CN113116818B - 包埋体及其在制备药物或食品中的用途 - Google Patents
包埋体及其在制备药物或食品中的用途 Download PDFInfo
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- CN113116818B CN113116818B CN201911364126.6A CN201911364126A CN113116818B CN 113116818 B CN113116818 B CN 113116818B CN 201911364126 A CN201911364126 A CN 201911364126A CN 113116818 B CN113116818 B CN 113116818B
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- lactalbumin
- capsaicin
- polypeptide
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Abstract
本发明提出了包埋体及其制备药物或食品中的用途,所述包埋体包括:α‑乳白蛋白多肽载体;以及辣椒素,所述辣椒素包埋于所述α‑乳白蛋白多肽载体中。本发明的包埋体可以有效地提高辣椒素被细胞摄取的效率,发挥降低脂肪生成的作用,具有广泛的应用前景。
Description
技术领域
本发明涉及食品药品领域,具体地,本发明涉及包埋体及其在制备药物或食品中的用途。
背景技术
辣椒素是辣椒的活性成分,它对哺乳动物包括人类都有刺激性并可在口腔中产生灼烧感。辣椒素和与其相关的化合物并称为辣椒元,是辣椒产生的次级代谢产物。纯辣椒素是一种斥水亲脂、无色无嗅的结晶或蜡状化合物。但是,由于辣椒素的水溶性差、具有刺激性辣味、半衰期短等自身缺陷,使其在应用上受到一定限制,例如减脂作用。
发明内容
本发明旨在至少在一定程度上解决现有技术中存在的技术问题至少之一。
在本发明的一个方面,本发明提出了一种包埋体。根据本发明的实施例,所述包埋体包括:α-乳白蛋白多肽载体;以及辣椒素,所述辣椒素包埋于所述α-乳白蛋白多肽载体中。
研究发现,α-乳白蛋白经酶解后的多肽片段中存在较显著的两亲性多肽片段。由此推测α-乳白蛋白酶解后的多肽可作为两亲性单体聚集形成胶束。由于辣椒素水溶性差、具有刺激性辣味、半衰期短,不易被细胞所摄取,从而难以发挥其功能。α-乳白蛋白多肽载体属于两亲性胶束,表面呈亲水性,内核呈疏水性,其具有可降解性且降解产物对生物体无毒、易调控、易与活性成分结合的特性。因此,将具有疏水性的辣椒素包埋于α-乳白蛋白多肽载体中,可以有效地提高辣椒素被细胞摄取的效率,保护其生物活性,掩盖或抵消刺激性辣味,延长半衰期,从而高效发挥其功能,尤其是降低脂肪的生成。另外,发明人惊奇地发现,单纯采用α-乳白蛋白多肽载体也具备一定的抑制脂肪生成的作用,因此,其可以与辣椒素协同发挥作用,增进抑制效果,使得该包埋体具有广泛的应用前景。
根据本发明的实施例,上述包埋体还可以具有下列附加技术特征:
根据本发明的实施例,所述包埋体的粒径大于所述α-乳白蛋白多肽载体的粒径,其中,所述包埋体和α-乳白蛋白多肽载体的粒径分别独立地为15~25纳米。在此粒径下的纳米载体体积较小,可以避免空间阻塞,更容易穿透肠道粘液,从而被肠上皮绒毛细胞吸收利用。然而,小于10nm的纳米载体,表面曲率较高,在吸附过程中限制了纳米载体与上皮细胞表面的相对取向,导致膜包裹率短,细胞吸收效率低。
根据本发明的实施例,基于所述α-乳白蛋白多肽载体的质量,所述辣椒素的质量为10~15质量%。由此,接近α-乳白蛋白多肽载体的最大包埋量,达到充分包埋辣椒素的效果。
根据本发明的实施例,所述包埋体是通过下列方式获得的:将α-乳白蛋白溶解于缓冲液中,再将所得到的混合液与蛋白水解酶进行第一混合处理,以便得到多肽溶液;将所述多肽溶液与辣椒素进行第二混合处理,以便得到所述包埋体;其中,所述蛋白水解酶与所述α-乳白蛋白的质量比为1:(40~80)。在此配比下,α-乳白蛋白将被蛋白水解酶部分酶解,形成两亲性多肽,达到临界胶束浓度后自组装形成纳米胶束,表面呈亲水性,内核呈疏水性。由此,在疏水作用下能够将疏水性的辣椒素包埋到纳米胶束的内核中,从而获得包埋体。
根据本发明的实施例,所述蛋白水解酶选自地衣芽孢杆菌蛋白酶。由此,可以将α-乳白蛋白水解为具有两亲性的多肽。
根据本发明的实施例,进行所述第二处理之后,将所得到的混合液进行过滤处理,收集截留物进行干燥处理,以便得到所述包埋体。由此,以便于除去多余的Tris-HCl及辣椒素,提高包埋体的纯度。
根据本发明的实施例,所述截留物的分子量不小于4KD。由此,可以截留住包埋体,以除去多余的Tris-HCl及辣椒素,提高包埋体的纯度。
根据本发明的实施例,所述α-乳白蛋白多肽载体的制备方法包括:将α-乳白蛋白溶解于缓冲液中,再将所得到的混合液与蛋白水解酶进行混合,过滤,收集残留物进行干燥,以便得到所述α-乳白蛋白多肽载体。在一些实施例中,截留物的分子量不小于4KD。
将在本发明的另一方面,本发明提出了前面所述的包埋体或其中所限定的α-乳白蛋白多肽载体在制备药物或食品或试剂盒中的用途。根据本发明的实施例,所述药物或食品或试剂盒用于抑制机体中的脂肪生成。研究发现,辣椒素具有抑制脂肪生成的作用,但是,因其水溶性差、具有刺激性辣味、半衰期短,不易被细胞所摄取,从而难以发挥其功能。α-乳白蛋白多肽载体属于两亲性胶束,可以将疏水性的辣椒素包埋其中,从而有效地提高辣椒素被细胞摄取的效率,保护其生物活性,延长半衰期,从而高效发挥其功能。并且,发明人惊奇地发现,单纯采用α-乳白蛋白多肽载体也具备一定的抑制脂肪生成的作用,因此,可以单独发挥作用或者与辣椒素协同发挥作用,增进降低脂肪生成的效果。
根据本发明的实施例,所述包埋体或α-乳白蛋白多肽载体用于增加成熟白色脂肪细胞中的线粒体数量及提高线粒体的活性。由此,以便促进机体产能,减少脂肪生成。
根据本发明的实施例,所述包埋体或α-乳白蛋白多肽载体不被耐药泵蛋白排出在成熟白色脂肪细胞之外。通常情况下,药物进入机体后,细胞膜上的耐药泵蛋白(p-gp)会将药物排出,导致药物无法进入细胞内,发挥药效。发明人发现,包埋体或α-乳白蛋白多肽载体可以克服耐药泵蛋白的外排作用,进入细胞,发挥抑制细胞脂肪生成的作用。
根据本发明的实施例,所述包埋体或α-乳白蛋白多肽载体用于抑制成熟白色脂肪细胞中脂肪的合成和聚集。
根据本发明的实施例,所述包埋体或α-乳白蛋白多肽载体用于抑制成熟白色脂肪细胞中PPARγ及cEBPα基因的表达,促进TRPV1基因的表达。发明人发现,包埋体或α-乳白蛋白多肽载体能够通过抑制PPARγ及cEBPα基因的表达,促进TRPV1基因的表达,从而抑制脂肪生成。
在本发明的又一方面,本发明提出了一种抑制剂。根据本发明的实施例,所述抑制剂用于抑制前面所述的包埋体或其中所限定的α-乳白蛋白多肽载体进入细胞,所述抑制剂可抑制小窝蛋白的活性、网格蛋白的活性、大胞饮作用或微丝微管作用。发明人发现,包埋体或α-乳白蛋白多肽载体是在小窝蛋白、网格蛋白、大胞饮作用或微丝微管作用的辅助下进入细胞内的,因此,通过抑制小窝蛋白的活性、网格蛋白的活性、大胞饮作用或微丝微管作用,可以抑制包埋体或α-乳白蛋白多肽载体进入细胞,从而无法抑制脂肪生成,可以达到增脂目的,满足特殊需求。
根据本发明的实施例,所述抑制剂选自染料木素、盐酸氯丙嗪、阿米洛利或诺考达唑。盐酸氯丙嗪可以抑制网格蛋白辅助的内吞,染料木素可以抑制小窝蛋白辅助的内吞,阿米洛利可以抑制大胞饮作用,诺考达唑可以抑制微丝微管作用,这四者均可以抑制包埋体或α-乳白蛋白多肽载体进入细胞内,无法达到抑制脂肪生成的目的。
本发明的附加方面和优点将在下面的描述中部分给出,部分将从下面的描述中变得明显,或通过本发明的实践了解到。
附图说明
本发明的上述和/或附加的方面和优点从结合下面附图对实施例的描述中将变得明显和容易理解,其中:
图1显示了根据本发明一个实施例的α-乳白蛋白纳米球载体及包埋体的电镜图;
图2显示了根据本发明一个实施例的纳米胶束克服耐药泵蛋白P-gp的外排作用分析示意图;
图3显示了根据本发明一个实施例的纳米胶束的细胞内吞机制分析示意图;
图4显示了根据本发明一个实施例的纳米胶束溶酶体逃逸分析示意图;
图5显示了根据本发明一个实施例的载辣椒素纳米胶束对脂肪分化过程中脂质聚集的影响分析示意图;
图6显示了根据本发明一个实施例的模拟胃肠中辣椒素释放量分析示意图;
图7显示了根据本发明一个实施例的载辣椒素纳米胶束载不同pH值下的释放量分析示意图;
图8显示了根据本发明一个实施例的纳米胶束在脂肪部位的黏附时间示意图;
图9显示了根据本发明一个实施例的成熟白色脂肪细胞中的线粒体量比较示意图;
图10显示了根据本发明一个实施例的各组药物在3T3-L1成熟白色脂肪细胞中线粒体活力的影响分析示意图;
图11显示了根据本发明一个实施例的纳米胶束在体内的减脂功效研究分析示意图;
图12显示了根据本发明一个实施例的载辣椒素纳米胶束对脂肪分化蛋白表达的调控示意图。
具体实施方式
下面将结合实施例对本发明的方案进行解释。本领域技术人员将会理解,下面的实施例仅用于说明本发明,而不应视为限定本发明的范围。实施例中未注明具体技术或条件的,按照本领域内的文献所描述的技术或条件或者按照产品说明书进行。所用试剂或仪器未注明生产厂商者,均为可以通过市购获得的常规产品。
实施例1
在该实施例中,按照下列方式制备包埋有辣椒素的α-乳白蛋白纳米球载体,简称α-乳白蛋白纳米胶束。
准确称取α-乳白蛋白6mg于离心管,加入6mL 75mM Tris-HCl缓冲液(pH=7.5),在涡旋振荡器上使其充分溶解后,加入蛋白水解酶(地衣芽孢杆菌蛋白酶),蛋白水解酶与α-乳白蛋白的质量比为1:70。50℃恒温水浴内,将其加热30min,稍加冷却,得到多肽溶液。在多肽溶液中加入辣椒素(20%α-乳白蛋白质量),充分混匀后,多肽自组装形成包埋辣椒素的。用3KD的透析袋透析除去多余的Tris-HCl及辣椒素,将截留物于冻干机中冻干后得到胶束粉末。
图1的左图为α-乳白蛋白多肽载体(SNS),粒径为20nm左右,右图为包埋有辣椒素的α-乳白蛋白纳米球载体(SNS-Cap),粒径变大,为22nm左右。
采用等温滴定量热仪对载药纳米胶束进行表征,明确了辣椒素是与两亲性多肽以疏水相互作用结合而被包埋进纳米胶束中,负载量为126.58mg/g。
实施例2
将3T3-L1细胞接种到6-well细胞培养板中,分别处理单独的耐药泵糖蛋白的脂溶性特定荧光底物罗丹明123(Rh123,称作Rh123组)、被α-乳白蛋白纳米胶束(含辣椒素)包埋的Rh123(称作M(Rh123)组)以及Rh123和耐药泵糖蛋白抑制剂维拉帕米(Verapamil,称作Rh123+Verapamil组)的混合物,其中Rh123浓度为0.5μg/mL,Verapamil的浓度为0.5μM。在给药后不同的时间点取细胞,通过流式细胞仪检测被细胞吸收的Rh123的荧光强度,来验证各组的Rh123内吞量。
结果如图2所示,3T3-L1细胞单独处理Rh123时,由于耐药泵的外排作用,被内吞入系细胞的Rh123随时间增加没有明显增加,而耐药泵被抑制剂抑制后,Rh123的内吞量显著增加。当Rh123被包埋在α-乳白蛋白纳米胶束内时,在没有耐药泵抑制剂处理时,仍有大量内吞,表明α-乳白蛋白纳米胶束可有效克服耐药泵的外排作用。
实施例3
在确保对细胞正常代谢无影响的前提下,选择安全浓度下的四种不同的抑制剂对细胞内吞机制进行考察。盐酸氯丙嗪(Chlorpromazine)用于抑制网格蛋白辅助的内吞,染料木素(Genistein)抑制小窝蛋白辅助的内吞,阿米洛利(Amiloride)抑制大胞饮作用,诺考达唑(Nocodazole)用于抑制微丝微管作用,其有效作用浓度经CCK-8毒性实验得出分别为1.36、9.05、8.28和8.64μg/mL。
如图3的流式细胞分析结果所示,将未加入抑制剂的细胞对载脂溶性荧光染料Cy3的α-乳白蛋白纳米胶束(SNS-Cy3,不含辣椒素)的摄取量作为对照组,即细胞正常状态下的值,将该值定为百分之百。由图可知,在加入安全浓度下的四种不同抑制剂后,细胞对纳米胶束的摄取量都有所减少,对3T3-L1细胞来说,经染料木素处理的细胞相对于其他三种药物来说荧光值最小,摄取量减少的最多,即说明对载辣椒素纳米胶束的摄取主要是通过小窝蛋白介导的辅助作用被细胞摄取,其他几种方式可能起到辅助摄取作用。
实施例4
首先制备交联Cy5染料的α-乳白蛋白纳米胶束(未包埋辣椒素,简称SE-Cy5),用于标记纳米胶束在细胞内的分布,3T3-L1细胞接种在玻璃底面的共聚焦小皿中,加入SE-Cy5,并在8h后用Lyso-Traker染料标记活细胞中的溶酶体,用DAPI蓝色荧光染料标记细胞核,通过共聚焦显微镜观察不用荧光标记的纳米胶束(SE-Cy5)、溶酶体(Lysome)和细胞核(DAPI),并观察将三种荧光重叠后(Merge图)溶酶体和纳米胶束的同定位情况。
通过CLSM观察了3T3-L1细胞对纳米胶束的吞噬情况。如图4所示,蓝色荧光是用DAPI标记细胞核;绿色荧光是用Lyso-Tracker标记的溶酶体;橙色荧光为纳米胶束表面交联的SE-Cy5染料,Merge代表重叠图,Lysome代表溶酶体,SE-Cy5代表交联Cy5染料的α-乳白蛋白纳米胶束,DAPI代表用来标记细胞核的染料。
纳米胶束已经进入细胞,8小时后,橙色荧光与绿色荧光重合量很少,两种颜色在共定位图片中均清晰可见,说明此时纳米胶束已逃出溶酶体,纳米胶束可以帮助药物逃出溶酶体以便在细胞中进一步发挥作用。
实施例5
肥胖中的白色脂肪是许多有害因素的主要来源(如游离脂肪酸、活性氧等)。相反对于臭名昭著白脂,棕色脂肪组织有其独特之处,通过代谢活性的产热棕色脂肪细胞增加身体能量消耗的能力。虽然棕色脂肪在成年人中分布较少,棕色脂肪细胞样细胞(米黄色细胞)已被鉴定存在于白色脂肪中,特别是在皮下白色脂肪中。因此如何将白色脂肪棕色化是现在脂质代谢研究中重点。
将3T3-L1细胞接种在6-well细胞培养板上,当细胞长满整个培养孔的时候,NC组每两天更换正常细胞培养基,PC组和其他给药组(SNS-Cap组、DMSO-Cap组、SNS组)每两天更换正常细胞培养基及给予MDI药物,给药组在PC组的基础上同时给予SNS-Cap或者溶解在DMSO中的Cap,或者SNS。在经过8天后,PC组和其他药物组在MDI的催化下,分化为成熟的白色脂肪细胞,有大量的脂肪积累,用BODIPY-FITC绿色荧光染料标记脂滴,在给药后被标记的脂滴数量有不同程度的减弱。
如图5所示,其中,NC组每两天更换正常细胞培养基,PC组和其他给药组均更换MDI药物,给药组同时给予SNS-Cap或溶解在DMSO中的Cap,或者SNS。在经过8天后,3T3-L1细胞在MDI药物诱导下成功的分化成成熟的白色脂肪细胞(PC),在脂肪分化过程中持续给药,在分化完成后,BODIPY-FITC将脂滴染色后观察到,载辣椒素纳米胶束显示出最明显的降低脂质生成的功效,其次是溶解于DSMO中的辣椒素,有趣的是纳米载体(不载辣椒素)本身也显示出一定的降脂功效。
实施例6
首先制备模拟胃液及模拟肠液:称取1g胃蛋白酶溶解在100mL超纯水中,用HCl调节pH至1.2(模拟胃液)。称取0.68g磷酸二氢钾溶解在90mL超纯水中,用0.5M NaOH溶液调节pH至6.8,再加入1g胰蛋白酶,溶解后定容至100mL(模拟肠液)。称取10mg包埋辣椒素的纳米胶束冻干粉于5mL模拟胃液中,振荡均匀后在37℃的摇床中孵育,分别于在不同时间点取样。2h后将样品取出,用0.5M氢氧化钠溶液将pH调节至6.8,按照上述方法加磷酸二氢钾和胰蛋白酶配制成模拟肠液,振荡均匀后继续在37℃的摇床中孵育,分别于不同时间点取样后,用乙腈将析出的辣椒素提取出来,通过UPLC测定辣椒素的浓度,算出辣椒素的累计释放率。
如图6所示,α-乳白蛋白在形成纳米胶束后,在胃液中的释放只有20%-30%左右,有70-80%的纳米胶束可成功的在肠内释放,提高辣椒素的吸收。
实施例7
首先制备不同pH的Tris-HCl缓冲溶液,称取10mg包埋辣椒素的纳米胶束冻干粉于5mL的缓冲溶液中,在不同时间点收集析出的辣椒素,并通过UPLC测量辣椒素的累计释放率,通过比较在模拟的体液环境(pH 7.4)、溶酶体环境(pH 5.5)以及弱酸的脂肪细胞环境(pH 6.5)的释放率,解释包埋辣椒素的纳米胶束在脂肪部位的释放程度。
如图7所示,载辣椒素纳米胶束在弱酸性环境下(pH 6.5)的释放大于在正常体液下(pH 7.4)的释放,表明在弱酸性的脂肪环境下释放更多,并且在溶酶体pH 5.5条件下也可以释放,证明可以逃逸溶酶体。
实施例8
将两只体重相似的高脂饲料诱导的肥胖小鼠分别在腹股沟白色脂肪的原位注射相同Cy7荧光染料量的Free-Cy7和被纳米胶束包埋的SNS-Cy7,在注射后不同时间点,通过活体小动物荧光成像系统,观察在原位白色脂肪的Cy7的荧光量的多少,以判断Cy7在脂肪部位的富集程度。
如图8所示,将脂溶性荧光染料包埋于载辣椒素纳米胶束中后,在脂肪部位原位注射适量的脂溶性荧光染料,其在脂肪部位的富集时间远小于被包埋后的等量的荧光染料的富集持续时间,表明纳米胶束可在脂肪部位缓慢释放包埋的脂溶性物质。
实施例9
首先将3T3-L1细胞接种在12-well细胞培养板上,待细胞长满培养孔后,除了NC组更换正常细胞培养基外,MA组更换正常细胞培养基和MDI药物,诱导细胞分化为白色脂肪细胞(MA),其他组是在形成白色脂肪细胞的基础上给予不同的药物,处理3天后,提取细胞中的全部DNA,然后通过qRT-PCR测定线粒体中mtDNA的拷贝数,可以判断药物对成熟的脂肪细胞中线粒体数量的影响。
如图9所示,在成熟的3T3-L1脂肪细胞(MA)中处理不同药物时,载辣椒素纳米胶束(SNS-Cap组)、不载辣椒素的纳米胶束(SNS组)及白色脂肪棕色化阳性药物罗格列酮(Rosi组)都有效的增加了线粒的拷贝数,说明辣椒素和纳米胶束均具有提高白色脂肪细胞中线粒体的生物发生的效果。
实施例10
为了进一步验证各种药物对成熟脂肪细胞中线粒体呼吸能力的影响,将细胞接种在细胞能量代谢测定专用细胞培养板上,同上,当细胞分化为成熟的白色脂肪细胞后,处理不同的药物后,利用Seahorse仪,首先测定各组的初始细胞呼吸值,测定平行3个点之后加入Oligomycin(一种ATP合酶抑制剂,加入此药后减少的耗氧量代表机体用于ATP合成的耗氧量,间接显示此时细胞的ATP产量),平行测定三个点后,加入FCCP(一种解偶联剂,作为一种质子载体使得大量质子回流,大量耗氧,但是这种质子回流不能形成ATP),FCCP加入后耗氧的增加量代表线粒体的最大耗氧能力,间接显示最大的呼吸能力,而其相对于基础值的高值代表其还具有的呼吸潜力,同样测定3个平行点后加入抗霉素A和寡霉素(R&A),二者是呼吸链抑制剂,完全阻止线粒体耗氧。
如图10所示,在进行细胞能量代谢(Seahorse)的实验结果显示,在3T3-L1成熟白色脂肪细胞中处理载辣椒素纳米胶束及单独的纳米胶束都可以提高线粒体的呼吸活力,提高线粒体的活力。而辣椒素本身或者α-乳白蛋白(α-LAC)本身则不具备该功效。
实施例11
经高脂饲料喂养后,制备肥胖小鼠模型后,脂肪原位注射载辣椒素纳米胶束(IS)或者单独辣椒素(IF)或单独的纳米胶束(SNS),并灌胃载辣椒素纳米胶束(OS)或者单独辣椒素(OF),以一直喂养普通饲料作为阴性对照组(NC),以喂养高脂饲料而不给药的阳性对照组(PC)。
参见图11,一个月之后,发现原位注射的载辣椒素纳米胶束(IS)、灌胃的载辣椒素纳米胶束(OS)以及单独的纳米胶束(SNS)都表现出显著的降低高脂小鼠体重的功效,更加表明纳米胶束本身也具备减脂功效,与前面的体外实验结果相结合,证实纳米胶束通过促进线粒体生物发生及活力,达到与辣椒素协同减肥的功效。
实施例12
将实施例11中各组小鼠的腹股沟白色脂肪取出后提取总蛋白,通过western blot实验验证各实验组中与脂肪分化相关的蛋白(PPARγ,cEBPα)以及辣椒素的直接受体TRPV1的表达。
结果如图12所示,实验结果表明,相对于正常饲料喂养的小鼠(NC),高脂饲料喂养的肥胖模型小鼠的腹股沟白色脂肪细胞中,脂肪分化正相关的PPARγ和cEBPα两种蛋白表达明显上升,表明脂肪合成更活跃,而原位注射SNS-Cap的(IS)组对两种蛋白的抑制作用最明显。而在肥胖小鼠中,TRPV1的表达也有明显下调的趋势,表明辣椒素的接触位点不活跃,而给予原位注射SNS-Cap后,对TRPV1的表达有所恢复,表明IS组可以更多恢复辣椒素的接触位点,帮助辣椒素发挥减脂功效。
在本说明书的描述中,参考术语“一个实施例”、“一些实施例”、“示例”、“具体示例”、或“一些示例”等的描述意指结合该实施例或示例描述的具体特征、结构、材料或者特点包含于本发明的至少一个实施例或示例中。在本说明书中,对上述术语的示意性表述不必须针对的是相同的实施例或示例。而且,描述的具体特征、结构、材料或者特点可以在任一个或多个实施例或示例中以合适的方式结合。此外,在不相互矛盾的情况下,本领域的技术人员可以将本说明书中描述的不同实施例或示例以及不同实施例或示例的特征进行结合和组合。
尽管上面已经示出和描述了本发明的实施例,可以理解的是,上述实施例是示例性的,不能理解为对本发明的限制,本领域的普通技术人员在本发明的范围内可以对上述实施例进行变化、修改、替换和变型。
Claims (2)
1.一种包埋体,其特征在于,包括:
α-乳白蛋白多肽纳米球载体;以及
辣椒素,所述辣椒素包埋于所述α-乳白蛋白多肽载体中;
所述包埋体的粒径大于所述α-乳白蛋白多肽纳米球载体的粒径,其中,所述包埋体和α-乳白蛋白多肽纳米球载体的粒径分别独立地为15~25纳米;
所述包埋体是通过下列方式获得的:
称取α-乳白蛋白6 mg于离心管,加入6 mL 75 mM Tris-HCl缓冲液,所述缓冲液的pH值为7.5,在涡旋振荡器上溶解后,加入地衣芽孢杆菌蛋白酶,所述地衣芽孢杆菌蛋白酶与α-乳白蛋白的质量比为1:70,50°C恒温水浴内,加热30 min,冷却,得到多肽溶液;
在所述多肽溶液中加入20% α-乳白蛋白质量的辣椒素,充分混匀后,将所得混合液用3KD的透析袋透析除去多余的Tris-HCl及辣椒素,将截留物于冻干机冻干后得到所述包埋体。
2.一种抑制成熟白色脂肪细胞中PPAR γ及cEBP α基因的表达的非治疗方法,其特征在于,包括:对成熟白色脂肪细胞施加权利要求1所述包埋体。
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