CN113546097B - 广东虫草子实体乙醇提取物在制备预防肥胖及高血脂药物中的应用 - Google Patents
广东虫草子实体乙醇提取物在制备预防肥胖及高血脂药物中的应用 Download PDFInfo
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Abstract
本发明公开了广东虫草子实体乙醇提取物在制备预防肥胖及高血脂药物中的应用。本发明首次发现广东虫草乙醇提取物能够通过减少高脂饮食肥胖小鼠腹股沟、附睾和肝脏中脂肪含量,缓解其体重的增加;降低其血清中甘油三酯、总胆固醇高密度脂蛋白胆固醇和低密度脂蛋白胆固醇的含量,进而缓解高血脂的症状。在缓解肥胖和降血脂症状方面,广东虫草子实体发挥了积极有效的作用,具有良好的应用前景,极大的扩展了其应用领域。
Description
技术领域:
本发明属于食品和保健品领域,具体涉及广东虫草子实体乙醇提取物在制备预防肥胖及高血脂药物中的应用。
背景技术:
广东虫草Cordyceps guangdongensis(≡Tolypocladium guangdongense)是华南地区特有的新资源食品,与冬虫夏草(Ophiocordyceps sinensis)同属于线虫草科(Ophiocordycipitaceae)。广东虫草子实体安全无毒的,能够进行规模化人工栽培;在腺嘌呤诱导的慢性肾衰竭和慢性支气管炎治疗、抗病毒、抗疲劳等方面具有显著的功效;富含粗蛋白、氨基酸、微量元素及维生素等营养成分。此外,广东虫草子实体含有虫草酸、腺苷、黄酮、皂苷、甾醇等活性成分,具有极大的开发潜力和广阔的应用前景。
2016年,世界卫生组织(WHO)调查结果发现,逾19亿的成年人超重,肥胖人数超过了6.5亿,多于3.4亿名儿童和青少年超重或者肥胖。联合国粮食及农业组织(FAO)2017年调查结果显示,成人肥胖问题比较严重,8个成人中至少有1个人肥胖,肥胖人数超过了6.72亿。肥胖及相关代谢并发症已达到了流行病的程度,世界卫生组织已经宣布肥胖为全球最大的慢性疾病。
肥胖是一个全球日益关注的公共健康问题,是由体内多种代谢紊乱所引起,伴有胰岛素抵抗、高血糖、高血压、血脂异常、动脉硬化等问题,甚至造成二型糖尿病、脂肪肝、肝脏纤维素化等疾病。同时,肥胖也是结肠癌、冠心病、阿尔兹海默病等疾病的潜在影响因素,进而影响人类的寿命。
高血脂是肥胖的主要并发症,是一种全身性疾病,其特征表现为高水平的甘油三酯(TG)、总胆固醇(TC)、高密度脂蛋白胆固醇(HDLC)和低密度脂蛋白胆固醇(LDLC)。
因此,如何有效地预防和治疗肥胖及相关并发症是一个亟需解决的问题,已经引起全世界学者的关注。目前,营养、保健品与医药等方面的研究人员都开展了肥胖和慢性疾病的发生、预防、治疗等相关研究。研究表明冬虫夏草、蛹虫草和蝉花能够降低肥胖小鼠的体重,但对广东虫草子实体缓解肥胖小鼠体重增加、降血脂的研究及相关产品的研发尚未报道。
发明内容:
本发明的目的是提供广东虫草乙醇提取物在制备预防和治疗肥胖及高血脂药物中的应用,为食品科学、保健品、生物医学等领域开发缓解肥胖和降血脂相关产品提供新资源。
广东虫草乙醇提取物能够显著降低高脂饮食诱导的肥胖小鼠的体重,能够减轻棕色脂肪、腹股沟脂肪和附睾脂肪组织的重量;显著降低甘油三酯、总胆固醇、高密度脂蛋白胆固醇和低密度脂蛋白胆固醇的含量;缓解肥胖小鼠肝脏中脂肪的含量及脂滴的积累。
因此本发明提供了广东虫草乙醇提取物在制备预防和治疗肥胖及高血脂药物中的应用。
所述的广东虫草乙醇提取物是将广东虫草子实体在50~60℃烘箱中干燥,并采用超低温粉碎机制成粉,以体积分数50%~60%的食用酒精浸泡提取,取滤液,浓缩干燥获得广东虫草乙醇提取物。
优选,所述的50~60℃烘箱中干燥,其干燥时间为3-4小时。
优选,所述的以体积分数50%~60%的食用酒精浸泡提取,取滤液是以体积分数50%~60%的食用酒精浸泡提取24小时,过滤收集滤液,再用体积分数50%~60%的食用酒精浸泡提取24小时,过滤收集滤液,合并滤液。
优选,所述的浓缩干燥是旋转蒸发仪于52~57℃、55~60rpm条件,减压浓缩干燥。
优选,所述的以体积分数50%~60%的食用酒精浸泡提取是将体积分数50%~60%食用乙醇和广东虫草粉料液按照1:10~15g/mL的比例混合浸泡提取。
本发明的广东虫草乙醇提取物的用法为小鼠灌胃法,用量为每1次/d,每次约为100~150μL,乙醇提取物量为3.4~5.1mg。
本发明首次发现广东虫草乙醇提取物能够通过减少高脂饮食肥胖小鼠腹股沟、附睾和肝脏中脂肪含量,缓解其体重的增加;降低其血清中甘油三酯、总胆固醇高密度脂蛋白胆固醇和低密度脂蛋白胆固醇的含量,进而缓解高血脂的症状。在缓解肥胖和降血脂症状方面,广东虫草子实体发挥了积极有效的作用,具有良好的应用前景,极大的扩展了其应用领域。
附图说明
图1为本发明中广东虫草乙醇提取物对小鼠体重(A)、体重增长率(B)及Less系数(C)的影响。对照组:正常饮食组;高脂组:高脂饮食组;虫草组:高脂饮食+广东虫草乙醇提取物组;*显著性差异(P<0.05);**极显著性差异(P<0.01)。
图2为本发明中广东虫草乙醇提取物对小鼠棕色、腹股沟、附睾脂肪组织重量(A)、体脂比(B)及肝脏重量(C)的影响。对照组:正常饮食组;高脂组:高脂饮食组;虫草组:高脂饮食+广东虫草乙醇提取物组;*显著性差异(P<0.05);**极显著性差异(P<0.01)。
图3为本发明中广东虫草乙醇提取物对小鼠甘油三酯(A)、总胆固醇(B)、高密度脂蛋白胆固醇(C)和低密度脂蛋白胆固醇(D)的影响。对照组:正常饮食组;高脂组:高脂饮食组;虫草组:高脂饮食+广东虫草乙醇提取物组;*显著性差异(P<0.05);**极显著性差异(P<0.01)。图4为本发明中广东虫草乙醇提取物影响小鼠肝脏中脂肪组织的病理切片观察(×200)。对照组:正常饮食组;高脂组:高脂饮食组;虫草组:高脂饮食+广东虫草乙醇提取物组。
具体实施方式
参照附图结合具体实施例对本发明进一步描述。
所述实施例所使用的实验方法若无特殊说明,均为常规实验方法。
所述实施例中所采用的材料、试剂等,若无特殊说明,均可以从商业途径获得。
SPF级雄性C57BL/6J小鼠由深圳市赛百诺基因技术有限公司提供,许可证号:SCXK(京)2019-0008。
广东虫草子实体有广东科华汇泰生物科技有限公司提供。
实施例1
广东虫草乙醇提取物的制备。
广东虫草子实体称取600g,于50~60℃烘箱中干燥3~4h,并采用超低温粉碎机制成广东虫草子实体粉。加入4L体积分数55%食用酒精,浸泡24h,收集滤液,重复一次。合并提取液,采用旋转蒸发仪于55℃、60rpm条件下,减压浓缩干燥获得45.69g固体初提取物。加入1.35L生理盐水,充分混匀,得到广东虫草子实体乙醇提取物溶液,-80℃保存待用。
实施例2
广东虫草乙醇提取物进行干预高脂诱导C57BL/6J肥胖小鼠,具体步骤如下:
1.小鼠分组:90只雄性C57BL/6J小鼠饲养于广东省科学院微生物所实验动物中心,饲养温度为23±1℃,湿度为55±10%,12h光暗交替。适应性培养一周后,将小鼠随机分为9组,每组10只。
2.处理:3组小鼠饲喂常规饲料,6组小鼠饲喂含60%脂肪高脂饲料。饲养13~14周,3组常规饲料组和3组高脂饲料组每个老鼠每天灌胃100μL 0.8%的生理盐水,3组高脂饲料组按照100mg/kg/d的剂量灌胃广东虫草子实体乙醇提取物溶液,每周记录小鼠体重。灌胃24周,直至高脂饲料+生理盐水和常规饲料+生理盐水、高脂饲料+广东虫草子实体乙醇提取物之间小鼠体重出现显著差异,实验期间小鼠自由进食和饮水。
1、广东虫草乙醇提取物对肥胖小鼠体重及Less指数的影响:
将灌胃24周的小鼠禁食24h,称取体重,测量体长(从鼻尖到肛门之间的距离),计算Less指数=(体重)1/3×10/体长。
结果如图1所示,从图1可以看出,广东虫草乙醇提取物干预24周后,与高脂饲料+生理盐水组相比(高脂组),常规饲料+生理盐水组(对照组)和高脂饲料+广东虫草乙醇提取物组(实验组)明显比较轻(图1A)、体重增长率(图1B)及Less指数(图1C)比较小,表明广东虫草乙醇提取物能够缓解高脂饮食诱导的C57BL/6J小鼠体重增加。
2、广东虫草乙醇提取物对肥胖小鼠脂肪组织、肝脏重量的影响:将灌胃24周后的小鼠禁食24h,用乙醚麻醉后,颈椎脱臼处死,取棕色脂肪、腹股沟脂肪、附睾脂肪及肝脏等组织,记录重量并计算体脂比(%)=脂肪质量/小鼠体重×10,具体数据如表1。
表1 广东虫草乙醇提取物对小鼠脂肪、肝脏等组织的影响
注:a,b,c,显著性差异(P<0.05)。
根据表1和附图2发现,广东虫草乙醇提取物可以显著减低肥胖小鼠棕色脂肪的重量,明显缓解高脂饮食诱导肥胖小鼠腹股沟脂肪、附睾脂肪和肝脏的增重,降低肥胖小鼠的体脂率。上述结果表明广东虫草乙醇提取物能够缓解高脂饮食诱导的肥胖小鼠体内脂肪指数的增加。
3、广东虫草乙醇提取物对小鼠血清中甘油三酯、总胆固醇、高密度脂蛋白胆固醇和低密度脂蛋白胆固醇的影响:将灌胃24周后的小鼠禁食24h,用乙醚麻醉并腹腔取血液,室温静置1h,3000rpm离心15min,收集上层血清,根据试剂盒(购于南京建成生物工程研究所)说明书测定血清指标的含量,具体数据如表2。
表2 广东虫草乙醇提取物对小鼠血清指标的影响
注:a,b,c,显著性差异(P<0.05)。
根据表2和附图3可知,广东虫草乙醇提取物缓解了肥胖小鼠中总胆固醇和高密度脂蛋白胆固醇升高的程度,同时显著减低肥胖小鼠血清中甘油三酯和低密度脂蛋白胆固醇的含量,并显著低于常规饲料组该指标的含量。上述结果表明广东虫草乙醇提取物能够影响高脂饮食诱导的肥胖小鼠血清中甘油三酯、低密度脂蛋白胆固醇、总胆固醇和高密度脂蛋白胆固醇含量的变化。
4、广东虫草乙醇提取物对小鼠肝脏中脂肪的影响:将灌胃24周后的小鼠禁食24h,用乙醚麻醉并腹腔取血液,颈椎脱臼处死,取肝脏组织,浸泡在4%多聚甲醛组织固定液中。
将肝脏组织切片,油红染液侵染8~10min(避光);取出切片,停留3s,浸入60%异丙醇分化2次,时间为3s和5s,再依次浸入纯水浸洗2次,各10s;苏木精染色,并用甘油明胶封片剂封片。用电子显微镜(Leica,DM6/DMC4500)观察肝脏中脂肪的变化。
根据附图4,显微镜结果表明,高脂饮食诱导的肥胖小鼠肝脏中积累大量的脂质,呈现明显的橘红色脂滴,且细胞间存在较多膨大的气泡(图4B);而广东虫草乙醇提取物明显缓解肥胖小鼠肝脏中脂质积累,呈现非常淡的橘红色,且细胞间膨大气泡数量显著减少(图4C)。上述结果表明广东虫草乙醇提取物能够有效的缓解肝脏中脂质(如脂滴)的积累。
因此,广东虫草乙醇提取物能够减缓高脂饮食肥胖小鼠体重的增加,减少其血清中甘油三酯、总胆固醇、高密度脂蛋白胆固醇和低密度脂蛋白胆固醇的积累,显著降低其肝脏中脂质(如脂滴)的积累。
综上所述,广东虫草乙醇提取物在缓解肥胖、降血脂及改善相关慢性疾病方面发挥了积极的作用。本发明提供了广东虫草乙醇提取物的制备方法以及减重、减脂和改善代谢紊乱方面的用途,为生物医药、食品、保健品等领域产品开发提供一种新资源。
以上列举的仅为本发明的若干实施例。然而,本发明不限于以上实施例,还有许多变形。该领域的普通技术人员能从本发明公开的内容直接导出或联想到的所有变形,均应认为是本发明的保护范围。
Claims (1)
1.广东虫草乙醇提取物在制备预防和治疗肥胖及高血脂药物中的应用;所述的广东虫草乙醇提取物是将广东虫草子实体称取600 g,于50~60℃烘箱中干燥3~4 h,并采用超低温粉碎机制成广东虫草子实体粉,加入4 L体积分数55%食用酒精,浸泡24 h,收集滤液,重复一次,合并提取液,采用旋转蒸发仪于55℃、60 rpm条件下,减压浓缩干燥获得固体初提取物,加入1.35 L生理盐水,充分混匀,得到广东虫草子实体乙醇提取物溶液。
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