CN112316013A - 一种含番石榴和宝乐果的组合物及其应用 - Google Patents
一种含番石榴和宝乐果的组合物及其应用 Download PDFInfo
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- CN112316013A CN112316013A CN202011130170.3A CN202011130170A CN112316013A CN 112316013 A CN112316013 A CN 112316013A CN 202011130170 A CN202011130170 A CN 202011130170A CN 112316013 A CN112316013 A CN 112316013A
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Abstract
本发明公开了一种含番石榴和宝乐果的组合物及其应用，该组合物含番石榴叶提取物8‑12份、番石榴果粉16‑32份、宝乐果果粉16‑32份、宝乐果叶提取物8‑12份、黄芪提取物12‑18份和Vc 8‑12份；所述的宝乐果叶提取物，由以下步骤制得：取宝乐果植物的干叶，粉碎，用稀乙醇渗滤提取，滤出提取液，经大孔树脂富集，90％乙醇洗脱，洗脱液浓缩、喷雾干燥制粉，得到宝乐果叶提取物。本发明组合物活性物质明确、作用机制清楚；首次将宝乐果和宝乐果提取物，与番石榴、黄芪、Vc组方，多成分协同增效、多靶点作用降血糖、降血脂和防治糖尿病肾病。
Description
技术领域
本发明属于植物天然药物领域，具体涉及一种含番石榴和宝乐果的组合物及其应用。
背景技术
糖尿病(diabetes mellitus，DM)是一种严重危害人类健康的内分泌代谢疾病，已经有大量的研究表明胰岛素抵抗是2型糖尿病最重要的发病机制之一。此外，糖耐量低减、中心性肥胖、高血压、高甘油三酯血症、动脉粥样硬化、胆固醇浓度下降、低密度脂蛋白质量下降(颗粒致密变小)、肥胖基因编码蛋白瘦素的增高、低体重儿以及生长激素缺乏等都伴有胰岛素抵抗。胰岛素抵抗以及由此引起的代偿性高胰岛素血症与人群中2型糖尿病、高脂血症、高血压、冠心病、糖尿病肾病(diabetic nephropathy，DN)的高发生率密切相关，糖尿病并发症已成为继心脑血管疾病和肿瘤之后人类第三大致死原因。治疗胰岛素抵抗已经成为预防、控制糖尿病及其并发症的关键。
番石榴(Psidium guajava L)为桃金娘科番石榴属植物，广泛分布于我国岭南地区。宝乐果为茜草科林果属植物Borojoa patinoi Cuatrec和Borojoa sorbilis Cuatrec的成熟水果，生长在南美洲厄瓜多尔和哥伦比亚，富含天然环烯醚萜苷类、多酚和黄酮类，以及B族维生素等。黄芪(Astragalus membranaceus(Fisch.)Bge.)为豆科植物，含有黄芪多糖、黄芪皂苷、黄芪黄酮等多种活性成分。
发明内容
本发明的目的在于提供一种含番石榴和宝乐果的组合物，及其在药物、保健品和食品中的应用。
本发明的目的通过下述技术方案实现：
一种组合物，包括以下重量份数的成分：
优选地，所述的组合物包括以下重量份数的成分：
所述的番石榴叶提取物(FSL)，由以下步骤制得：
将干燥的番石榴叶粉碎，用稀乙醇渗滤提取，滤出提取液，经大孔树脂富集，90％乙醇洗脱，洗脱液浓缩、喷雾干燥制粉，得到番石榴叶提取物，含总三萜酸；
所述的宝乐果叶提取物(BLG)，由以下步骤制得：
取宝乐果植物的干叶，粉碎，用稀乙醇渗滤提取，滤出提取液，经大孔树脂富集，90％乙醇洗脱，洗脱液浓缩、喷雾干燥制粉，得到宝乐果叶提取物(BLG)，含有总环烯醚萜苷；
所述的稀乙醇，是体积分数40-45％的乙醇溶液；
所述的大孔树脂优选D101大孔树脂。
所述的黄芪提取物(HQZ)，由以下步骤制得：
取干燥的黄芪根粉碎，水渗滤提取，滤出提取液，浓缩、喷雾干燥制粉，得含有多糖、皂苷、黄酮的黄芪提取物；
所述的渗滤提取，渗漉速度为1～3ml/min。
本发明的番石榴叶提取物(FSL)中，总三萜酸(FSL)为番石榴叶降血糖的活性物质，FSL能够显著降低糖尿病大鼠的血糖水平，提高糖尿病大鼠脂肪组织过氧化物酶体增生物激活受体(PPARγ)蛋白的表达水平，同时下调PTP1B蛋白的表达，促进胰岛素的信号传导。FSL能够显著降低糖尿病大鼠的血脂水平，明显改善肾功能损伤。
本发明的宝乐果叶提取物(BLG)中，总环烯醚萜苷(BLG)为宝乐果叶降血糖的活性物质，BLG能够显著降低糖尿病大鼠的血糖水平，上调脂肪细胞过氧化物酶体增生物激活受体(PPARγ)的表达水平，显著增加培养液中葡萄糖的消耗。BLG能够显著降低糖尿病大鼠的血清总胆固醇和血清中甘油三酯水平，具有减轻糖尿病肾病变的作用。
本发明所述的黄芪提取物(HQZ)，可升高模型小鼠的白细胞和红细胞数目，增强小鼠迟发性变态反应，增强机体的免疫和细胞免疫功能。
胰岛素抵抗是2型糖尿病最重要的发病机制之一，是指组织对胰岛素的敏感性下降,代偿性引起胰岛β细胞分泌胰岛素增加，其实质为胰岛素介导的细胞糖代谢能力的减低。发生胰岛素抵抗的主要部位是依赖胰岛素的葡萄糖利用器官，如骨骼肌、肝脏、脂肪组织，当没有足够的胰岛素以增强胰岛素敏感性时，葡萄糖稳态遭到破坏,就会出现葡萄糖耐量减低，以致2型糖尿病的发生，并导致多个器官水平的缺陷。维护肝、肾、神经细胞、毛细血管的功能，以及增强免疫功能，一是防止糖尿病并发症的出现；另，这些器官和功能的完好对避免或延缓糖尿病的发生发展又具有重要的现实意义。
本发明的组合物可以用于制备药物、保健品和食品；
优选地，所述的药物具有降血糖、提高胰岛素水平、消除或减轻胰岛素抵抗(提高胰岛素敏感指数)、降低糖化血清蛋白水平、降低血脂含量或保护肾脏中的一种以上功效；
优选地，所述的药物是治疗2型糖尿病的药物；
所述的药物还可以含有一种以上药学上可以接受的载体；所述的载体优选为缓释剂、赋形剂、填充剂、粘合剂、湿润剂、崩解剂、吸收促进剂、吸附载体、表面活性剂或润滑剂等；
所述的药物可以进一步制成片剂、颗粒剂或胶囊等多种形式，各种剂型的药物可以按照药学领域的常规方法制备。
本发明相对于现有技术具有如下的优点及效果：
本发明中，三种植物提取物的制备工艺简单稳定、成本低、易产业化。本发明组合物活性物质明确、作用机制清楚；首次将宝乐果和宝乐果提取物，与番石榴、黄芪、Vc组方，多成分协同增效、多靶点作用降血糖、降血脂和防治糖尿病肾病。
附图说明
图1是糖尿病大鼠肾脏切片结果。
图2是糖尿病大鼠脂肪组织过氧化物酶体增殖物激活受体(PPARγ)蛋白的表达量；其中，Control为正常对照组，Model为模型组，BF-L为组合物BF低剂量组，BF-M为组合物BF中剂量组，BF-H为组合物BF高剂量组，MF为二甲双胍组。
具体实施方式
下面结合实施例及附图对本发明作进一步详细的描述，但本发明的实施方式不限于此。
实施例1
含番石榴和宝乐果组合物(BF)的颗粒剂的制备，包括以下步骤：
(1)干燥的番石榴叶和宝乐果叶各10.0kg，粉碎后，分别用40％乙醇100L渗滤提取，滤出提取液，经D101大孔树脂富集，90％乙醇洗脱，洗脱液浓缩、喷雾干燥制粉，分到得到番石榴叶提取物和宝乐果叶提取物；
(2)干燥的黄芪根12.0kg，粉碎，120L水渗滤提取，滤出提取液，喷雾干燥制粉，得到黄芪提取物；
(3)新鲜的番石榴和宝乐果果实去除种子，果肉各7.5kg，分别匀浆，过筛，喷雾干燥制粉，分别得到番石榴果粉和宝乐果果粉；
(4)取番石榴叶提取物(FSL)500g，番石榴果粉1000g，宝乐果果粉1000g，宝乐果叶提取物500g，黄芪提取物750g，Vc 500g，糊精750g，粉碎混合均匀，灭菌，按照每包1.0±0.02g的剂量包装。
实施例2
含番石榴和宝乐果组合物(BF)的的咀嚼片的制备，包括以下步骤：
(1)干燥的番石榴叶和宝乐果叶各20.0kg，粉碎后，用45％乙醇200L渗滤提取，滤出提取液，经D101大孔树脂富集，90％乙醇洗脱，洗脱液浓缩、喷雾干燥制粉，分到得到番石榴叶提取物和宝乐果叶提取物；
(2)干燥的黄芪根25.0kg，粉碎，200L水渗滤提取，滤出提取液，喷雾干燥制粉，得到黄芪提取物；
(3)新鲜的番石榴和宝乐果果实去除种子，果肉各15.0kg，分别匀浆，过筛，喷雾干燥制粉，分别得到番石榴果粉和宝乐果果粉；
(4)取番石榴叶提取物1000g，番石榴果粉2000g，宝乐果果粉2000g，宝乐果叶提取物1000g，黄芪提取物1500g，Vc 1000g，微粉硅胶400g、羟甲基纤维素钠1600g混合均匀，用10％预胶化淀粉浆作为粘合剂，湿法造粒，烘干，加入适量的硬脂酸镁混合，压制成40000片片剂。
实施例3、组合物BF对2型糖尿病大鼠的作用
试验方法：
将90只雄性SD大鼠(4周龄清洁级，体重155～180g，由广东省动物中心提供，随机分为正常对照组10只和糖尿病模型组80只，正常组给予基础饲料，模型组给予高脂饲料(猪油20％，蔗糖20％，蛋黄3％，基础饲料57％)。
喂养4周后，禁食12h后，用0.1mmol/L枸橼酸钠缓冲液(pH值4.2)将STZ(链脲佐菌素)配成2％的溶液，按35mg·kg-1腹腔注射，一周后测大鼠2h葡萄糖耐受量(禁食12h，灌胃给予2g/kg的葡萄糖，测灌胃后2h的血糖)，筛选血糖值大于11.1mmol/L的大鼠确定为DM大鼠。
稳定一周后将成模的DM大鼠70随机分为7组，即组合物高剂量组(400mg/kg)、中剂量组(200mg/kg)、低剂量组(100mg/kg)、二甲双胍Metformin组(300mg/kg)、模型组(生理盐水2ml/kg)，每组各10只。每天灌胃给药1次，连续6周。每周末称量体重1次，根据体重调整给药剂量。于实验结束前1天，大鼠禁食12h，取血测各指标。
试验结果：
表1.组合物BF对糖尿病大鼠空腹血糖(FBG)、空腹胰岛素(FIN)，胰岛素敏感指数(ISI)的影响(X±s，n＝8)
#p<0.05，##p<0.01，模型组νs正常对照组；*p<0.05，**p<0.01，给药组νs模型组。
ISI为空腹血糖与空腹胰岛素乘积的倒数，分析时取自然对数。
与正常对照组比较，模型组空腹血糖水平显著升高(p<0.01)，空腹胰岛素和胰岛素敏感指数显著降低(p<0.05，或p<0.01)。与模型组相比，组合物BF随着剂量增加，可显著降低糖尿病大鼠空腹血糖水平，提高胰岛素水平和胰岛素敏感指数(p<0.05，或p<0.01)。
表2.组合物对糖尿病大鼠糖化血清蛋白(GSP)的影响(X±s，n＝8)
##p<0.01，模型组νs正常对照组；**p<0.01，给药组νs模型组。
糖化血清蛋白(GSP)，反映测量前1-3周的血糖水平。与正常对照组比较，模型组GSP显著升高(p<0.01)；二甲双胍组和组合物BF各剂量组的GSP与模型组相比均明显下降(p<0.01)。
表3.组合物对糖尿病大鼠甘油三脂(TG)、总胆固醇(TC)、游离脂肪酸(FFA)的影响(X±s,n＝8)
#p<0.05，##p<0.01，模型组νs正常对照组；*p<0.05，**p<0.01，给药组νs模型组。
与正常对照组比较，模型组TG、TC及FFA显著升高(p<0.01)；二甲双胍组、组合物高、中、低剂量组与模型组比较，其TG、TC及FFA均显著降低，具有统计学意义(p<0.05，或p<0.01)。
表4.组合物对糖尿病大鼠肌酐(CR)、尿素氮(BUN)的影响(X±s，n＝8)
##p<0.01，模型组νs正常对照组；*p<0.05，**p<0.01，给药组νs模型组。
与正常对照组比较，模型组和二甲双胍组血清CR、BUN显著升高(p<0.05，或p<0.01)。组合物与模型组比较，其CR有降低的趋势，但无统计学意义；组合物高剂量组可显著降低BUN(p<0.05)。提示组合物对肾功能有一定的保护作用。
同时，糖尿病大鼠肾脏切片结果见图1。正常组大鼠肾小球形态完整，轮廓清晰，肾小管未见异常。模型组大鼠肾小球形态不规则，肾小囊变窄，系膜细胞增生，系膜区增宽，肾小管空泡变性增大，胞质空亮，可见散在的蛋白颗粒。给药组大鼠的病理损害有所改善，肾小球轮廓较规则，系膜细胞增生好转，肾小管空泡变性情况有所减少。
组合物BF对糖尿病大鼠脂肪组织过氧化物酶体增殖物激活受体(PPARγ)蛋白表达的影响见图2。激动PPARγ蛋白，可增强胰岛素在外周组织的敏感性，减轻胰岛素抵抗，从而达到降血糖的作用。与正常对照组比较，糖尿病模型组大鼠PPARγ蛋白表达显著下降(p≤0.01)。给药后，组合物BF或二甲双胍可显著升高糖尿病大鼠脂肪组织的PPARγ蛋白表达，与模型组相比，具有统计学显著性意义(p≤0.05)。
实施例4、组合物BF对3T3-L1脂肪细胞葡萄糖和脂肪酸代谢的影响试验方法：
将3T3-Ll前脂肪细胞(购自中国医学科学院细胞中心)转入96孔板中，调整细胞密度为5×106mL-1，诱导分化后，分为6组，即溶媒对照组(1‰DMSO)、组合物BF组(3、10、30、100μg/L)、二甲双胍组(10μM)，每组6个复孔，实验重复三次。培养12h后，更换培养基并加药物进行干预，药物作用于96孔板中的细胞24小时，葡萄糖试剂盒GOD-POD法(葡萄糖氧化酶法)检测上清液中葡萄糖的浓度，用对照组的葡萄糖浓度减去不同给药组葡萄糖的浓度计算出葡萄糖的消耗量。酶联免疫吸附法测培养上清液中游离脂肪酸(FFA)的浓度。
试验结果：
表5.组合物对3T3-L1脂肪细胞葡萄糖消耗、游离脂肪酸产生的影响(X±s，n＝6)
与溶媒对照组比较*P<0.05，**P<0.01；与不含胰岛素组比较#P<0.05，##P<0.01。
与溶媒对照组相比，组合物BF和二甲双胍均能显著增加3T3-L1脂肪细胞24h葡萄糖消耗，减少脂肪细胞游离脂肪酸(FFA)的产生。在3-100μg/L剂量范围，组合物随着浓度增加，葡萄糖消耗量逐渐增加，呈一定的剂量依赖性。组合物在3-100μg/L剂量时均显著增加胰岛素刺激下脂肪细胞葡萄糖的消耗量。
上述实施例为本发明较佳的实施方式，但本发明的实施方式并不受上述实施例的限制，其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化，均应为等效的置换方式，都包含在本发明的保护范围之内。
Claims (10)
1.一种组合物，其特征在于包括以下重量份数的成分：
所述的番石榴叶提取物，由以下步骤制得：
将干燥的番石榴叶粉碎，用稀乙醇渗滤提取，滤出提取液，经大孔树脂富集，90％乙醇洗脱，洗脱液浓缩、喷雾干燥制粉，得到番石榴叶提取物；
所述的宝乐果叶提取物，由以下步骤制得：
取宝乐果植物的干叶，粉碎，用稀乙醇渗滤提取，滤出提取液，经大孔树脂富集，90％乙醇洗脱，洗脱液浓缩、喷雾干燥制粉，得到宝乐果叶提取物。
2.根据权利要求1所述的组合物，其特征在于包括以下重量份数的成分：
3.根据权利要求1所述的组合物，其特征在于：所述的黄芪提取物，由以下步骤制得：
取干燥的黄芪根粉碎，水渗滤提取，滤出提取液，浓缩、喷雾干燥制粉，得到黄芪提取物。
4.根据权利要求1所述的组合物，其特征在于：所述的稀乙醇，是体积分数40-45％的乙醇溶液。
5.权利要求1-4任一项所述的组合物在制备药物、保健品和食品中的应用。
6.根据权利要求5所述的应用，其特征在于：所述的药物具有降血糖、提高胰岛素水平、消除或减轻胰岛素抵抗、降低糖化血清蛋白水平、降低血脂含量或保护肾脏中的一种以上功效。
7.根据权利要求5所述的应用，其特征在于：所述的药物是治疗2型糖尿病的药物。
8.根据权利要求5所述的应用，其特征在于：所述的药物还含有一种以上药学上可以接受的载体。
9.根据权利要求8所述的应用，其特征在于：所述的载体为缓释剂、赋形剂、填充剂、粘合剂、湿润剂、崩解剂、吸收促进剂、吸附载体、表面活性剂或润滑剂。
10.根据权利要求5所述的应用，其特征在于：所述的药物为片剂、颗粒剂或胶囊。
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