CN110604790A - 益生菌联合铁皮石斛、余甘子及其提取物改善糖尿病及代谢综合征中的应用 - Google Patents
益生菌联合铁皮石斛、余甘子及其提取物改善糖尿病及代谢综合征中的应用 Download PDFInfo
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- CN110604790A CN110604790A CN201810633250.7A CN201810633250A CN110604790A CN 110604790 A CN110604790 A CN 110604790A CN 201810633250 A CN201810633250 A CN 201810633250A CN 110604790 A CN110604790 A CN 110604790A
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Abstract
本发明涉及生物技术领域，具体涉及益生菌联合铁皮石斛、余甘子及其提取物在调节血糖血脂，改善糖尿病及代谢综合征，减缓相关并发症的发生，进而维持机体代谢稳定。益生菌是一类对宿主有益的活性微生物，调节肠道菌群平衡，能产生确切健康功效从而改善宿主微生态平衡、发挥有益作用；余甘子具有调控血糖、增强机体免疫力等作用。本发明发现铁皮石斛及余甘子自身发挥降血糖降血脂作用，而且铁皮石斛及余甘子成分能为益生菌生长提供良好的环境，同时促进益生菌调节葡萄糖摄取，改善因单独服用益生菌产生的胃肠道刺激作用，从而使得益生菌和余甘子更好发挥降血糖作用，减少并发症的发生，可制成作为预防或治疗糖尿病及代谢综合征的药物组合物。因此，本发明为改善糖尿病、代谢综合征相关病症提供一种新用途。
Description
技术领域
本发明属于医药技术领域，具体涉及益生菌联合铁皮石斛、余甘子及其提取物改善糖尿病及代谢综合征相关并发症的新应用。
背景技术
糖尿病是一种能量代谢疾病，主要分为1型糖尿病(胰岛素依赖型糖尿病)和2型糖尿病(非胰岛素依赖型糖尿病)。目前全球约有3.66亿糖尿病患者，占世界人口的6.4％，其中2型糖尿病患者约占糖尿病患者总数的90～95％。二型糖尿病发病机制十分复杂，除了人体基因组、饮食习惯的改变或者体力活动减少而形成糖尿病，肠道菌群结构的改变与糖尿病的发生和发展存在一定的关系。肠道菌群作为环境因素能够显着影响宿主的营养吸收同时调节肠道免疫反应从而可能对宿主的病情有所影响。一旦肠道菌群失调，则引起人体代谢紊乱。
益生菌是一类对宿主有益的活性微生物，是定植于人体肠道、生殖系统内，能产生确切健康功效从而改善宿主微生态平衡、发挥有益作用的活性有益微生物的总称。干酪乳杆菌(Lactobacillus casei)能够耐受有机体的防御机制，其中包括口腔中的酶、胃液中低pH值和小肠的胆汁酸等，起到调节肠内菌群平衡、促进人体消化吸收等作用，具有高效降血压、降胆固醇，促进细胞分裂，产生抗体免疫，增强人体免疫及预防癌症和抑制肿瘤生长等功能；嗜酸乳杆菌(Lactobacillus acidophilus)通过分泌抗生物素类物质(嗜酸乳菌素(acidolin)、嗜酸杆菌素(acidophilin)、乳酸菌素(laetocidon)调整肠道菌群平衡，对致病微生物具有拮抗作用。双歧杆菌在肠道内发酵后，还可产生乳酸和醋酸，能提高钙、磷、铁的利用率，促进铁和维生素D 的吸收，产生维生素K及维生素B，还可以减少胆固醇的吸收，并能降低辐射对人体的伤害，可以预防腹泻，减少便秘，即双向调节，能起到预防和治疗各种肠道疾病的效果。副干酪乳杆菌(Lactobacillus paracasei) 作为一种益生菌，能促进机体微生物菌群和酶的平衡以及刺激特异性和非特异性的免疫机制，对革兰氏阳性菌和革兰氏阴性菌有明显抑制作用，抑菌谱广。植物乳杆菌(Lactobacillus plantarum)有一定的免疫调节作用，能有效改变各类型过敏，调整过敏体质，包括：过敏性鼻炎、结膜炎、异位性皮肤炎、荨麻疹、食物、花粉等各种过敏症状；嗜热链球菌(Streptococcus thermophiles)能帮助乳糖不耐受患者消化乳糖；能抑制轮状病毒的生长，能使艰难梭菌、鼠伤寒沙门氏菌、大肠杆菌等有有害菌的菌落直径减少约30％；同时能明显下调白细胞介素IL-6等炎症因子，对炎症有一定抑制作用。由于各类益生菌对其宿主营养、免疫、防病等具有显著的益生功效，越来越成为人们研究、开发、生产的焦点。
益生菌可以通过以下方式调节能量代谢，通过抑制脂质吸收、加速排泄，调节脂类摄入；促进胰高血糖素样肽-1释放和提高血管生成素样蛋白4水平，加大脂肪燃烧；促进粪便胆汁酸排出体外，加速脂质代谢过程；调节肠道激素，GLP-1和GLP-2是肠道L细胞分泌的肠促胰岛素，保护胰岛B细胞功能、调控脂肪摄取从而预防二型糖尿病发病。益生菌可以分泌抗菌物质，与其他病原菌竞争，从而抑制病原菌定植。同时益生菌参与肠道屏障、免疫炎症反应。
益生菌与糖尿病的发生密切相关，但是益生菌抗糖尿病的机制还没有完全被阐明，可能与减少氧化应激，调节免疫反应，减少炎症，调节肠道菌群相关。益生菌分泌的乳酸、乙酸和丁酸等物质能促进微量元素的吸收，如铁、钼、锰、铜、镁、硒、锗、锌、铬等。而在糖尿病的治疗中，铬是在醣类，脂质，蛋白质代谢中必需的微量元素，是胰岛素执行功能时的伴随因子，可以增加胰岛素的功能，可以增加胰岛素的结合能力，增加胰岛素的接受体数目，增加肝脏，肌肉，脂肪组织的葡萄糖运输。锌是脂质代谢中重要的辅助因子，有助于糖尿病人的脂质代谢，减少糖尿病人心血管疾病并发症的发生率。这是益生菌在糖尿病防治中所起的间接作用。
铁皮石斛(Dendrobium officinale Kimura et Migo)为兰科石斛属多年草本植物，多长于潮湿、温暖、半阴半阳的环境中，分布于福建、广西、云南等地，是我国传统的名贵中药材。现代中医药理认为，铁皮石斛味甘，性微寒。生津养胃；滋阴清热；润肺益肾；明目强腰。有研究表明铁皮石斛具有降血糖的功效，其能够调节胰岛α、β细胞分泌胰高血糖素和胰岛素而发挥降糖作用，同时其对胰岛β细胞具有修复作用，铁皮石斛还能抑制肾上腺素引起的肝糖原分解且促进肝糖原合成，从而达到控制血糖的作用。铁皮石斛还具有类似阿卡波糖的功效，能够抑制α-葡萄糖苷酶，延缓碳水化合物的消化和吸收。研究表明铁皮石斛具有增强机体免疫力的作用：铁皮石斛颗粒可促进荷瘤动物巨噬细胞的吞噬功能，增强T淋巴细胞的增殖和分化及NK细胞的活性，并能明显提高荷瘤动物的血清溶血素值，提示铁皮石斛无论是对非特异性免疫功能，或是特异性细胞免疫以及体液免疫功能，均有一定的提高作用。
余甘子(Phyllanthus emblica L.)为大戟科(Euphorbiaceae)叶下珠属(Phyllanthus)落叶灌木、小乔木果实，俗名滇橄榄、油甘子，初食酸涩，食后回甜生津，故名“余甘”，广泛分布于我国云南、广东、福建等省。在1998年余甘子被卫生部公布列入“药食同源”名单中，余甘子在防治糖尿病及其并发症方面具有良好的作用，具备开发利用价值。余甘子中含有多种对人体有益的物质：维生素C、黄酮类、鞣质类、皂苷以及多糖类物质，鞣质类成分具有良好的降血糖生物活性，其中主要为余甘子酸、油柑酸、葡萄糖没食子鞣质、没食子酸、并没食子酸、鞣料云实精、原诃子酸、诃子酸、诃子次酸、诃黎勒酸、余甘子酚等。具有免疫调节作用，保护胰岛β细胞，持续低血糖保护、改善胰岛素抵抗增加胰岛素敏感性、抗炎抗氧化等作用。
铁皮石斛颗粒可促进荷瘤动物巨噬细胞的吞噬功能，增强T淋巴细胞的增殖和分化及NK细胞的活性，并能明显提高荷瘤动物的血清溶血素值，提示铁皮石斛无论是对非特异性免疫功能，或是特异性细胞免疫以及体液免疫功能，均有一定的提高作用。余甘子具有持续低血糖保护、改善胰岛素抵抗增强胰岛素敏感性以及增强机体免疫力等作用，益生菌能调节肠道菌群平衡，改善宿主微生态平衡。铁皮石斛、余甘子和益生菌具有相辅相成的药用功效，但是到目前为止，未有报道发现三者共同使用能够相互促进改善糖尿病及代谢综合征，缓解其并发症发生的作用。本发明发现铁皮石斛和余甘子除了自身发挥降血糖降血脂作用，同时能作为益生元促进益生菌生长调节葡萄糖摄取，改善因单独服用益生菌产生的胃肠道刺激作用，从而达到更好的降血糖作用，减少并发症的发生。因此，本发明为改善糖尿病、代谢综合征相关病症提供一种新方法和新手段。
发明内容
一种益生菌、铁皮石斛和余甘子及其提取物的组合物在预防和治疗糖尿病及代谢综合征中的应用。
一种益生菌、铁皮石斛和余甘子及其提取物的组合物在预防和治疗糖尿病并发症中的应用。
一种益生菌、铁皮石斛和余甘子及其提取物的组合物在预防和治疗代谢综合征并发症中的应用。
所述益生菌，包括人体内已存在的肠道益生菌和/或经过人工改造过的应用的各类益生菌，包括：双歧杆菌(Bifidobacterium breve)、加氏乳杆菌(Lactobacillusgasseri)、长双歧杆菌(Bifidobacterium longum)、植物乳杆菌(Lactobacillusplantarum)、副干酪乳杆菌(Lactobacillus paracasei)、婴儿双歧杆菌(Bifidobacterium infantis)、嗜酸乳杆菌(Lactobacillus delbrueckii)、罗伊氏乳杆菌(Lactobacillus reuteri)、嗜热链球菌(Streptococcus thermophilus)、嗜酸乳杆菌(Lactobacillus acidophilus)、嗜酸乳杆菌La5(L. acidophilus La5)、双歧杆菌BB12(Bifidobacterium lactis Bb12)、干酪乳杆菌(Lactobacillus casei)、鼠李糖乳杆菌(Lactobacillus rhamnosus)、多枝杆菌(Bacteroides thetaiotaomicron)等，但不限于举例范围。
所述的铁皮石斛及其提取物，其特征在于石斛属提取物，其成分包括苯类及其衍生物：铁皮石斛素A、铁皮石斛素B、铁皮石斛素C、铁皮石斛素D、铁皮石斛素E、铁皮石斛素F、铁皮石斛素G、铁皮石斛素H、铁皮石斛素I、铁皮石斛素J、铁皮石斛素K、铁皮石斛素L、铁皮石斛素M、铁皮石斛素N、铁皮石斛素O、铁皮石斛素P、铁皮石斛素Q、4，4′-二羟基-3，5-二甲氧基联苄、3，4-二羟基-5，4′-二甲氧基联苄、 3′-羟基-3，4，5′-三甲氧基联苄、4，4′-二羟基-3，3′，5-三甲氧基联苄、3，4′-二羟基-5-甲氧基联苄、3′，4-二羟基-3，5′- 二甲氧基联苄、二氢白藜芦醇、dendromoniliside E、denbinobin、2，4，7-三羟基-9，10-二氢菲；酚类化合物： N-p-香豆酰酪胺，反-N-(4-羟基苯乙基)阿魏酸酰胺，二氢松柏醇二氢对羟基桂皮酸酯，二氢阿魏酸酪胺，对羟基苯丙酰酪胺，丁香酸，丁香醛，香草酸，对羟基苯丙酸，对羟基桂皮酸，阿魏酸，对羟基苯甲酸；木脂素类化合物：(+)-丁香脂素-O-β-D-吡喃葡萄糖苷，icariol A-2-4-O-β-D-glucopyranoside， (+)-lyoniresinol-3a-O-β-D-glucopyranoside，及裂异落叶松脂醇；内酯类化合物：钩状石斛素和洋地黄内酯；二氢黄酮类化合物：柚皮素和3′，5，5′，7-四羟基二氢黄酮；其他化合物：铁皮石斛素R，erigesideII，腺苷，尿苷，蔗糖，5-羟甲基糠醛，koaburaside，反式阿魏酸二十八烷基酯，对羟基反式肉桂酸三十烷基酯，对羟基顺式肉桂酸三十烷基酯，胡萝卜苷，β-谷甾醇，十六烷酸，十七烷，三十一烷醇，及十七烷酸；及各种单糖、寡糖、多糖类，但不限于举例范围。
所述的余甘子及其提取物，其特征在于大戟科叶下珠属提取物，其成分包括三十烷醇、三十烷酸、β- 香树脂酮、白桦脂酸酮、胡萝卜苷、羽扇豆醇乙酸酯、β-香树脂醇-3-棕榈酸酯、没食子酸、白桦脂酸、乌苏酸、齐墩果酸、槲皮素、芦丁、余甘子酸、油柑酸、葡萄糖没食子鞣质、并没食子酸、鞣料云实精、原诃子酸、诃子酸、诃子次酸、诃黎勒酸、余甘子酚、维生素C、黄酮类、鞣质类、皂苷以及多糖类物质等，但不限于举例范围。
所述的组合物，其特征在于由余甘子及其提取物各个成分单独或共同联合一种或多种益生菌。
所述的预防或治疗糖尿病、代谢综合征、及其并发症的组合物可通过口服或是被其他物质混合或包裹后导入机体。
所述余甘子及其提取物联合益生菌的组合物可作为活性成分制备预防或治疗糖尿病、代谢综合征、及其并发症，该组合物可制成片剂、粉剂、颗粒剂、胶囊、口服液、膏剂等多种形式，但不限于举例范围。
所述的代谢综合征是指人体的蛋白质、脂肪、碳水化合物等物质发生代谢紊乱，在临床上出现一系列综合征，即称代谢综合征。例如糖代谢紊乱时就出现糖耐量低减，导致糖尿病；脂肪代谢障碍时出现高脂备症，脂肪肝。肥胖症、高血粘稠度等，蛋白质代谢障碍，出现高尿酸血症(痛风)等。
所述代谢综合征并发症包括由代谢性疾病引发的急性并发症(恶心、呕吐、头晕、嗜睡、昏迷等)和慢性并发症(视网膜病变、肾病、神经病变等)。
一种益生菌、铁皮石斛、余甘子及其提取物的组合物联合降血脂药物的药物组合物在降血脂中的应用。其特征在于，所述的药物组合物还包括一种或多种降血脂药物包括：苯氧芳酸类：非诺贝特、吉非罗齐、苯扎贝特等；三羟甲基戊二酰-辅酶A还原酶抑制剂：洛伐他丁、辛伐他丁、普伐他丁等；烟酸类：氧甲吡嗪等；多不饱和脂肪酸类：橡胶种子油，月见草子，水飞蓟种子的油和海鱼的制剂等；泛硫乙胺；藻酸双酯钠以及其他降血脂药物：银杏类的药物等，但不限于举例范围。
一种益生菌、铁皮石斛、余甘子及其提取物的组合物联合降血糖药物的药物组合物在预防和治疗糖尿病及代谢综合征中的应用。其特征在于，所述的药物组合物还包括一种或多种治疗糖尿病药物，治疗糖尿病药物如胰岛素、胰岛素类似物、格列苯脲、格列喹酮、格列齐特、格列吡嗪、格列美脲、瑞格列奈、那格列奈、二甲双胍、苯乙双胍、环格列酮、罗格列酮、吡格列酮、利拉鲁肽、艾塞那肽、利司那肽、阿必鲁肽、阿卡波糖、伏格波糖、米格列醇、沙格列汀、西格列汀、维格列汀、利格列汀、阿格列汀、曲格列汀、达格列净、卡格列净、艾格列净、Ipragliflozin、Luseogliflozin以及Tofogliflozin等，但不限于举例范围。
实施例
本发明测试例中具体条件的实验方法通常按常规条件或按照商品制造厂商所建议的条件。未注明具体来源的试剂，为市场购买的常用试剂。
实施例1本实验以双歧杆菌、嗜热链球菌为例，铁皮石斛、余甘子及其提取物促进益生菌生长，抑制有害菌生长
复苏双歧杆菌、嗜热链球菌、大肠杆菌和金黄色葡萄球菌，双歧杆菌采用乳酸菌培养基、嗜热链球菌采用M17培养基，大肠杆菌采用LB培养基，金黄色葡萄球菌采用牛肉膏蛋白胨培养基培养。37℃过夜复苏后，按照1∶1000的比例加入培养基中，铁皮石斛和余甘子的比例为1∶1，混合均匀后分别给与不同浓度(0、0.02、0.1、0.2、1、2mg/mL)的余甘子提取物，每种浓度设置6个平行孔，在37℃生长8h后，采用OD600检测双歧杆菌、嗜热链球菌、大肠杆菌和金黄色葡萄球菌的生长情况。
表1铁皮石斛、余甘子提取物促对菌群生长的影响
*代表与无提取物时的对比，P＜0.05，**代表与无提取物时的对比，P＜0.01，***代表与无提取物时的对比， P＜0.001。
从上述实验结果可以得出，铁皮石斛、余甘子提取物能够促进益生菌的生长，对有害菌如大肠杆菌和金黄色葡萄球菌的生长有一定的抑制作用。
实施例2益生菌联合联合铁皮石斛、余甘子及其提取物控制体重增加
选用5周龄的雄性C57小鼠(青龙山动物实验中心，江苏南京)，体重20～24g，雄性，分为对照组 (采用高脂饲料喂养)、给药组1(铁皮石斛提取物)、给药组2(余甘子提取物)、给药组3(益生菌)和给药组4(铁皮石斛、余甘子和益生菌联合作用)，每组10只，雌雄各半。在喂高脂饲料的同时，分别口服给与不同的药物，每2周记录各组小鼠的体重，共12周，检测给药组小鼠的体重增长情况。
表2益生菌联合铁皮石斛、余甘子对提取物对小鼠体重增长的作用
*与对照组相比，P＜0.05；**与对照组相比，P＜0.01；***与对照组相比，P＜0.001
从实验结果可以得出，在高脂饮食的过程中，分别采用铁皮石斛、余甘子提取物和益生菌作用于小鼠，小鼠的体重增加得到有效的控制，具有显著性差异；而将益生菌联合铁皮石斛、余甘子提取物使用时，小鼠体重控制效果更好，与对照组相比具有极显著性差异。说明益生菌联合铁皮石斛、余甘子提取物共同作用可控制小鼠体重增加。
实施例3益生菌联合铁皮石斛、余甘子及其提取物降低高脂饮食诱导的糖尿病小鼠血糖
高脂饮食诱导的糖尿病小鼠口服糖耐量试验(OGTT)：选用5周龄的C57小鼠(青龙山动物实验中心，江苏南京)，体重20～24g，雄性，进行高脂饮食诱导小鼠肥胖。将成模的小鼠随机分为5组，分别为对照组，铁皮石斛组(100mg/kg)，余甘子组(100mg/kg)，益生菌组(100mg/kg)，铁皮石斛、余甘子提取物联合益生菌组(100mg/kg+100mg/kg+100mg/kg)，每组8只，每天灌胃给药，连续12周。实验结束时，实验前小鼠禁食不禁水12小时，灌胃给予葡萄糖(2g/kg)，断尾取血，测定给药后0，15，30，60，120 min测定血糖值。
表3益生菌联合铁皮石斛、余甘子及其提取物对高脂饮食诱导的糖尿病小鼠口服糖耐量影响
实验说明，通过益生菌联合铁皮石斛、余甘子提取物明显改善了高脂饮食诱导的糖尿病小鼠的口服糖耐量。
实施例4益生菌联合铁皮石斛、余甘子提取物降低高脂饮食诱导的肥胖小鼠血脂
高脂饮食诱导的肥胖小鼠口服糖耐量试验(OGTT)：选用5周龄的C57小鼠(青龙山动物实验中心，江苏南京)，体重20～24g，雄性，进行高脂饮食诱导小鼠肥胖。将成模的小鼠随机分为5组，分别为对照组，铁皮石斛组(100mg/kg)，余甘子组(100mg/kg)，益生菌组(100mg/kg)，铁皮石斛、余甘子提取物联合益生菌组(100mg/kg+100mg/kg+100mg/kg)，每组8只，每天灌胃给药，连续12周。实验结束时，取小鼠血清，分别检测小鼠空腹血清中的TG、TC、FFA和LDL-c的含量。
表4益生菌联合铁皮石斛、余甘子提取物对高脂饮食诱导的肥胖小鼠血脂影响
实验说明，益生菌联合铁皮石斛、余甘子提取物给药后，能降低小鼠体内的游离脂肪酸、甘油三酯、胆固醇和低密度脂蛋白，有效的改善了高脂饮食诱导的肥胖小鼠体内脂质代谢情况。
Claims (8)
1.一种益生菌、铁皮石斛、余甘子及其提取物的组合物在预防和治疗糖尿病及代谢综合征中的应用。
2.一种益生菌、铁皮石斛、余甘子及其提取物的组合物在预防和治疗糖尿病并发症中的应用。
3.一种益生菌、铁皮石斛、余甘子及其提取物的组合物在预防和治疗代谢综合征并发症中的应用。
4.权利要求1-3所述的组合物，其特征在于铁皮石斛、余甘子及其提取物各个成分单独或共同联合一种或多种益生菌。
5.权利要求1-3所述的预防或治疗糖尿病、代谢综合征、及其并发症的组合物可通过口服或是被其他物质混合或包裹后导入机体。
6.权利要求1-3所述铁皮石斛、余甘子及其提取物联合益生菌的组合物可作为活性成分制备预防或治疗糖尿病、代谢综合征、及其并发症，该组合物可制成片剂、粉剂、颗粒剂、胶囊、口服液、膏剂等多种形式，但不限于举例范围。
7.一种益生菌、铁皮石斛和余甘子及其提取物的组合物联合降血脂药物的药物组合物在降血脂中的应用。其特征在于，所述的药物组合物还包括一种或多种降血脂药物包括：苯氧芳酸类：非诺贝特、吉非罗齐、苯扎贝特等；三羟甲基戊二酰-辅酶A还原酶抑制剂：洛伐他丁、辛伐他丁、普伐他丁等；烟酸类：氧甲吡嗪等；多不饱和脂肪酸类：橡胶种子油，月见草子，水飞蓟种子的油和海鱼的制剂等；泛硫乙胺；藻酸双酯钠以及其他降血脂药物：银杏类的药物等，但不限于举例范围。
8.一种益生菌、铁皮石斛和余甘子及其提取物的组合物联合降血糖药物的药物组合物在预防和治疗糖尿病及代谢综合征中的应用。其特征在于，所述的药物组合物还包括一种或多种治疗糖尿病药物，治疗糖尿病药物如胰岛素、胰岛素类似物、格列苯脲、格列喹酮、格列齐特、格列吡嗪、格列美脲、瑞格列奈、那格列奈、二甲双胍、苯乙双胍、环格列酮、罗格列酮、吡格列酮、利拉鲁肽、艾塞那肽、利司那肽、阿必鲁肽、阿卡波糖、伏格波糖、米格列醇、沙格列汀、西格列汀、维格列汀、利格列汀、阿格列汀、曲格列汀、达格列净、卡格列净、艾格列净、Ipragliflozin、Luseogliflozin以及Tofogliflozin等，但不限于举例范围。
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