长期高脂饮食导致的大鼠肥胖与小肠脂肪吸收相关分子的表达
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摘要
目的探讨高脂膳食诱导的肥胖大鼠小肠脂肪吸收相关分子apoB48、apoAIV及MTP表达水平的变化情况。方法选取出生21d,断乳3d的健康雄性SD大鼠60只,适应性喂养3d后,随机分为正常对照组20只[喂食基础饲料(290千卡/100g)]和高脂膳食组40只[喂食高脂饲料(430千卡/100g)]。大鼠均自由摄食和饮水,每周测量体重及身长。饲养24周后,高脂膳食组中肥胖大鼠共32只,随机选取20只作为肥胖组大鼠。测定记录大鼠体重,计算Lee’s指数,测定并记录大鼠腹腔脂肪重量。记录大鼠每天摄食量,采用滴定法测定大鼠粪便脂肪含量,计算小肠脂肪吸收率。采用比色法测定大鼠空腹血糖及血脂水平,放免法测定胰岛素水平,计算HOMA指数。RT-PCR及Real-time PCR法测定大鼠小肠apoB、apoAIV及MTP mRNA表达水平。蛋白质免疫印迹法测定大鼠小肠MTP蛋白表达水平。用ELISA法测定大鼠小肠匀浆液apoB48含量。结果肥胖组大鼠的体重、Lee's指数、腹腔脂肪湿重、空腹血糖、TC、TG、胰岛素水平、HOMA指数及小肠脂肪吸收率较正常对照组大鼠显著增高(P<0.05);RT-PCR及Real-time PCR结果均显示,与正常对照组比较,肥胖组大鼠小肠apoB、apoAIV及MTP的mRNA表达水平均显著升高(P<0.05);蛋白质免疫印迹结果显示,与正常对照组比较,肥胖组大鼠小肠MTP蛋白表达水平显著升高(P<0.01);与正常对照组比较,肥胖组大鼠小肠匀浆液apoB48含量显著升高(P<0.01)。结论高脂膳食诱导的肥胖大鼠小肠apoB48、apoAIV及MTP表达水平均显著增加,小肠脂肪吸收功能增强。
Objective To investigate the expression changes of intestinal fat absorption associated apoB48,apoAIV and MTP in high-fat diet induced obesity rats.Methods 21-day-old,weaning for 3 days healthy male Sprague-Dawley rats were randomized into normal control(n=20)and high fat diet group(n=40),fed with standard chow(290 kcal/100 g)and high-fat how(430 kcal/100 g)for 24 weeks,respectively.Food and water were supplied ad libitum and rats were housed in independently ventilated animal cages on a 12:12-h light:dark schedule and kept at 20-25°C.Weight and body and tail lengths were measured weekly during the next 24 weeks feeding.Mean body weights,Lee's indexes and abdominal fat were measured.Dietary and fetal fat were measured.Intestinal fat absorption ratio was calculated.Fasting glucose and lipids in the peripheral circulation were measured by enzymatic colorimetric method.Fasting serum insulin was measured by radioimmunoassay method to calcluded homeostatic model assessment(HOMA)value.Intestinal apoB48,apoAIV and MTP mRNA expression levels were determined by RT-PCR and Real-time PCR.Intestinal MTP protein expression levels were determined by Western blot assay.ApoB48 levels in the small intestine homogenate were determined by ELISA.Results Rats with high-fat diet induced obesity showed higher body weight,Lee's index,abdominal fat and intestinal fat absorption ratio,along with higher FPG,TG,TC,fasting serum insulin and HOMA value(P<0.05).RT-PCR and Real-time PCR results showed that rats in the obese group had a higher levels of apoB,apoAIV and MTP mRNA than rats in control group(P<0.01).Western blot results showed that MTP protein expression was enhanced in small intestine of obese group than control group(P<0.01).Conclusion High-fat diet could induce overproduction of apoB48,MTP and apoAIV in the intestine and enhanced absorption of fat.
Objective To investigate the expression changes of intestinal fat absorption associated apoB48,apoAIV and MTP in high-fat diet induced obesity rats.Methods 21-day-old,weaning for 3 days healthy male Sprague-Dawley rats were randomized into normal control(n=20)and high fat diet group(n=40),fed with standard chow(290 kcal/100 g)and high-fat how(430 kcal/100 g)for 24 weeks,respectively.Food and water were supplied ad libitum and rats were housed in independently ventilated animal cages on a 12:12-h light:dark schedule and kept at 20-25°C.Weight and body and tail lengths were measured weekly during the next 24 weeks feeding.Mean body weights,Lee's indexes and abdominal fat were measured.Dietary and fetal fat were measured.Intestinal fat absorption ratio was calculated.Fasting glucose and lipids in the peripheral circulation were measured by enzymatic colorimetric method.Fasting serum insulin was measured by radioimmunoassay method to calcluded homeostatic model assessment(HOMA)value.Intestinal apoB48,apoAIV and MTP mRNA expression levels were determined by RT-PCR and Real-time PCR.Intestinal MTP protein expression levels were determined by Western blot assay.ApoB48 levels in the small intestine homogenate were determined by ELISA.Results Rats with high-fat diet induced obesity showed higher body weight,Lee's index,abdominal fat and intestinal fat absorption ratio,along with higher FPG,TG,TC,fasting serum insulin and HOMA value(P<0.05).RT-PCR and Real-time PCR results showed that rats in the obese group had a higher levels of apoB,apoAIV and MTP mRNA than rats in control group(P<0.01).Western blot results showed that MTP protein expression was enhanced in small intestine of obese group than control group(P<0.01).Conclusion High-fat diet could induce overproduction of apoB48,MTP and apoAIV in the intestine and enhanced absorption of fat.
引文
[1] Dominiczak MH,Caslake MJ.Apolipoproteins:metabolic roleand clinical biochemistry applications[J].Ann Clin Biochem,2011,48(6):498-515.
[2]陈兴国,李庆新,李东阳,等.高脂诱导肥胖大鼠血清游离脂肪酸与血糖代谢的相关性[J].中国老年学杂志,2016,36(23):5810-5811,5812.
[3] Bois MC,Dasari S,Mills JR,et al.Apolipoprotein A-IV-Asso-ciated Cardiac Amyloidosis[J].J Am Coll Cardiol,2017,69(17):2248-2249.
[4]杨江涛,韩淑芬,焦俊,等.膳食纤维对高脂高胆固醇膳食诱导脂代谢紊乱小鼠实验的研究[J].医药前沿,2015,(20):51-52,53.
[5] Huang W,Liu R,Ou Y,et al.Octreotide promotes weight lossvia suppression of intestinal MTP and apoB48expression in diet-induced obesity rats[J].Nutrition,2013,29(10):1259-1265.
[6] Nakajima K,Nagamine T,Fujita MQ,et al.Apolipoprotein B-48:a unique marker of chylomicron metabolism[J].Adv ClinChem,2014,64(1):117-177.
[6] Farias G,Netto BDM,Bettini SC,et al.Neuroendocrine regulationof energy balance:Implications on the development and surgicaltreatment of obesity[J].Nutr Health,2017,23(3):131-146.
[7] Hernández Vallejo SJ,Alqub M,Luquet S,et al.Short-termadaptation of postprandial lipoprotein secretion and intestinalgene expression to a high-fat diet[J].Am J Physiol GastrointestLiver Physiol,2009,296(4):782-792.
[8] Tokuhara D,Nochi T,Matsumura A,et al.Specific expressionof apolipoprotein A-IV in the follicle-associated epithelium of thesmall intestine[J].Dig Dis Sci,2014,59(11):2682-2692.
[9]李鸿雁,颜克松,战大伟,等.常规饲料添加蛋黄和花生米的方法建立大鼠肥胖模型[J].实验动物科学,2015,32(3):48-50,61.
[10]黄微,刘瑞,郭韡,等.高脂膳食诱导的大鼠肥胖对小肠黏膜糖吸收功能的影响[J].卫生研究,2012,41(6):878-882.
[11] Guan M,Qu L,Tan W,et al.Hepatocyte nuclear factor-4al-pha regulates liver triglyceride metabolism in part through secre-ted phospholipase A GXIIB[J].Hepatology,2011,53(2):458-466.
[12]赵飞,郭庆辉.鱼油摄入对肥胖大鼠脂肪组织、瘦素表达和胰岛素水平的影响[J].营养学报,2013,35(2):146-149.
[13] Van de kamer jh,Ten bokkel huinink h,Weyers ha.Rapidmethod for thedetermination of fat in feces[J].J Biol Chem,1949,177(1):347-355.
[14] Rao R,Roche A,Febres G,et al.Circulating Apolipoprotein A-IV presurgical levels are associated with improvement in insulinsensitivity after Roux-en-Y gastric bypass surgery[J].SurgObes Relat Dis,2017,13(3):468-473.
[15] Nzekwu MM,Ball GD,Jetha MM,et al.Apolipoprotein B48:anovel marker of metabolic risk in overweight children[J].Bio-chem Soc Trans,2007,35(3):484-486.
[16] Wu J,Xiao Y,Liu J.et al.Potential role of ATM in hepatocyteendocytosis of ApoE-deficient,ApoB48-containing lipoprotein inApoE-deficient mice[J].International journal of molecular medi-cine,2014,33(2):462-468.
[17] Lee T L,Tsai Y H,Lin W J.et al.A New Differential P-Chan-nel Logic-Compatible Multiple-Time Programmable(MTP)Memory Cell With Self-Recovery Operation[J].IEEE ElectronDevice Letters,2011,32(5):587-589.
[18] Dalmau-CarolàJ.Erythema nodosum,a"red flag"during anti-TNF therapy[J].International journal of rheumatic diseases,2013,16(4):493-494.
[19] Guo W,Liu R,Qiang O,et al.Study of serum TNF-α,GTH andMDA levels in obesity-prone and obesity-resistant rats induced byhigh fat diet[J].Mod Prev Med,2009,36(1):2434-2437.
[20] Xiao C,Dash S,Morgantini C,et al.Intravenous Glucose A-cutely Stimulates Intestinal Lipoprotein Secretion in HealthyHumans[J].Arterioscler Thromb Vasc Biol,2016,36(7):1457-1463.
[21] Lee YS,Li P,Huh JY,et al.Inflammation is necessary forlong-term but not short-term high-fat diet-induced insulin resist-ance[J].Diabetes,2011,60(10):2474-2483.
[2]陈兴国,李庆新,李东阳,等.高脂诱导肥胖大鼠血清游离脂肪酸与血糖代谢的相关性[J].中国老年学杂志,2016,36(23):5810-5811,5812.
[3] Bois MC,Dasari S,Mills JR,et al.Apolipoprotein A-IV-Asso-ciated Cardiac Amyloidosis[J].J Am Coll Cardiol,2017,69(17):2248-2249.
[4]杨江涛,韩淑芬,焦俊,等.膳食纤维对高脂高胆固醇膳食诱导脂代谢紊乱小鼠实验的研究[J].医药前沿,2015,(20):51-52,53.
[5] Huang W,Liu R,Ou Y,et al.Octreotide promotes weight lossvia suppression of intestinal MTP and apoB48expression in diet-induced obesity rats[J].Nutrition,2013,29(10):1259-1265.
[6] Nakajima K,Nagamine T,Fujita MQ,et al.Apolipoprotein B-48:a unique marker of chylomicron metabolism[J].Adv ClinChem,2014,64(1):117-177.
[6] Farias G,Netto BDM,Bettini SC,et al.Neuroendocrine regulationof energy balance:Implications on the development and surgicaltreatment of obesity[J].Nutr Health,2017,23(3):131-146.
[7] Hernández Vallejo SJ,Alqub M,Luquet S,et al.Short-termadaptation of postprandial lipoprotein secretion and intestinalgene expression to a high-fat diet[J].Am J Physiol GastrointestLiver Physiol,2009,296(4):782-792.
[8] Tokuhara D,Nochi T,Matsumura A,et al.Specific expressionof apolipoprotein A-IV in the follicle-associated epithelium of thesmall intestine[J].Dig Dis Sci,2014,59(11):2682-2692.
[9]李鸿雁,颜克松,战大伟,等.常规饲料添加蛋黄和花生米的方法建立大鼠肥胖模型[J].实验动物科学,2015,32(3):48-50,61.
[10]黄微,刘瑞,郭韡,等.高脂膳食诱导的大鼠肥胖对小肠黏膜糖吸收功能的影响[J].卫生研究,2012,41(6):878-882.
[11] Guan M,Qu L,Tan W,et al.Hepatocyte nuclear factor-4al-pha regulates liver triglyceride metabolism in part through secre-ted phospholipase A GXIIB[J].Hepatology,2011,53(2):458-466.
[12]赵飞,郭庆辉.鱼油摄入对肥胖大鼠脂肪组织、瘦素表达和胰岛素水平的影响[J].营养学报,2013,35(2):146-149.
[13] Van de kamer jh,Ten bokkel huinink h,Weyers ha.Rapidmethod for thedetermination of fat in feces[J].J Biol Chem,1949,177(1):347-355.
[14] Rao R,Roche A,Febres G,et al.Circulating Apolipoprotein A-IV presurgical levels are associated with improvement in insulinsensitivity after Roux-en-Y gastric bypass surgery[J].SurgObes Relat Dis,2017,13(3):468-473.
[15] Nzekwu MM,Ball GD,Jetha MM,et al.Apolipoprotein B48:anovel marker of metabolic risk in overweight children[J].Bio-chem Soc Trans,2007,35(3):484-486.
[16] Wu J,Xiao Y,Liu J.et al.Potential role of ATM in hepatocyteendocytosis of ApoE-deficient,ApoB48-containing lipoprotein inApoE-deficient mice[J].International journal of molecular medi-cine,2014,33(2):462-468.
[17] Lee T L,Tsai Y H,Lin W J.et al.A New Differential P-Chan-nel Logic-Compatible Multiple-Time Programmable(MTP)Memory Cell With Self-Recovery Operation[J].IEEE ElectronDevice Letters,2011,32(5):587-589.
[18] Dalmau-CarolàJ.Erythema nodosum,a"red flag"during anti-TNF therapy[J].International journal of rheumatic diseases,2013,16(4):493-494.
[19] Guo W,Liu R,Qiang O,et al.Study of serum TNF-α,GTH andMDA levels in obesity-prone and obesity-resistant rats induced byhigh fat diet[J].Mod Prev Med,2009,36(1):2434-2437.
[20] Xiao C,Dash S,Morgantini C,et al.Intravenous Glucose A-cutely Stimulates Intestinal Lipoprotein Secretion in HealthyHumans[J].Arterioscler Thromb Vasc Biol,2016,36(7):1457-1463.
[21] Lee YS,Li P,Huh JY,et al.Inflammation is necessary forlong-term but not short-term high-fat diet-induced insulin resist-ance[J].Diabetes,2011,60(10):2474-2483.