提取物改善高脂饮食诱导的肥胖犬脂质代谢及脂肪沉积中的作用。试验选取雄性成年泰迪型贵宾犬50只,适应性饲喂2周后分成对照组(A组,n=6)和高脂饲粮组(n=44)。对照组饲喂基础饲粮,高脂饲粮组饲喂高脂饲粮(基础饲粮+25%猪油)。饲喂8周后高脂饲粮组成功建立肥胖犬模型,筛选30只肥胖犬,等分成5组:高脂饲粮对照组(B组,n=6)、银杏叶提取物组(C组,n=6)、阴性siRNA (N-siRNA)对照组(D组,n=6)、APN-siRNA组(E组,n=6)和AdipoR1-siRNA组(F组,n=6)。A组继续饲喂基础饲粮,其他各组继续饲喂高脂饲粮。C组、D组、E组和F组每天按50 mg/kg BW的剂量灌服银杏叶提取物,A组和B组灌服等量生理盐水。此外,D组、E组和F组在腹部皮下注射500 μL siRNA,A组、B组和C组注射等体积生理盐水,每周3次。试验周期为4周。试验结束后,测量试验犬的体重及体脂率,测定血清甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)及APN含量,苏木精-伊红(HE)染色观察脂肪细胞大小变化,分别用实时荧光定量PCR (qRT-PCR)法和Western blot法检测APN/AdipoR1/AMPK信号通路中相关分子基因表达和蛋白表达变化。结果显示:1) B组体重及体脂率显著高于A组与C组(P<0.05),C组体重及体脂率显著低于E组与F组(P<0.05)。2) B组血清TG、TC、LDL-C及APN含量显著高于A组与C组(P<0.05),HDL-C含量显著低于A组与C组(P<0.05);C组血清TG、TC、LDL-C含量显著低于E组和F组(P<0.05),HDL-C含量高于E组和F组(P<0.05)。3) C组和D组皮下脂肪组织中脂肪细胞面积显著小于B组、E组和F组(P<0.05)。4) C组脂肪组织中APN的mRNA和蛋白相对表达量显著高于B组与E组(P<0.05),AdipoR1的mRNA和蛋白相对表达量显著高于B组与F组(P<0.05),腺苷单磷酸活化蛋白激酶α1(AMPKα1)的mRNA相对表达量和磷酸化腺苷单磷酸活化蛋白激酶α1(p-AMPKα1)的蛋白相对表达量显著高于B组、E组与F组(P<0.05)。综上所述,银杏叶提取物可以降低肥胖犬的体重、体脂率和血脂水平,这与银杏叶提取物激活脂肪组织APN/AdipoR1/AMPK信号通路,改善脂肪细胞脂质代谢与脂肪沉积的作用有关。

本文引用格式

陆江 , 朱道仙 , 卢劲晔 , 刘莉 , 刘静 . 银杏叶提取物通过脂联素/脂联素受体1/腺苷酸激活蛋白激酶信号通路改善肥胖犬脂质代谢及脂肪沉积的研究[J]. 动物营养学报, 2022 , 34(5) : 3261 -3270 . DOI: 10.3969/j.issn.1006-267x.2022.05.052

Abstract

The aim of this study was to evaluate the role of adiponectin (APN)/adiponectin receptor 1 (AdipoR1)/adenosine monophosphate-activated protein kinase (AMPK) signaling pathway in Ginkgo biloba extract improving lipid metabolism and fat deposition of obese dogs induced by high-fat diet. Fifty male adult Teddy poodles were divided into control group (group A, n=6) and high-fat diet group after adaptive feeding for 2 weeks. Dogs in high-fat diet group were fed a high-fat diet, and those in control group were fed a basic diet. After 8 weeks, the obese dog model induced by high-fat diet was successfully established. Thirty obese dogs in high-fat diet group were screened and divided into 5 groups:high-fat diet control group (group B, n=6), Ginkgo biloba extract group (group C, n=6), negative siRNA control group (group D, n=6), APN-siRNA group (group E, n=6) and AdipoR1-siRNA group (group F, n=6). Dogs in control group continued to fed the basic diet, and those in other groups were continued to fed the high-fat diet. Dogs in groups C, D, E and F were administrated with 50 mg/kg BW Ginkgo biloba extract every day, while the groups A and B were administrated with equal amount of normal saline. In addition, dogs in groups D, E and F were immunosuppressed with injection of 500 μL siRNA three times a week, while the groups A, B and C were immunosuppressed with injection of equal volume of normal saline. The trial period was 4 weeks. After the experiment, the body weight, body fat rate, and the contents of serum triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C) and APN were measured, and hematoxylin-eosin (HE) staining was used to observe the changes of adipocytes size. The gene expression and protein expression changes of related molecules in APN/AdipoR1/AMPK signaling pathway were detected by real-time quantitative PCR (qRT-PCR) and Western blot method, respectively. The results showed as follows:1) the body weight and body fat rate in group B were significantly higher than those in groups A and C (P<0.05), while those in group C were significantly lower than those in groups E and F (P<0.05). 2) The contents of serum TG, TC, LDL-C and APN in group B were significantly higher than those in groups A and C (P<0.05), while the HDL-C content was significantly lower than that in groups A and C (P<0.05). The contents of serum TG, TC and LDL-C in group C were significantly lower than those in groups E and F (P<0.05), while the HDL-C content was significantly higher than that in groups E and F (P<0.05). 3) The area of adipocytes in subcutaneous adipose tissue in groups C and D was significantly smaller than that in groups B, E and F (P<0.05). 4) The APN mRNA and protein relative expression levels in subcutaneous adipose tissue in group C were significantly higher than those in groups B and E (P<0.05), the AdipoR1 mRNA and protein relative expression levels were significantly higher than those in groups B and F (P<0.05), and the AMPKα1 mRNA relative expression level and the phospho-AMPKα1 (p-AMPKα1) protein relative expression level in group C were significantly higher than those in groups B, E and F (P<0.05). In summary, Ginkgo biloba extract can reduce the body weight, body fat rate and blood lipid level of obese dogs, which is related to the effect of Ginkgo biloba extract on up-regulating APN/AdipoR1/AMPK signal pathway and improving adipocyte lipid metabolism and fat deposition in adipose tissue.

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