IDENTIFICATION OF SPECIES DIFFERENCES IN LIPOPROTEIN LIPASE AND MANIPULATION OF FAT DEPOSITION BY MONOCLONAL ANTIBODY FOR THE LIPASE IN CHICKENS

鸡体内脂蛋白脂肪酶的物种差异鉴定及脂肪酶单克隆抗体对脂肪沉积的调控

• 批准号: 05454110
• 负责人: AKIBA Yukio
• 金额: $ 4.35万
• 依托单位: TOHOKU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05454110/
• 关键词: LIPOPROTEIN LIPASE; OBESITY; FAT DEPOSITION; SPECIES SPECIFICITY; MONOCLONAL ANTIBODY; CHICKEN; LIPOPROTEIN BREAKDOWN; 脂質代謝

Obesity (excessive fat deposition) is a metabolic disorder common in human and animals. Lipoprotein lipase (LPLase) in adipose tissues plays a major role in determining fat deposition in chickens. Present study was undertaken to identify species differences in LPLase in chickens and mammals and to explore the possible ways to manipulate the excess fat deposition by administration of the monoclonal antibodies of LPLase.(1) Hydolysis of lipoproteins by LPLase was markedly inhibited by Triton WR-1339 in rats but inhibited to a small extent in chickens, suggesting existence of species differences in the LPLase.(2) Glycerol was identified for an excellent reagent to stabilize LPLase activity, because LPLase was sensitive to the decomposition. LOLases of chickens and rats were purified with high specific activity by applying hydroxyapatite column.(3) Purified LPLase of chickens differed from that of rats in terms of Km, Vm and kinetics on inhibition by Triton WR-1339.(4) Ultrafiltration and Mono Q columns for apoproteins yielded a purified Apo C-II of chickens. Activation of LPLase by the Apo C-II was potent in chickens more than in rats.(5) Purified LPLase was immunologically challenged to mice to produce the hybridomas and 24 clones characterized with high affinity to LPLase were identified. Three monoclonal antibodies out of 24 clones demonstrated marked inhibitory activity on the lipoprotein hydrolysis by LPLase.(6) Intraperitoneal administration of the monoclonal antibody for LPLase resulted in an inhibition by 20% of adipose tissue LPLase activity and a marked increase in plasma triglyceride concentration in chickens, suggesting that the monoclonal antibody administration may be a possible procedure to retard excessive fat deposition in animals.

肥胖(脂肪过度沉积)是一种在人类和动物中常见的代谢紊乱。脂肪组织中的脂蛋白脂酶(LPLase)在决定鸡脂肪沉积方面起着重要作用。本研究旨在鉴定鸡和哺乳动物LPLase的物种差异，并探索通过注射LPLase的单抗来控制LPLase过度脂肪沉积的可能途径。(1)Triton WR-1339显著抑制LPLase对脂蛋白的水解，但对鸡的LPLase有轻微的抑制作用，提示LPLase存在物种差异。(2)甘油被确定为稳定LPLase活性的良好试剂，因为LPLase对分解敏感。(3)纯化的鸡和大鼠的LPLase在Km、Vm和Triton WR-1339抑制动力学方面与大鼠不同。(4)超滤柱和Mono Q柱分离纯化的载脂蛋白C-II。ApoC-II对LPLase的激活作用在鸡中比在大鼠中更强。(5)将纯化的LPLase免疫小鼠制备杂交瘤细胞，筛选出24个与LPLase具有高亲和力的克隆。24个克隆中有3个对LPLase的脂蛋白水解酶有明显的抑制活性。(6)经腹腔注射LPLase单抗后，鸡脂肪组织LPLase活性抑制20%，血浆甘油三酯浓度显著升高，提示单抗给药可能是一种延缓动物脂肪过度沉积的方法。

项目成果

Kazuaki Takahashi et al.: "Effect of Methionine Supplementation on Lipogenesis and Lipolysis in Broiler Chicks" Japanese Poultry Science. 32. 99-106 (1995)

Kazuaki Takahashi 等人：“补充蛋氨酸对肉鸡脂肪生成和脂肪分解的影响”日本家禽科学。

K.Sato et al.: "Species differences between chicks and rats in inhibition of lipoprotein hydrolysis by Triton WR-1339" Comparative Biochemistry Physiology. 112C. 315-319 (1995)

K.Sato 等人：“Triton WR-1339 抑制脂蛋白水解方面小鸡和大鼠之间的物种差异”比较生物化学生理学。

Yukio Akiba et al.: "Effect of Dietary Fat and Protein Sources on Performance,Lipid Content and Mixed Function Oxidase in Liver,and Fat Deposition and Adipocyte Cellularity in Abdomen in Broiler Chickens" Japanese Poultry Science. 31. 381-391 (1994)

Yukio Akiba 等人：“膳食脂肪和蛋白质来源对肉鸡生产性能、肝脏中脂质含量和混合功能氧化酶以及腹部脂肪沉积和脂肪细胞细胞性的影响”《日本家禽科学》。

Yukio Akiba et al.: "The Effects of dietary Lipid on Poultry Performance and Composition" Proc. Aust. Poultry Science Sym. 7. 1-8 (1995)

Yukio Akiba 等人：“日粮脂质对家禽性能和成分的影响”Proc。

Y.Akiba et al.: "Effect of dietary fat and protein sources on performance, lipid content and mixed function oxidase in liver, and fat deposition and adiposyte cellularity in abdomen in broiler chickens" Japanese Poultry Science. 31. 381-391 (1994)

Y.Akiba 等人：“日粮脂肪和蛋白质来源对肉鸡生产性能、脂质含量和肝脏混合功能氧化酶以及腹部脂肪沉积和脂肪细胞构成的影响”《日本家禽科学》。