CN113041263B - 一种贻贝功能性脂质在制备治疗2型糖尿病产品中的应用 - Google Patents
一种贻贝功能性脂质在制备治疗2型糖尿病产品中的应用 Download PDFInfo
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Abstract
本发明属于医药领域,涉及一种贻贝功能性脂质在治疗2型糖尿病产品中的应用,所述贻贝功能性脂质为从贻贝中提取的含有多种不饱和脂肪酸的功能性活性物质;通过双盲随机对照实验,研究贻贝功能性脂质的摄入对2型糖尿病的影响,招募2型糖尿病患者,随机分为3组,分别采用贻贝功能性脂质、鱼油和玉米油进行干预,共干预60天;在基线和第60天抽取空腹血样。申请人首次发现,贻贝功能性脂质补充对中国2型糖尿病患者的血糖控制有潜在的改善作用,为应用贻贝功能性脂质控制中国2型糖尿病患者的血糖提供了新的证据,也将为贻贝功能性脂质类功能性食品的开发提供理论及技术基础,对于促进我国功能性食品产业的发展具有重要意义。
Description
技术领域:
本发明属于医药领域,涉及一种贻贝功能性脂质在制备治疗2型糖尿病产品中的应用,通过双盲随机对照实验,研究贻贝功能性脂质的摄入对2型糖尿病的影响,结果表明贻贝功能性脂质对治疗2型糖尿病效果显著。
背景技术:
贻贝属于软体动物门贻贝科,是我国传统的养殖贝类,以生长快、繁殖力强、适应力强,易于人工养殖等特点,被广泛养殖,其中经济价值高的有翡翠贻贝、紫贻贝、厚壳贻贝等品种。我国是全球最大的贻贝产地,年产超过80万吨。我国贻贝海水养殖主要集中在山东、浙江、福建等地,2019年山东省贻贝海水养殖产量最高,达到38.41万吨。然而,贻贝在我国却没有得到充分的利用,被认为是一种低产值海产品。目前,贻贝类主要是冷冻品和干制、罐制、调味品等传统产品,产业起步较晚、缺乏品牌、产业链短小。贻贝也是在开采后以批发零售、市场鲜销、简单初加工为主,产品以全壳、半壳、贻贝肉等为主的单一加工为多,在精深加工和综合利用还是缺乏实质性进展,所以需要在组建技术团队、养殖高新技术、产品精深加工、冷链物流、贝壳污染等各方面不断进行贻贝类加工研发,高效地利用贻贝类。
此外,贻贝也具有重要的药用价值。例如,在中国申请的专利CN201810666201.3公开的一种贻贝提取物的制备工艺及其应用,主要经过脱臭除脂、清洗、变性、酶解、浓缩和干燥等操作对贻贝壳这一副产物进行了利用;所得贻贝提取物具有无菌、高活性和易储存等特点;通过大鼠进行喂养试验,通过核磁共振扫描检测各组大鼠左后足骨关节肿胀情况,结果显示喂食贻贝肽大鼠骨关节腔积液和肿胀都有所缓解;在完全弗氏佐剂诱导的急性RA中,贻贝肽可以通过其抗炎和消水肿作用缓解RA的症状,防止病情进一步恶化;在胶原诱导的慢性RA中,贻贝肽可通过其抗炎和消水肿作用达到一定的预防效果;在长期喂食贻贝肽的过程中并没有观察到大鼠出现传统治疗RA药物所引起的不良反应;中国专利申请CN201810910720.X公开了一种贻贝类胡萝卜素提取物在保护眼睛健康中的应用,其中类胡萝卜素提取物的制备步骤如下:将贻贝干燥并压碎后,酶解,然后将酶解液过滤,取滤液离心,再将过滤所得贻贝残渣和离心沉淀混合,加入有机提取剂浸提,最后将浸提液抽滤所得滤液浓缩,即得类胡萝卜素提取物;类胡萝卜素提取物可制成还含有外加油相或固相分散载体的组合物;所制提取物及其组合物可在保护眼睛健康中应用;还可用于制备抗氧化物、增强免疫功能、防止皮肤衰老、保护中枢神经、预防心脑血管疾病和缓解运动疲劳等领域的应用。
慢性炎症在糖尿病的发生发展过程中发挥着重要作用。由于贻贝中含有抗炎活性成分,所以有必要对贻贝提取物对糖尿病患者的影响进行研究,但目前还未有关于贻贝功能性脂质对2型糖尿病有治疗作用的报道。
发明内容:
本发明为了克服现有技术存在的缺点,提供一种贻贝功能性脂质在制备治疗2型糖尿病产品中的应用,通过随机对照实验,研究表明贻贝功能性脂质的摄入能够降低中国2型糖尿病患者的血糖和炎症反应。
为了实现上述目的,本发明提供一种贻贝功能性脂质在制备治疗2型糖尿病产品中的应用,所述贻贝功能性脂质为从贻贝中提取的含有多种不饱和脂肪酸的功能性活性物质。
所述产品包括药物、保健品或食品。
本发明与现有技术相比,发明人首次研究发现贻贝功能性脂质能够降低2型糖尿病患者的血脂和炎症,改善2型糖尿病患者的血糖控制;通过人体干预实验直接评价贻贝功能性脂质的摄入对2型糖尿病的影响以提供临床上的直接证据,对于2型糖尿病的营养治疗,对于降低我国2型糖尿病的发病率和改善我国居民健康状况具有重要意义。同时,本发明也将为贻贝功能性脂质类功能性食品的开发提供理论及技术基础,对于促进我国功能性食品产业的发展具有重要意义。
说明书附图:
图1为本发明实施例2涉及的贻贝功能性脂质的摄入对2型糖尿病的影响人体随机对照干预试验流程示意图。
具体实施方式:
下面通过具体的实施例并结合附图对本发明作进一步说明。
实施例1:
本实施例涉及一种贻贝功能性脂质的制备方法,具体步骤为:
1、贻贝预处理:将紫贻贝清洗、去壳、留肉;然后在贻贝肉中加入酒石酸作为稳定剂,充分搅拌均匀,接着进行搅碎处理,得厚壳贻贝碎肉,整个贻贝预处理在0~15℃的温度下进行,酒石酸与贻贝的重量比为1%~7%;
2、冷冻干燥:先将贻贝碎肉进行冷冻干燥,将贻贝肉平铺至厚度为3-15mm,冻结时间4-6h,冻结温度-20~-30%,冷阱温度-40~-70%℃,真空度10~50pa,冻干最终温度为20-30℃,干燥时间15~24h,然后将所得的冻干贻贝肉在粉碎机中粉碎,得到贻贝肉冻干粉;
3、超临界CO2萃取:将贻贝肉冻干粉进行超临界CO2萃取,分离釜Ⅰ的压力为7~8MPa,纬度为60℃,分离釜Ⅱ的压力为4~6MPa,温度为35℃,萃取釜压力为15~35MPa,温度为35~55℃,CO2流量为萃取釜容量的10~20倍/h,萃取时间为1~3h,每隔15~45min收集脂溶性提取物,得到贻贝脂溶性提取物,即贻贝功能性脂质,贻贝功能性脂质为含有多种不饱和脂肪酸的功能性活性物质。
实施例2:
本实施例涉及贻贝功能性脂质在制备治疗2型糖尿病产品中的应用试验,具体为贻贝功能性脂质的摄入对2型糖尿病的影响的人体随机对照干预试验。
1、实验材料
(1)本实验已经在中国临床试验注册中心完成注册(ChiCTR1900025617)。试验获得了青岛大学附属医院伦理委员会的批注(QYFYWZLL25550)。所有参与者都完成了书面知情同意书。
(2)试验胶囊制备:
将上海展望生物科技有限公司提供的鱼油和玉米油与实施例1制备的贻贝功能性脂质(简称贻贝油组)胶囊每克标准化,外观相同,每粒胶囊0.8克。每粒鱼油胶囊含有30%的EPA+DHA,每粒贻贝油胶囊同样含有30%的EPA+DHA。所有胶囊均装在不通光的棕色瓶中(60粒胶囊/瓶),三种胶囊分别标记为A、B、C油。在干预过程中,所有参与者或护士/医生都不知道油的类型。
(3)研究参与者的纳入和排除标准:
纳入标准为:1)空腹血糖≥7.0mmol/L。空腹的定义为至少八小时内无热量摄入;2)糖化血红蛋白HbAlc≥7.0%(≥53mmol/mol);3)口服糖耐量测试时2小时血糖≥11.1mmol/L;4)年龄至少40岁。
排除标准为:1)维生素和矿物质吸收异常(血清维生素或矿物质水平异常,或有临床病史);2)在过去3个月内使用维生素和矿物质补充剂;3)在过去一个月内服用鱼油、鱼肝油等补充剂;4)严重的肾脏、肝脏、心脏或精神疾病(糖尿病并发症除外);5)每日注射两针及以上胰岛素;6)有癌症、甲状腺疾病或酗酒史,以及怀孕或哺乳史;7)在筛检前30天内参加其他临床试验。
2、实验过程
(1)参与者的随机化和干预方法:
将所有纳入的研究对象共133名均随机分为鱼油组(n=44)、玉米油组(n=45)和贻贝油组(n=44),采用STATA软件根据年龄、性别随机分配序列生成。每一组的受试者均要求每天服用2粒胶囊,分别在实验开始前和实验结束时抽取空腹血样。干预期间,所有患者被要求保持他们的日常饮食,生活方式和使用处方药物,并避免使用n-3脂肪酸补充剂。在60天的干预治疗后,患者回到研究中心进行最后的现场检查。
(2)实验周期:
本实验计划连续干预时间为90天,试验开始后30天中断,并于3个月后重新开始,第二阶段共连续干预60天,且在第二阶段开始前抽取所有参与者的空腹血样(10mL),所以本次实验的最终实验周期为60天。
3、实验分析
(1)生物化学参数的测定:
在基线(第0天)和第60天抽取了所有参与者的空腹血样(10mL)。血清高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、总胆固醇、甘油三酯、葡萄糖、胰岛素在日立7020化学分析仪上分析。血清胰岛素测定采用美国伊利诺伊州雅培公园Abbott实验室的Architect胰岛素试剂盒。以空腹血糖和胰岛素为指标,用空腹血糖(mmol/L)×胰岛素(mU/L)/22.5公式计算胰岛素抵抗的稳态模型评价(HOMA-IR)。HOMA-IR模型被用来表示胰岛素抵抗,而不是HOMA2模型,因为它在中国人群中的应用更广泛,因此在中国的相关研究中也更具可比性。采用全自动血红蛋白A1c分析仪测定糖化血红蛋白A1c(HbA1c)。采用酶联免疫吸附法试剂盒测定白细胞介素10(IL-10)、丙二醛(MDA)、白介素-1β(IL-1β)和肿瘤坏死因子-a(TNF-a)。未立即测量的血样保存在-80℃以下以作进一步分析。在基线时测量体重、身高、腰围和臀围等人体测量参数。
(2)统计学分析:
在SPSS(Version 22)软件中进行了所有的统计分析。总样本量计算是基于80%权重(α双侧=0.05)检测不同HOMA-IR 20%或0.63之间(SD=1.1)组(n=150,50个/组),考虑20%的失访率(n=187,62/组),基于我们之前的工作。这个样本大小(n=50/组)也将使我们能够有84%的权重(α双侧=0.05)检测不同组间糖化血红蛋白20%或12mmol/mol(SD=20)的差异。
检查所有结果变量是否为正态分布,若为非正态分布则进行log变换(HOMA-IR、葡萄糖、胰岛素、糖化血红蛋白、TG为自然对数变换)。采用单因素方差分析(连续变量)或卡方检验(分类变量)检验各组基线差异与变化量的差异。采用t检验分析基线与第90天之间的差异。
4、实验结果
实验开始前(在基线时)的所有参与者的人体测量数据和血液检测数据见表1,没有观察到三个干预组参与者之间的血脂或血糖特征有任何显著差异。在干预期间,鱼油组的3名参与者、贻贝油组的1名参与者和玉米油组的1名参与者退出了试验,鱼油组、贻贝油组和玉米油组的全程参与者分别为41人、43人和44人。实验结果以全程参与者的数据为有效数据进行分析。
表1研究参与者的基线特征
项目 | 单位 | 鱼油组(n=44) | 贻贝油组(n=44) | 玉米油组(n=45) | P值a |
年龄 | Age,y | 59.94±9.98 | 62.75±9.22 | 61.94±9.52 | 0.321 |
女性占比 | Women,n(%) | 29(49) | 30(52) | 33(49) | 0.607 |
体重 | Weight,kg | 69.14±12.02 | 69.67±10.82 | 68.38±10.43 | 0.843 |
体质质量指数 | BMI,kg/m2 | 25.95±3.26 | 26.12±3.12 | 25.95±3.43 | 0.955 |
血红蛋白 | HGB,g/L | 141.41±13.87 | 140.71±12.03 | 140.36±13.55 | 0.929 |
糖化血红蛋白 | HBA1C,% | 8.22±2.13 | 7.58±1.70 | 7.74±1.63 | 0.235 |
血糖 | GLU,mmol/L | 8.41±3.00 | 7.24±2.57 | 7.88±2.99 | 0.161 |
胰岛素 | INS,μU/mL | 15.08±30.77 | 12.47±10.78 | 11.03±7.19 | 0.604 |
甘油三酯 | TG,mmol/L | 2.23±1.51 | 2.51±2.05 | 2.25±1.88 | 0.733 |
胆固醇 | CHOL,mmol/L | 5.66±1.25 | 5.42±1.16 | 5.70±1.02 | 0.454 |
高密度脂蛋白胆固醇 | HDL-C,mmol/L | 1.56±0.31 | 1.47±0.31 | 1.53±0.30 | 0.353 |
低密度脂蛋白胆固醇 | LDL-C,mmol/L | 2.73±0.74 | 2.64±0.70 | 2.83±0.68 | 0.454 |
肿瘤坏死因子 | TNF-a,nmol/L | 12.77±3.2 | 12.68±2.96 | 11.78±2.89 | 0.234 |
白细胞介素1β | IL-1β,nmol/L | 11.03±2.65 | 11.36±2.92 | 11.39±2.89 | 0.804 |
白细胞介素10 | IL-10,nmol/L | 142.03±45.69 | 140.34±39.82 | 147.48±46.86 | 0.729 |
丙二醛 | MDA,nmol/ | 1.29±0.28 | 1.10±0.32 | 1.07±0.24 | 0.01 |
数据以均数±标准差表示;a组间差异。
4.1贻贝功能性脂质对2型糖尿病患者血糖及血脂的影响
贻贝功能性脂质对2型糖尿病患者血糖及血脂结果见表2,从表2可以看出三组间的甘油三酯变化量、高密度脂蛋白存在显著性差异(P2值分别为0.005、0.015),与玉米油组和鱼油组相比,贻贝油组的甘油三酯水平显著降低(P1=0.007),贻贝油组的高密度脂蛋白降低0.46mmol/L,而玉米油组和鱼油组分别降低0.45nmol/L和0.38mmol/L。干预60天后,贻贝油组的胰岛素、胰岛素抵抗稳态模型(HOMA-IR)、甘油三酯、胆固醇显著降低(P1值分别为0.005、0.026、0.007和<0.001),而其他两组未见显著性差异。
表2贻贝功能性脂质对2型糖尿病患者血脂、血糖等指标的影响
数据以均数±标准差表示;1组内差异;2组间差异。
4.2贻贝功能性脂质对2型糖尿病患者炎性因子的影响
贻贝功能性脂质对2型糖尿病患者炎性因子的影响分析结果见表3,从表3可以看出,三组间的白细胞介素1β存在显著性差异(P2=0.002),贻贝油组的白细胞介素1β降低2.17mmol/L,而玉米组和鱼油组的白细胞介素1β分别降低1.9nmol/L和升高0.04mmol/L。干预60天后,贻贝油组的肿瘤坏死因子α和白细胞介素1β显著降低(P1值分别为0.01以及0.003),而其他两组未见显著性差异。
表3贻贝功能性脂质对2型糖尿病患者炎性因子指标的影响
数据以均数±标准差表示;1组内差异;2组间差异。
综上,本实施例的随机对照试验表明,60天的贻贝功能性脂质补充对中国2型糖尿病患者的血糖控制有潜在的改善作用。贻贝功能性脂质补充剂能够在降低胰岛素方面改善2型糖尿病患者的血糖控制,胰岛素是机体内唯一降低血糖的激素,同时促进糖原、脂肪、蛋白质合成。此外,贻贝功能性脂质持续降低这些患者的血脂和炎症,包括甘油三酯、胆固醇、高密度脂蛋白、肿瘤坏死因子α和白细胞介素1β。在服用鱼油和玉米油补充剂的患者中,没有发现对血糖特征有显著影响。本发明为应用贻贝功能性脂质控制中国2型糖尿病患者的血糖提供了新的证据。
Claims (3)
1.一种贻贝功能性脂质在制备治疗2型糖尿病产品中的应用,其特征在于,所述贻贝功能性脂质的制备方法为:贻贝预处理,将贻贝清洗、去壳、留肉;将贻贝碎肉进行冷冻干燥,然后将所得的冻干贻贝肉在粉碎机中粉碎,得到贻贝肉冻干粉;贻贝肉冻干粉进行超临界CO2萃取,得到贻贝脂溶性提取物,即为贻贝功能性脂质;所述产品为药物或药物组合物。
2.根据权利要求1所述的贻贝功能性脂质在制备治疗2型糖尿病产品中的应用,其特征在于,所述贻贝功能性脂质为从贻贝中提取的含有多种不饱和脂肪酸的功能性活性物质。
3.根据权利要求1所述的贻贝功能性脂质在制备治疗2型糖尿病产品中的应用,其特征在于,通过以下应用步骤验证获得:
通过双盲随机对照实验,研究贻贝功能性脂质的摄入对2型糖尿病的影响:招募2型糖尿病患者,随机分为3组,分别采用贻贝中提取的贻贝功能性脂质、鱼油以及玉米油进行干预,每人每天1.6g,干预周期60天,分别于第0天、第60天取2型糖尿病患者空腹血,分离血清。
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