CN115245510A - 反式-10，顺式-12共轭亚油酸的用途 - Google Patents
反式-10，顺式-12共轭亚油酸的用途 Download PDFInfo
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Abstract
本发明涉及反式‑10，顺式‑12共轭亚油酸异构体的用途，具体公开了反式‑10，顺式‑12共轭亚油酸在制备导致体重减轻、改善胰岛素敏感性、升高基础代谢率、缓解神经性厌食症、抗抑郁的产品中的用途。本发明还公开了一种包含共轭亚油酸的治疗肥胖症、糖尿病、神经性厌食症和抑郁症的药物组合物。本发明首次发现所述共轭亚油酸对棕色脂肪以及下丘脑相关基因的影响。
Description
技术领域
本发明涉及医药技术领域，具体涉及反式-10，顺式-12共轭亚油酸(trans-10，cis-12 conjugated linoleic acid，以下简称t10c12-CLA)在制备导致体重减轻、改善胰岛素敏感性、升高基础代谢率、缓解神经性厌食症、抗抑郁的产品中的用途。
背景技术
共轭亚油酸(CLA)是一类具有较高生理活性的脂肪酸，其中的顺式-10，反式-12共轭亚油酸，主要存在于牛、羊等反刍动物肉和奶中，但是量很少，以牛奶为例进行估算，一升牛奶中大概有0.14g左右，约占乳脂总量的0.35％。即使是长期的、食用较多牛羊肉奶的人，他们的脂肪和血浆中也检测不出t10c12-CLA(Castro-Webb N，Ruiz-Narvaez EA，CamposH.2012.Cross-sectional study of conjugated linoleic acid in adipose tissueand risk of diabetes.Am J Clin Nutr.96:175-81)。t10c12-CLA能够降低体脂和血脂，减少肥胖症的发生概率，独特的生理作用使其成为近年来研究的明星营养素。早在2008年，美国FDA就为t10c12-CLA等共轭亚油酸颁发了食品许可证。但不同于天然的肉奶CLA，目前市售的CLA主要为工业氢化CLA复合物，其中t10c12-CLA和c9t11-CLA含量基本相同，共占总量的80-95％，除此之外还有5-20％不等的杂质异构体，常标识成CLA复合物(50:50)，这些杂质在食用过程中是否具有健康风险，尚存疑问。
最初人们发现，t10c12-CLA的生物活性是减少了鼠类的白色脂肪(Park Y，Albright KJ，Liu W，Storkson JM，Cook ME，Pariza MW.1997.Effect of conjugatedlinoleic acid on body composition in mice.Lipids.32:853-58)、后来又发现它能下调鼠类和家畜的乳脂合成(Hussein M，Harvatine KH，Weerasinghe WM etal.2013.Conjugated linoleic acid-induced milk fat depression in lactatingewes is accompanied by reduced expression of mammary genes involved in lipidsynthesis.J Dairy Sci.96:3825-34)。此期间，有研究指出，t10c12-CLA可能会干扰体内相关的激素或信号分子，引起肝脏脂肪变性等，对机体产生一定的影响(Vyas D，KadegowdaAK，Erdman RA.2012.Dietary conjugated linoleic Acid and hepatic steatosis:species-specific effects on liver and adipose lipid metabolism and geneexpression.J Nutr Metab.2012:928-32)。另外，人类长期使用安全剂量的t10c12-CLA，既无减脂效果，也无副作用发生(Kim JH，Kim Y，Kim YJ&Park Y.2016.Conjugated LinoleicAcid:Potential Health Benefits as a Functional Food Ingredient.Annu Rev FoodSci Technol.7:10.1-10.24)。
更为关键的是，科学家探讨了t10c12-CLA与下丘脑控制中枢的关系，却没有发现t10c12-CLA影响下丘脑能量平衡中枢的证据，t10c12-CLA对下丘脑功能的影响机制不甚清楚，t10c12-CLA影响脂肪代谢的活性作用机制也尚未可知，因此影响了人们对t10c12-CLA的进一步研究和应用。
在生物工程技术的支撑下，利用重组微生物生产无杂质t10c12-CLA饲喂动物、甚至研制自身合成t10c12-CLA的转基因动物，是比较理想的研究t10c12-CLA活性的一种途径。瘤胃痤疮丙酸杆菌的PAI异构酶，能够将亚油酸催化成t10c12-CLA(Hornung E，KruegerC，Pernstich C，Gipmans M，Porzel A，FEussner I.2005.Production of(10E,12Z)-conjugated linoleic acid in yeast and tobacco seeds.BBA-Mol Cell BiolLipids.1738:105-14)。专利申请人团队发现，密码子优化的pai基因，能够在重组的乳酸球菌中表达，并高效合成t10c12-CLA(苟克勉，李世丽。产共轭亚油酸的食品级重组乳酸乳球菌及其构建方法与应用，专利号：ZL 2013 1 0066107.1；授权公告日：2014年11月05日，已失效)，用这种重组菌(饲料中t10c12-CLA的含量是0.9％)连续饲喂小鼠一个月后，它们的体重会显著下降，白色脂肪显著减少，连续饲喂2个月时，肝脏才有轻微的脂肪变性(Li SL，Ma SY，Xu BR，Fan ZY，Li MJ，Cao WG，Gou KM.2015.Effects of trans 10，cis 12-conjugated linoleic acid on mice are influenced by the dietary fat contentand the degree of murine obesity.Euro J Lipid Sci Technol.117:1908-18)。
专利申请人团队进一步证实，pai基因也能够在小鼠的培养细胞中异源生产t10c12-CLA(苟克勉，李世丽。亚油酸异构酶在脱氢和异构方面的双重用途，专利号：ZL2011 10112685.5；授权公告日：2013年3月20日，已失效)，故而，申请人利用动物转基因的方法，研制了一个转基因小鼠模型用于研究t10c12-CLA的活性作用机制。
发明内容
本发明的目的是提供共轭亚油酸，尤其是t10c12共轭亚油酸对于受试者能量代谢，尤其是对于棕色脂肪产热性能以及下丘脑应激相关的基因的影响。
为实现上述目的，本发明采取的技术方案为：
本发明一个方面提供了共轭亚油酸异构体在制备导致体重减轻、改善胰岛素敏感性、升高基础代谢率、缓解神经性厌食症、抗抑郁的产品中的用途。
本发明第二个方面提供了共轭亚油酸异构体在制备提高受试者棕色脂肪产热机能或提高受试者的维持净能的产品中的用途。
本发明第三个方面提供了共轭亚油酸在制备降低受试者白色脂肪细胞数量以及白色脂肪细胞体积的产品中的用途。
本发明第四个方面提供了共轭亚油酸异构体在制备调节下丘脑中能量消耗基因的产品中的用途，其中调节下丘脑中能量消耗基因选自下调orexin家族基因，或上调grp78基因。
本发明第五个方面提供了共轭亚油酸异构体在制备调节下丘脑中影响能量摄入基因的产品中的用途，其中调节下丘脑中影响能量摄入基因选自上调下丘脑细胞的ghrelin受体基因、上调下丘脑细胞中的胰岛素受体基因、下调厌食神经细胞POMC的标志基因cart、下调Agrp/POMC神经元下游的黑皮质素受体MC4R基因、下调食欲素家族基因。
在本发明的技术方案中，影响能量摄入的基因选自诱发受试者进食或厌食的基因。
本发明第六个方面提供了共轭亚油酸异构体在制备调节下丘脑中应激基因的产品中的用途，所述调节下丘脑中应激基因选自上调pai下丘脑细胞的糖皮质激素受体基因Nr3c1。
本发明第七个方面提供了共轭亚油酸异构体在制备促进棕色脂肪组织内ucp1基因、ppar-γ基因、prdm6基因、ucp2基因、ampk基因和ppar-δ基因表达的产品中的应用。
在本发明的技术方案中，所述共轭亚油酸异构体为t10c12共轭亚油酸，化学结构式为：
进一步地，所述共轭亚油酸异构体能够增强“下丘脑-垂体-肾上腺轴系”的负反馈调节。
进一步地，所述共轭亚油酸异构体能够干扰下丘脑中的miRNA调控机制。
进一步地，所述共轭亚油酸异构体能够改变棕色脂肪细胞的产热机制、增强产热性能。
进一步地，所述共轭亚油酸异构体能够减少白色脂肪组织。
进一步地，所述共轭亚油酸异构体通过口服、注射、外源基因异源表达的方式完成。
本发明第八个方面提供了一种包含共轭亚油酸异构体的治疗肥胖症、糖尿病、神经性厌食症和抑郁症的药物组合物。
进一步地，所述共轭亚油酸异构体为t10c12-CLA。
进一步地，所述t10c12-CLA作为药物组合物的唯一活性成分。
本发明具有如下优点或者有益效果：本发明提供了t10c12-CLA在制备导致体重减轻、改善胰岛素敏感性、升高基础代谢率、缓解神经性厌食症、抗抑郁的药物和保健品中的应用。本发明还提供了一种包含共轭亚油酸的治疗肥胖症、糖尿病、神经性厌食症和抑郁症的药物组合物。本发明通过建立pai小鼠模型，证实t10c12-CLA能够减少小鼠的白色脂肪组织，并首次发现t10c12-CLA改变棕色脂肪细胞的产热机制，通过激活小鼠的产热通路，增强产热机能，使其维持净能升高，导致白色脂肪组织减少；并进一步发现t10c12-CLA能够作用于下丘脑的多个功能基因，影响了进食、能量动态平衡、能量消耗、应激等过程；本发明为t10c12-CLA影响棕色脂肪细胞和下丘脑功能的相关性应用提供了依据和方法。
附图说明
图1是实施例1中同源重组载体pRosa-pai的结构示意图(A)、酶切鉴定图谱(B)以及打靶载体pRosa-Crispr结构示意图(C)。
图2是实施例1中pai小鼠的外源基因分析图(用PCR(A)和Southern blot(B)方法鉴定pai小鼠，图中pai，wt和Positive分别为pai小鼠、野生型小鼠和阳性对照的DNA样品；用RT-PCR(C)和Real-time PCR(D)方法分析pai小鼠组织中外源基因的表达，图中内参基因采用gapdh；用Western blot(E)方法鉴定pai小鼠组织中的PAI蛋白(48.9kDa)。
图3是pai(A)和野生型(B)小鼠胎儿组织的高压气相色谱部分图解。
图4是野生型(左)和pai小鼠(右)腹腔白色脂肪的核磁扫描(上)及组织学(下)分析示意图。
图5是pai小鼠和野生型小鼠的代谢参数。
图6是pai小鼠和野生型小鼠棕色脂肪细胞中产热基因的Real-time PCR分析。
图7是pai小鼠和野生型小鼠下丘脑功能基因的Real-time PCR分析。
具体实施方式
下述实施例仅仅是本发明的一部分实施例，而不是全部的实施例。因此，以下提供的本发明实施例中的详细描述并非旨在限制要求保护的本发明的范围，而是仅仅表示本发明的选定实施例。基于本发明的实施例，本领域技术人员在没有作出创造性劳动的前提下所获得的所有其他实施例，都属于本发明的保护范围。
本发明实施例中所使用的t10c12-CLA指的是反式-10，顺式-12共轭亚油酸，化学结构式为：
在本发明中，所述“维持净能”指维持人体或动物体基础生理活动所述的能量，所述的基础生理活动包括呼吸、血液循环、离子浓度调节、肌肉收缩、代谢废物的排出、受损组织的修复、蛋白质周转、酶的合成、维持体温以及其他维持生命活动必须的能量。
实施例1：pai小鼠的构建
实验步骤
(1)同源重组载体pRosa-pai的构建
将专利发明人以前构建的含有优化的pai基因序列的pPAI-IRES-AcGFP质粒(苟克勉，李世丽，亚油酸异构酶在脱氢和异构方面的双重用途，专利申请号：201110112685.5)PCR扩增至pTurbo-Cre(GenBank accession no.AF334827)，构成原始的pCAGGS-pai质粒；随后在CAG启动子前面插入Pac I和Spe I酶切位点；然后在polyA后面插入Mfe I、Asc I、Pst I和Hind III酶切位点；经过两个中间质粒后，再将pai基因片段用Xba I和Bgl II酶切位点导入第二个中间质粒，构成pCAG-pai质粒；最后，再将这个质粒中的表达元件CAG-pai-polyA，用Pac I和Asc I酶切位点插入pRosa26质粒(Addgene#21271)中，获得同源重组质粒pRosa-pai(13.53kb)，其中左右同源臂分别长约1.08kb和4.3kb，pai基因的高效表达元件长约3.4kb(图1A)，最后，多种酶切和测序证明载体构建正确(图1B)。
(2)打靶载体pRosa-Crispr载体的构建
将来自pEF.myc.ER-E2-Crimson质粒(Addgene#38770)的EF1a启动子，通过Kpn I和Nco I双酶切置换到pDG330质粒(Addgene#100898)，此中间质粒的Cas9表达元件上下游各有一个U6-sgRNA-sgRNA scaffold的表达结构，最后，用标准的Bbs I酶切，将2个相同的sgRNAs分子(5-act cca gtc ttt cta gaa ga-3)插入到U6启动子后面，完成了pRosa-CRISPR打靶载体的构建(图1C)，这2个sgRNAs可以有效地将Cas9蛋白酶引导到Rosa26的Exon-1和Exon-2间特定序列，完成精准的基因组DNA的切割过程。
(3)显微注射
上述pRosa-pai和pRosa-Crispr质粒，分别经酶切和测序鉴定后，参照标准方法和试剂盒说明书，生产显微注射用DNA。注射前，Pvu I+Sal I酶切回收和纯化的pRosa-pai线性化重组载体(9203bp)，终浓度50～100ng/μl，与终浓度10ng/μl的pRosa-CRISPR质粒以2:1比例混匀后，参照标准的胚胎操作和转基因方法，获取C57BL/6小鼠受精卵，并进行原核的显微注射。
(4)转基因小鼠的分子鉴定
取三周龄小鼠的尾巴组织，用标准的SNET+蛋白酶K溶液裂解后，用标准的苯酚/氯仿/异戊醇(25:24:1)抽提DNA，用于PCR和Southern blot鉴定。
PCR分析发现(Forward:5-TAA CCA TGT TCA TGC CTT CTT C-3，Reverse:5-CACCTT GTT GTA GTG TCC GTT T-3)，pai小鼠基因组中有514-bp的pai基因片段，并且其Rosa序列(F:5-GGA GTG TTG CAA TAC CTT TCT GGG AGT TC-3 R:5-TGT CCC TCC AAT TTT ACACCT GTT CAA TTC-3；217bp)被切断，无法扩增到片段(图2A)，显示pai基因表达元件准确地整合在小鼠的Rosa26座位上(图2A)。Southern blot(图2B)和测序也证明了这一点。
RT-PCR(图2C)和Real-time PCR(图2D)分析证实，整合在Rosa26座位的pai基因，能够在转基因小鼠的所有组织细胞中表达，表达量因组织而异。结果显示，pai基因在转基因小鼠的所有组织(心He，肝Li，脾S，肺Lu，肾K，肌肉Mu，脑B，下丘脑Hy，卵巢O，睾丸T，棕色脂肪BAT和白色脂肪WAT)中正常表达。用His-tag标签抗体(pai基因3-末端的标签)进行免疫印迹分析证实，随机选择的pai小鼠心脏、肝脏、脾脏和肾脏组织中，都能够检测到分子量为48.9kDa的PAI蛋白(图2E)。
(5)脂肪酸组分的高压气相色谱分析
收集18.5天的pai胎儿和野生型胎儿样品，取0.5克左右匀浆收集至带聚四氟乙烯瓶盖的玻璃甲基化管中，超声波破碎后，参照文献(陈青，柳青，吴至芳，王宗义，苟克勉.棉花脂肪酸脱氢酶FAD2能够实现转基因小鼠n-6脂肪酸的内源性生物合成.中国科学C辑：生命科学.2009；39(8):755-780)所述方法，进行高压气相色谱分析。具体的，每管加4毫升氯乙酰甲醇(氯乙酰:甲醇＝1:10)和2毫升正己烷，盖紧管盖，80℃水浴2小时，取出冷却至室温后，再加7％的K2CO3溶液5毫升，振荡混匀后静置10分钟，离心(1000rpm，5min)分离的上清直接进样到HP-88型色谱柱(美国Agilent公司)，使用HP7890全自动气相色谱仪(Agilent)进行分析。参照脂肪酸标准品(Sigma 47885-U和O5632)的峰面积，将仪器检测到的信号，用GC ChemStation Software(Agilent)软件处理，计算相对含量。结果发现转基因胎儿组织有t10c12-CLA，占总脂肪酸的0.3％，野生型胎儿组织中没有相应的色谱信号(图3)，表明pai小鼠体细胞能合成t10c12-CLA。
受t10c12-CLA的影响，转基因胎儿组织中，个别多不饱和脂肪酸的含量发生了改变，如亚油酸C18:2n6和花生四烯酸C20:4n6都显著(p<0.05)增加；而油酸的含量显著(p<0.05)下降。值得一提的是，t10c12-CLA 的存在还促进了DHA成分的积累(p<0.05)，而DHA对胎儿的神经系统和眼睛发育极为重要。
实施例2、pai小鼠白色脂肪研究实验
跟踪pai公鼠体重发现，它们的成年体重与同窝的野生型公鼠没有区别，但是，核磁扫描发现其腹腔白色脂肪减少(图4上)，解剖后称重，pai小鼠附睾白色脂肪也是减少的(1.29±0.42vs 1.67±0.45，p<0.05)。按照标准方法对白色脂肪组织用4％多聚甲醛4℃过夜固定；用60％到100％乙醇的梯度脱水(5％酒精梯度、每梯度20min，各两次)，然后二甲苯透明10min，重复2次后浸蜡；蜡I→蜡II→蜡III各1h后包蜡；常规的石蜡切片、烤片、二甲苯组织脱蜡、苏木精/伊红染色、封片等技术细节处理样品，随后用显微镜在20倍物镜下观察，发现转基因白色脂肪细胞的体积，明显小于野生型(图4下)。
标准方法裂解提取白色脂肪的RNAs后，DNase I消化和反转录获得cDNA，按照产品说明书，使用ChamQ SYBR qPCR Master Mix试剂盒(南京诺唯赞生物科技有限公司)，采用
GreenI嵌合荧光法，对leptin基因进行Real-time PCR检测，以持家基因gapdh为内参，结果显示pai小鼠白色脂肪细胞的leptin基因表达量只有野生型细胞的35％，进一步证实pai小鼠的白色脂肪组织是减少的。
实施例3、pai小鼠产热机制变化研究实验
利用22℃恒温的小鼠代谢实时系统(TSE Phenomaster)，每隔39分钟自动采集一次数据，连续监测72小时后，进行数据汇算，结果发现，在体重增长(图5A)、食物摄入量(图5B)、呼吸代谢率都正常的情况下，pai小鼠的基础运动量减少(图5C)、夜间饮水量(图5D)、O2消耗量(图5E)、CO2产量(图5F)、热量释放(图5G,5H)却显著增加(p<0.05)，显示t10c12-CLA导致转基因小鼠的维持净能升高。
根据产热增加的变化特点，分析主导产热的棕色脂肪发现，成年pai公鼠的棕色脂肪组织比野生型增重了35％(0.54±0.18vs 0.40±0.08，p<0.05)；Real-time PCR结果显示，pai小鼠的棕色脂肪细胞中，与产热相关的几个关键基因，如ucp1、ppar-γ、prdm6、ucp2、ampk、ppar-δ都是上调的(图6)，意味着，t10c12-CLA向上调整了pai小鼠的产热机能，这是小鼠维持净能升高的原因；同时，pai小鼠的能量消耗加大，也可能是白色脂肪被动减少的主要原因。
实施例4、pai小鼠下丘脑功能基因表达研究
下丘脑是一个非常重要的神经内分泌器官，参与能量动态平衡(包括进食和厌食)、能量消耗、生殖机能、青春期出现、昼夜节律和应激反应的调节等。
利用Real-time PCR对pai小鼠下丘脑的功能基因分析发现，pai小鼠下丘脑的17个功能基因中有9个基因是正常表达的，包括弓状核食欲促进型AGRP神经细胞群的标志基因npy(neuropeptide Y)和agrp(agouti-related peptide)、介导NPY信号的神经突触后Y1和Y5的受体Y1/Y5R、食欲抑制型POMC神经细胞的标志基因pomc、瘦素受体lepr及其下游信号通路上的关键因子stat3和ampk、促性腺激素释放激素基因gnrh和印迹基因H19等(图7)。
表达量发生显著变化的基因有8个，其中对主导能量消耗的基因分析发现：与棕色脂肪细胞产热密切相关的orexin1受体基因显著下调，该基因可以直接通过交感神经将信号传导到褐色脂肪组织，引起棕色脂肪的产热反应和能量消耗、白色脂肪组织米色化过程。另外，细胞响应刺激的主要内质网伴侣分子grp78基因显著上调，说明下丘脑主导的棕色脂肪燃烧，直接受到了t10c12-CLA的影响。
对主导能量摄入的基因分析发现：一般的，动物血中的瘦素leptin、胰岛素、胃饥饿素ghrelin、葡萄糖、氨基酸等因子是调控进食与否的关键，这些因子与下丘脑中对应的受体结合后，经过下丘脑/脑干回路等综合调控，会引起动物的进食或者厌食反应。对pai小鼠来说，除leptin受体(及其下游的stat3和ampk)正常外，下丘脑细胞的ghrelin受体和胰岛素受体基因都出现上调；另外，厌食神经细胞POMC的标志基因cart下调，意味着动物厌食反应减轻；位于Agrp/POMC神经元下游的黑皮质素受体MC4R下调，意味着中央黑皮质素系统激活自主神经系统和较高级大脑结构的能力减弱，因而对厌食行为的影响降低；促进食的食欲素家族基因orexin1下调等，正常情况下，这个基因受leptin和血糖的抑制，可以被Ghrelin激活，由于这些血液因子没有变化，暗示orexin1的下调是直接由t10c12-CLA引起的。所有上述这些基因的变化，充分说明下丘脑的进食和厌食机制，即能量动态平衡调控的多个基因、直接受到了t10c12-CLA的影响。
与应激相关的基因：”下丘脑-垂体-肾上腺轴系”(HPA轴)是应激的关键轴系，pai下丘脑细胞的糖皮质激素受体基因Nr3c1出现上调，该受体是介导肾上腺糖皮质激素的关键，此受体基因上调，可直接抑制POMC神经元，影响厌食行为；更为关键的是，这意味着糖皮质醇反馈调节的能力改变，小鼠的应激反应增强，继而增强HPA轴的负反馈调节发挥抗抑郁效应，也说明下丘脑主导的应激反应，直接受到了t10c12-CLA的影响。
与microRNA调控的相关基因：如RNA修饰酶Dicer上调，它是形成microRNAs的关键基因，而下丘脑细胞中功能基因的microRNAs转录调控广泛存在，可以说，microRNAs介导的转录调控机制，几乎存在于下丘脑主导的所有生理功能中。由此看见，t10c12-CLA直接干扰了下丘脑中常见的miRNA调控机制，因此可以将t10c12-CLA应用到下丘脑的诸如进食、能量平衡控制、能量消耗、生殖、免疫、应激等所有生理功能中。
以上所述仅为本发明的优选实施例，并非因此限制本发明的专利范围，凡是利用本发明说明书及附图内容所作的等效变换，或直接或间接运用在其他相关的技术领域，均同理包括在本发明的专利保护范围内。
Claims (10)
1.共轭亚油酸异构体在制备导致体重减轻、改善胰岛素敏感性、升高基础代谢率、缓解神经性厌食症、抗抑郁的产品中的用途。
2.共轭亚油酸异构体在制备提高受试者棕色脂肪产热机能或提高受试者的维持净能的产品中的用途。
3.共轭亚油酸异构体在制备调节下丘脑中能量消耗基因的产品中的用途，其特征在于，所述调节下丘脑中能量消耗基因选自下调orexin家族基因，或上调grp78基因。
4.共轭亚油酸异构体在制备调节下丘脑中影响能量摄入基因的产品中的用途，其特征在于，所述调节下丘脑中影响能量摄入基因选自上调下丘脑细胞的ghrelin受体基因、上调下丘脑细胞中的胰岛素受体基因、下调厌食神经细胞POMC的标志基因cart、下调Agrp/POMC神经元下游的黑皮质素受体MC4R基因、下调食欲素家族基因；所述影响能量摄入的基因选自诱发受试者进食或厌食的基因。
5.共轭亚油酸异构体在制备调节下丘脑中应激基因的产品中的用途，其特征在于，所述调节下丘脑中应激基因为上调下丘脑细胞的糖皮质激素受体基因Nr3c1。
6.共轭亚油酸异构体在制备上调棕色脂肪组织内ucp1基因、ppar-γ基因、prdm6基因、ucp2基因、ampk基因和ppar-δ基因的产品中的用途。
7.根据权利要求1-6任一所述的用途，其特征在于，所述共轭亚油酸异构体为反式-10，顺式-12共轭亚油酸，化学结构式为
8.一种包含共轭亚油酸异构体的治疗肥胖症、糖尿病、神经性厌食症和抑郁症的药物组合物。
9.根据权利要求8所述的药物组合物，其特征在于，所述共轭亚油酸异构体为反式-10，顺式-12共轭亚油酸。
10.根据权利要求8所述的药物组合物，其特征在于，所述共轭亚油酸异构体作为药物组合物的唯一活性成分。
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