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Development in immunochemical control of regulatory enzymes and receptors concerned in the lipid metabolism and the application for low-fat meat production

脂质代谢相关调节酶和受体的免疫化学控制研究进展及其在低脂肉生产中的应用

基本信息

批准号：
10556060
负责人：
AKIBA Yukio
金额：
$ 7.3万
依托单位：
TOUHOKU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B).
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2000
项目状态：
已结题

项目摘要

The present study was conducted to identify enzymes and receptors involved in fat accumulation and to control by immunochemical methods to reduce obesity domestic animals and consequently produce "Low fat meat".1. Adipose tissue lipoprotein lipase (LPL)We could have obtained 24 positive clones of hybridoma to bind specifically with chicken LPL and 3 clones out of 24 were effective to reduce LPL activity in adipose tissues. In chickens received continuous infusion of LPL antibody (CLP-16) for 14 days, adipose fat pad weight was significantly lower than the control chickens whereas the body weight decreased to a small extent. These findings suggest that persistent administration of anti-LPL monoclonal antibody is effective mean to modify LPL activity and consequently manipulate fatness in broiler chickens.2. Adipose tissue lipoprotein receptor (LR)It was shown following western blot analysis using antiserum of various LRs (LRP, LRP380, and LR8) that LR8 was present in chicken adipose tissues and concerned in the chicken adiposity. However, administration of LR8 antibody did not modify VLDL uptake to chicken adipocytes.3. Hepatic lipoprotein receptor (HLR)It was not able to identify the new cDNA clone of chicken HLR though the screening of chicken liver cDNA library. However, new LDL binding proteins (100 and 130kDa) were detected in the ligand blot analysis in chicken liver membrane. The new LDL binding proteins were characterized by no-binding Ca ion and receptor associated protein (RAP), which were different from LDL receptor family obtained in mammals.4. Plasma cholesteryl ester transfer protein (CETP)The CETP activity in chicken plasma was high before laying eggs whereas that in laying hens was not detectable. These results suggested that chicken CETP play an important role to the cholesterol metabolism. We have been trying to obtain chicken CETP cDNA clone using human CETP probe, while at present chicken CETP cDNA sequence is still not available.

本研究旨在鉴定与脂肪积累有关的酶和受体，并通过免疫化学方法控制，以减少家畜肥胖，从而生产“低脂肪肉”。脂肪组织脂蛋白脂肪酶（LPL）我们获得了24个与鸡LPL特异性结合的阳性杂交瘤克隆，其中3个克隆能有效降低脂肪组织中LPL的活性。连续注射LPL抗体（CLP-16） 14 d后，脂肪垫重显著低于对照组，体重略有下降。上述结果表明，持续给药抗LPL单克隆抗体是改变肉鸡LPL活性，从而控制肉鸡肥胖的有效手段。脂肪组织脂蛋白受体（LRP， LRP380和LR8）的抗血清western blot分析表明，LR8存在于鸡脂肪组织中，与鸡肥胖有关。然而，LR8抗体并没有改变VLDL对鸡脂肪细胞的吸收。肝脂蛋白受体（liver lipoprotein receptor， HLR）通过鸡肝脏cDNA文库的筛选，未能鉴定出新的HLR cDNA克隆。而在鸡肝膜的配体印迹分析中，检测到新的LDL结合蛋白（100和130kDa）。新的LDL结合蛋白以不结合Ca离子和受体相关蛋白（RAP）为特征，与哺乳动物中获得的LDL受体家族不同。血浆胆固醇酯转移蛋白（CETP）鸡下蛋前血浆CETP活性较高，而蛋鸡未检测到CETP活性。以上结果提示，鸡CETP在胆固醇代谢中起重要作用。我们一直在尝试用人CETP探针获得鸡CETP cDNA克隆，但目前还没有鸡CETP cDNA序列。