CN116262125B - 保护胃黏膜、缓解胃炎的干酪乳杆菌l7-13及其应用 - Google Patents
保护胃黏膜、缓解胃炎的干酪乳杆菌l7-13及其应用 Download PDFInfo
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Abstract
本发明提供了一种保护胃黏膜、缓解胃炎的干酪乳杆菌L7‑13及其应用。本发明验证了干酪乳杆菌L7‑13在保护胃黏膜和缓解胃炎中的作用,并将干酪乳杆菌L7‑13与猴头菇和山楂的共提取物联用,具有预料不到的技术效果。本发明的技术方案可用于减少胃癌的发生,具有很好的应用前景。
Description
技术领域
本发明属于微生物制剂技术领域,具体涉及一种保护胃黏膜、缓解胃炎的干酪乳杆菌L7-13及其应用。
背景技术
胃炎(gastritis)是由众多原因引起的胃黏膜炎症,是最常见的消化系统疾病之一。按临床发病的缓急,一般可分为急性和慢性胃炎两大类型;按病因不同可分为幽门螺杆菌相关性胃炎、应激性胃炎、自身免疫性胃炎等。不同病因引起的胃炎病理改变亦不同,通常包括三个过程即上皮损伤、黏膜炎症反应和上皮再生。急性胃炎根据其病理改变又可分为单纯性、糜烂出血性、腐蚀性、化脓性胃炎等,慢性胃炎根据其病理改变可分为非萎缩性、萎缩性和特殊类型胃炎三大类。各型胃炎的诊断和鉴别诊断主要依据胃镜检查。
益生菌是通过定殖在人体内,改变宿主某一部位菌群组成的一类对宿主有益的活性微生物。通过调节宿主黏膜与系统免疫功能或通过调节肠道内菌群平衡,促进营养吸收保持肠道健康的作用,从而产生有利于健康作用的单微生物或组成明确的混合微生物。人体、动物体内有益的细菌或真菌主要有酵母菌、益生芽孢菌、丁酸梭菌、乳杆菌、双歧杆菌、放线菌等。酵母菌具有调节肠道平衡、促进饲料转化以及提高机体免疫功能等良好的益生特性,多作为饲料添加剂用于畜禽养殖。益生芽孢杆菌是一类对机体具有益生特性的、好氧或兼性厌氧、产芽孢的杆状细菌,为革兰氏染色阳性菌。丁酸梭菌是从健康人和动物肠道中分离出的一种厌氧的革兰氏阳性芽孢杆菌,具有维持肠道菌群平衡、增强机体免疫功能、生成营养物质、防治肠炎等良好的益生特性。乳酸菌指发酵糖类主要产物为乳酸的一类无芽孢、革兰氏染色阳性细菌的总称,是最常见的益生菌,已经被广泛应用,并认为对人和动物是安全的。双歧杆菌是人体内存在的一种生理性细菌,是人体有益菌中最值得重视和研究的一种,它与人体的健康密不可分。
专利CN201910301433.3公开了一种具有治疗慢性胃炎功能的多组分复合的益生菌,包括当归粉、木香粉、陈皮、酸枣粉、山药粉、双歧杆菌菌群、干酪乳杆菌菌群、植物乳杆菌菌群、发酵乳杆菌菌群、嗜酸乳杆菌、鼠李糖乳杆菌、嗜热糖链球菌、低聚果糖和非消化性聚糖,其所述的益生菌复合可以提升各个菌群中活菌的活性,有利于活菌繁殖,有利于提高药剂中活菌含量,达到更好的质量效果,更好的灭杀幽门螺杆菌,具有一定抑制胃酸的效果,有利于有益菌繁殖,促进胃粘膜修复,达到治疗慢性胃炎的作用。专利CN202110689217.8则公开了一种具有改善肠胃功效的药物组合物,所述组合物含有猴头菇20-60%,山楂10-30%,单糖1-20%,益生菌1-10%和药学上可接受的载体,其适用于防治各种原因引起的肠胃不适,如便秘、胃溃疡、胃炎等,并具有不刺激肠胃、通过和缓的改善肠道功能、不会引起血糖和血压的急速变化、施用方式灵活、无使用禁忌、可长期使用等优点。
目前针对保护胃黏膜、缓解胃炎的产品研究较少,因此,亟需提供一种保护胃黏膜、缓解胃炎效果更好的产品。
发明内容
针对以上问题,本发明提供了一种保护胃黏膜、缓解胃炎的干酪乳杆菌L7-13及其应用。本发明将干酪乳杆菌L7-13与猴头菇和山楂共提取物联用,具有较好的保护胃黏膜、缓解胃炎的效果。
为了实现上述发明目的,本发明的技术方案如下:
一方面,本发明提供了干酪乳杆菌L7-13在制备保护胃黏膜、缓解胃炎的产品中的应用。
具体地,所述的干酪乳杆菌L7-13保存于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为:CGMCC NO.3993。
具体地,所述的产品为保健品、食品或药物。
所述的保健品或食品可以保护胃黏膜。
所述的药物用于缓解胃炎。
进一步具体地,所述保健品的类型包括但不限于:药酒、胶囊、片剂、冲剂、茶制品、果汁、水果醋、口服液、软胶囊、颗粒、发酵奶制品、发酵谷制品、发酵豆制品、蜜膏、露剂、散剂、鲜汁或代餐粉。
所述保健品还包括保健品添加剂。
所述保健品添加剂包括但不限于:香精香料、着色剂、甜味剂、酸味剂、增鲜剂、乳化剂、增稠剂、防腐剂、抗氧剂或营养强化剂。
进一步具体地,所述的食品的类型包括但不限于:饼干、乳制品、代餐品、肉制品、酱汁、烘焙食品、酸奶、冰淇淋、发酵谷类基产品、果汁、米酒、糖果、糖浆、罐头食品、腌制品、调味品、豆制品、巧克力、馅料、茶制品或膨化食品。
所述的食品还包括食品添加剂。
所述的食品添加剂包括但不限于防腐剂、酸度调节剂、抗结剂、消泡剂、抗氧化剂、漂白剂、膨松剂、胶基糖果中基础剂物质、着色剂、护色剂、乳化剂、酶制剂、增味剂、面粉处理剂、被膜剂、水分保持剂、营养强化剂、防腐剂、稳定剂和凝固剂、甜味剂、增稠剂、食品用天然香料或食品用合成香料。
进一步具体地,所述的药物的剂型根据给药方式包括但不限于:经胃肠道给药剂型或非经胃肠道给药剂型。
所述的经胃肠道给药剂型包括但不限于散剂、片剂、颗粒剂、胶囊剂、溶液剂、乳剂、混悬剂或油剂。
所述的非经胃肠道给药剂型包括但不限于:注射给药剂型、呼吸道给药剂型、皮肤给药剂型、粘膜给药剂型或腔道给药剂型。
所述的注射给药剂型包括但不限于:静脉注射剂、肌内注射剂、皮下注射剂、皮内注射剂或腔内注射剂。
所述的呼吸道给药剂型包括但不限于喷雾剂、气雾剂或粉雾剂。
所述的皮肤给药剂型包括但不限于外用溶液剂、洗剂、搽剂、软膏剂、硬膏剂、糊剂或贴剂。
所述的粘膜给药剂型包括但不限于滴眼剂、滴鼻剂、眼用软膏剂、含漱剂、舌下片剂、粘贴片及贴膜剂。
所述的腔道给药剂型包括但不限于栓剂、气雾剂、泡腾片、滴剂及滴丸剂。
所述的药物还包括药学上可接受的辅料。
所述的药学上可接受的辅料包括但不限于溶剂、乳化剂、崩解剂、增溶剂、抗氧剂、pH调节剂、渗透压调节剂、抑菌剂、稀释剂、润湿剂、粘合剂或成膜剂。
所述的产品中还可以包括猴头菇和山楂的共提取物。
进一步具体地,所述的猴头菇和山楂的共提取物的制备方法包括以下步骤:
(1)取猴头菇和山楂切片,加入猴头菇和山楂总质量10-15倍的水,加热回流提取1-3次,每次30-60min;
(2)采用淀粉酶和糖化酶对步骤(1)的提取物进行酶解。
进一步具体地,所述的猴头菇和山楂的重量比为1-3:1-3,优选为1-2:1-2,进一步优选为1:1。
进一步具体地,所述的淀粉酶的浓度为400-600U/mL,所述的糖化酶的浓度为400-600U/mL。
进一步具体地,所述的淀粉酶与糖化酶的浓度比为1-2:1-2,优选为1:1。
进一步具体地,所述的酶解条件为:30-50℃条件下酶解2-4h。
具体地,所述的药物或食品或保健品中,干酪乳杆菌L7-13的含量为2.0×109-6.0×109CFU/g,优选为5.0×109CFU/g。
具体地,所述的干酪乳杆菌L7-13的培养基包括:溶剂为水,葡萄糖10-50g/L、大豆蛋白胨10-50g/L、磷酸氢二钾0.1-3g/L、柠檬酸三钠0.1-3g/L、柠檬酸铵0.05-0.2g/L、MgSO4·7H2O 0.1-1g/L、亚硝酸钠1-2g/L、吐温80 0.05-0.2mL/L。
进一步具体地,所述的干酪乳杆菌L7-13的培养基包括:溶剂为水,葡萄糖30g/L、大豆蛋白胨30g/L、磷酸氢二钾0.5g/L、柠檬酸三钠0.5g/L、柠檬酸铵0.1g/L、MgSO4·7H2O0.5g/L、亚硝酸钠1.5g/L、吐温80 0.1mL/L。
具体地,所述的组合物中,干酪乳杆菌和共提取物的重量比为1:1-20。
优选地,所述的组合物中,干酪乳杆菌和共提取物的重量比可以是1:1-10。
进一步优选地,所述的组合物中,干酪乳杆菌和共提取物的重量比可以是1:4-10。
更进一步地,所述的组合物中,干酪乳杆菌和共提取物的重量比可以是1:8。
另一方面,本发明提供了一种含益生菌的组合物。
所述的组合物中包括干酪乳杆菌L7-13,还包括猴头菇和山楂的共提取物;所述的干酪乳杆菌L7-13的保藏编号为CGMCC NO.3993。
所述的共提取物的制备方法包括以下步骤:
(1)取猴头菇和山楂切片,加入猴头菇和山楂总质量10-15倍的水,加热回流提取1-3次,每次30-60min;
(2)采用淀粉酶和糖化酶对步骤(1)的提取物进行酶解。
进一步具体地,所述的猴头菇和山楂的质量比为1-3:1-3,优选为1-2:1-2,进一步优选为1:1。
进一步具体地,所述的淀粉酶的浓度为400-600U/mL,所述的糖化酶的浓度为400-600U/mL。
进一步具体地,所述的淀粉酶与糖化酶的浓度比为1-2:1-2,优选为1:1。
进一步具体地,所述的酶解条件为:30-50℃条件下酶解2-4h。
具体地,所述的干酪乳杆菌L7-13的培养基包括:溶剂为水,葡萄糖10-50g/L、大豆蛋白胨10-50g/L、磷酸氢二钾0.1-3g/L、柠檬酸三钠0.1-3g/L、柠檬酸铵0.05-0.2g/L、MgSO4·7H2O 0.1-1g/L、亚硝酸钠1-2g/L、吐温80 0.05-0.2mL/L。
进一步具体地,所述的干酪乳杆菌L7-13的培养基包括:溶剂为水,葡萄糖30g/L、大豆蛋白胨30g/L、磷酸氢二钾0.5g/L、柠檬酸三钠0.5g/L、柠檬酸铵0.1g/L、MgSO4·7H2O0.5g/L、亚硝酸钠1.5g/L、吐温80 0.1mL/L。
具体地,所述的组合物中,干酪乳杆菌L7-13的含量为2.0-6.0×109CFU/g。
具体地,所述的组合物中,干酪乳杆菌和共提取物的重量比为1:1-20。
优选地,所述的组合物中,干酪乳杆菌和共提取物的重量比可以是1:1-10。
进一步优选地,所述的组合物中,干酪乳杆菌和共提取物的重量比可以是1:4-10。
更进一步地,所述的组合物中,干酪乳杆菌和共提取物的重量比可以是1:8。
再一方面,本发明提供了包含前述组合物的食品、保健品或药品。
所述的保健品或食品可以保护胃黏膜。
所述的药物用于缓解胃炎。
进一步具体地,所述保健品的类型包括但不限于:药酒、胶囊、片剂、冲剂、茶制品、果汁、水果醋、口服液、软胶囊、颗粒、发酵奶制品、发酵谷制品、发酵豆制品、蜜膏、露剂、散剂、鲜汁、代餐粉等。
所述保健品还包括保健品添加剂。
所述保健品添加剂包括但不限于:香精香料、着色剂、甜味剂、酸味剂、增鲜剂、乳化剂、增稠剂、防腐剂、抗氧剂、营养强化剂等。
进一步具体地,所述的食品的类型包括但不限于:饼干、乳制品、代餐品、肉制品、酱汁、烘焙食品、酸奶、冰淇淋、发酵谷类基产品、果汁、米酒、糖果、糖浆、罐头食品、腌制品、调味品、豆制品、巧克力、馅料、茶制品、膨化食品等。
所述的食品还包括食品添加剂。
所述的食品添加剂包括但不限于防腐剂、酸度调节剂、抗结剂、消泡剂、抗氧化剂、漂白剂、膨松剂、胶基糖果中基础剂物质、着色剂、护色剂、乳化剂、酶制剂、增味剂、面粉处理剂、被膜剂、水分保持剂、营养强化剂、防腐剂、稳定剂和凝固剂、甜味剂、增稠剂、食品用天然香料、食品用合成香料等。
进一步具体地,所述的药物的剂型根据给药方式包括但不限于:经胃肠道给药剂型或非经胃肠道给药剂型。
所述的经胃肠道给药剂型包括但不限于散剂、片剂、颗粒剂、胶囊剂、溶液剂、乳剂、混悬剂、油剂。
所述的非经胃肠道给药剂型包括但不限于:注射给药剂型、呼吸道给药剂型、皮肤给药剂型、粘膜给药剂型、腔道给药剂型。
所述的注射给药剂型包括但不限于:静脉注射剂、肌内注射剂、皮下注射剂、皮内注射剂、腔内注射剂。
所述的呼吸道给药剂型包括但不限于喷雾剂、气雾剂、粉雾剂。
所述的皮肤给药剂型包括但不限于外用溶液剂、洗剂、搽剂、软膏剂、硬膏剂、糊剂、贴剂。
所述的粘膜给药剂型包括但不限于滴眼剂、滴鼻剂、眼用软膏剂、含漱剂、舌下片剂、粘贴片及贴膜剂。
所述的腔道给药剂型包括但不限于栓剂、气雾剂、泡腾片、滴剂及滴丸剂。
所述的药物还包括药学上可接受的辅料。
所述的药学上可接受的辅料包括但不限于溶剂、乳化剂、崩解剂、增溶剂、抗氧剂、pH调节剂、渗透压调节剂、抑菌剂、稀释剂、润湿剂、粘合剂、成膜剂等。
本发明中所述的药物、食品、保健品的制备方法可采用本领域目前现有的以及未来研发出的相关制备方法。
与现有技术相比,本发明的积极和有益效果在于:
本发明将干酪乳杆菌L7-13与猴头菇和山楂的共提取物联用,具有较好的保护胃黏膜、缓解胃炎的效果,能够减少胃癌等疾病的发生。
具体实施方式
下面结合具体实施例,对本发明作进一步详细的阐述,下述实施例不用于限制本发明,仅用于说明本发明。以下实施例中所使用的实验方法如无特殊说明,实施例中未注明具体条件的实验方法,通常按照常规条件,下述实施例中所使用的材料、试剂等,如无特殊说明,均可从商业途径得到。
实验材料
干酪乳杆菌L7-13,保藏号CGMCC NO.3993,保存于中国微生物菌种保藏管理委员会普通微生物中心。所述的干酪乳杆菌L7-13详见专利CN201210578740.4。
淀粉酶为能水解淀粉分子链中的α-1,4-葡萄糖苷键,将淀粉链切断成为短链糊精和少量麦芽糖和葡萄糖,使淀粉粘度迅速下降的酶。通过购买获得,需要达到GB1886.174-2016国家标准。
糖化酶为以淀粉为底物,在一定条件下从淀粉的非还原性末端开始一次水解α-1,4-葡萄糖苷键产生葡萄糖的淀粉葡萄糖苷酶。通过购买获得,需要达到GB1886.174-2016国家标准。
实施例1一种保护胃黏膜、缓解胃炎的组合物
所述的组合物中包括干酪乳杆菌L7-13、猴头菇和山楂的共提取物,所述的干酪乳杆菌L7-13的含量为5.0×109CFU/g,所述的干酪乳杆菌L7-13与共提取物的重量比为1:8。
所述的干酪乳杆菌L7-13的培养方法如下:
培养基配方为:溶剂为水,葡萄糖30g/L,大豆蛋白胨30g/L,磷酸氢二钾0.5g/L、柠檬酸三钠0.5g/L、柠檬酸铵0.1g/L、MgSO4·7H2O 0.5g/L、亚硝酸钠1.5g/L、吐温80 0.1mL/L;pH5.0、35℃条件下培养36h,收集菌液制备成菌粉。
可以参照以下操作制备成菌粉:
(1)将格式干酪乳杆菌L7-13发酵液12000-18000rpm离心,操作时间在13小时内完成,收集菌泥,整个过程严格无菌操作;
(2)菌体包埋:加入包埋剂对菌体进行包埋,包埋剂与菌泥的比例为1:1,搅拌均匀,全过程严格无菌操作;
(3)冷冻干燥:进行预冻,将温度降至-40℃,然后升温至-20℃,保持30分钟,继续升温至-10℃,保持30分钟,之后以每增加5℃继续进行升温至26℃,每个升温过程大概需要3小时左右,并在每个温度下保持30分钟,冻干时间为30-48小时;
(4)粉碎:粉碎后,冻干粉为乳白色至棕色粉末或颗粒状,具有乳酸菌特殊气味;称量冻干粉重量,计算活菌数(CFU/g),保存。
所述的猴头菇和山楂的共提取物的制备方法如下:
(1)取猴头菇和山楂切片(重量比1:1),加入猴头菇和山楂总质量12倍的水,加热回流提取2次,每次45min;
(2)采用淀粉酶(500U/mL)和糖化酶(500U/mL),在40℃条件下对步骤(1)的提取物酶解3h。
实施例2一种保护胃黏膜、缓解胃炎的组合物
与实施例1的区别在于,干酪乳杆菌L7-13的含量为2.0×109CFU/g,所述的干酪乳杆菌L7-13与共提取物的重量比为1:4,制备共提取物时,猴头菇和山楂重量比为1:2。
实施例3一种保护胃黏膜、缓解胃炎的组合物
与实施例1的区别在于,干酪乳杆菌L7-13的含量为6.0×109CFU/g,所述的干酪乳杆菌L7-13与共提取物的重量比为1:10。制备共提取物时,猴头菇和山楂重量比为2:1。
对比例1-5
参照实施例1设置对比例1-5,具体如下:
实验例1胃粘膜保护效果检测
1、材料和方法
1.1样品:实施例1,对比例1-5制备的益生菌组合物。
1.2实验动物及环境:SPF级健康SD种雄性大鼠,体重170.2±5.7g,由广东省医学实验动物中心繁殖,实验动物生产许可证号:SCXK(粤)2008-0002,实验动物质量合格证号:0081569。动物实验室为屏障系统,使用许可证号:SYXK(桂)2007-0003。实验动物室温度:22-25℃,相对湿度:55-70%。
1.3剂量选择与受试物给予方式:每组10只大鼠,将上述实施例1、对比例1-5制备的产品按照1333mg/kg BW(133.3mg/mL浓度)灌胃,灌胃体积为1.0mL/100g BW,同时设置正常对照组及模型对照组,予以等体积的纯水,每天灌胃一次,连续灌胃30天。
1.4主要仪器与试剂
仪器:解剖镜、游标卡、解剖器械。
试剂:无水乙醇、生理盐水、10%甲醛。
1.5实验方法:依据《保健食品检验与评价技术规范-2003》中对胃粘膜损伤有辅助保护功能检验方法-急性胃粘膜损伤模型法进行实验。大鼠灌胃受试物30天,解剖前动物禁食不禁水24时。实验结束当天灌胃受试物1h后再灌胃无水乙醇1.0mL只,1h后处死动物,取胃置于10%甲醛溶液中固定20分钟后,于立体显微镜下观察粘膜的损伤情况并用游标卡尺测定胃粘膜损伤的程度。
游标卡尺测定胃粘膜损伤的程度:将固定后的胃沿胃大弯剪开,洗净胃内容物,将胃粘膜展开,胃粘膜损伤表现为腺胃区粘膜的条状充血或弥漫性出血,用游标卡尺测量出充血或弥漫性出血条带的面积(精确到0.01mm),以充血带和弥漫性出血的总面积作为损伤程度的评价指标。
1.6试验数据处理:以方差分析进行统计处理。
1.7结果判定:实验组与阴性对照组进行比较,胃粘膜损伤面积明显小于对照组,且统计学处理差异有显著性,则可判定该受试样品对胃粘膜损伤有辅助保护功能动物实验结果阳性。
2、实验结果
2.1样品对大鼠体重的影响
由下表1可只,试验前各组大鼠体重无明显差异,试验结束时各组大鼠的体重及增重量比较,差异均没有显著性(P>0.05),表明该样品对受试大鼠的体重增长无明显影响。
表1.试验前后大鼠体重变化
2.2样品对大鼠的胃粘膜损伤程度的影响
由下表2可知,本发明所述的益生菌组合物对胃粘膜损伤具有更好的辅助保护功能(P<0.05),且本发明所述的益生菌组合物具有更好的协同增效的作用。
表2.各组大鼠急性胃粘膜损伤结果
实验例2
1、受试对象
1.1受试者纳入标准:符合慢性浅表性胃炎诊断标准且经胃镜筛选确诊为浅表性胃炎的18-60岁的自愿受试者。慢性浅表性胃炎诊断标准:病程迁延,有不同程度的消化不良、上腹痛、烧心、嗳气、反酸、腹胀等临床症状,可有上腹部位轻度压痛。符合慢性浅表性胃炎纤维胃镜诊断标准及活体组织检查诊断标准,排除胃溃疡患者。
1.2受试者排除标准:(1)年龄在18岁以下或60岁以上者,妊娠或哺乳期妇女及对本样品过敏者;(2)继发性慢性胃炎患者;(3)合并有心、脑血管、肝、肾和造血系统等严重全身性疾病及精神病患者;(4)经常用药、嗜酒、大量吸烟者;(5)有消化系统溃疡的病人,症状、体征分级为重症者;(6)正在服用其它治疗药物或接受其它治疗者;(7)短期内服用与受试功能有关的物品,影响到对结果的判断者:(6)未按规定服用受试样品无法判断功效,或资料不全等影响功效或安全性判断者。
2、试验设计与分组
采用自身和组间两种对照设计。按受试者的症状轻重随机分为试食组和对照组,尽可能考虑影响结果的主要因素如年龄、性别、病程等,进行均衡性检验,以保证组间的可比性。每组最终进入有效统计的受试者不少于50例。
3、食用剂量及时间
试食组试食样品每人每天早晚各服食1次,每次1袋,连续服用45天;对照组服用安慰剂,用法、用量同试验组。
4、主要仪器
心电图、X线透视机、B超扫描仪、生化分析仪、血球计数仪、血压计等。
5、观察指标
5.1安全性指标
5.1.1一般情况:包括精神状况、睡眠、饮食、大小便等,每天由受试者记录。
5.1.2血常规检查:应用全自动血球计数仪检测血常规指标,试验前、试验结束各做一次。
5.1.3大、小便常规检查:试验前、试验结束各做一次。
5.1.4血压和肝、肾功能检查:用血压计测量血压,应用全自动生化分析仪测定血液主要生化指标,试验前、试验结束各做一次。
5.1.5胸透、心电图、腹部B超:在试验开始前进行一次胸部透视心电图和腹部B超检查。
5.2功能性指标
5.2.1症状观察:详细询问病史,主要临床症状:胃痛、喛气、反酸、腹胀、食欲不振、少食等,按症状轻重(重症3分、中度2分、轻度1分)统计积分,症状轻重分级见表3。
表3.症状轻重分级
5.2.2胃镜复査及体征观察
试食组和对照组全部受试者进行胃镜复查,比较试食试验前后的改变。
剑突下压痛程度检查,根据疼痛程度分为轻(1分)、中(2分)、重(3分):(1)轻度:用力时才出现疼痛,压痛轻微1分;(2)中度:用力即出现疼痛,但疼痛尚能忍受,压痛明显2分;(3)重度:稍微用力即出现疼痛,疼痛不能忍受,压痛剧烈3分。
6、数据处理
统计症状和体征积分值。凡自身对照资料可以采用配对t检验,两组均数比较采用成组t检验,后者需进行方差齐性检验,对非正态分布或方差不齐的数据进行适当的变量转换,待满足正态方差齐后,用转换的数据进行t检验;若转换数据仍不能满足正态方差齐要求,改用t’检验或秩和检验;但变异系数太大(如CV>50%)的资料应用秩和检验。
7、结果判定
试食前后试食组自身比较及试食后试食组与对照组组间比较,临床症状、体征积分明显减少,胃镜复查结果有改善或不加重,且症状、体征改善率较对照组有明显増加,差异有显著性,可判定该受试样品具有对胃粘膜损伤辅助保护功能。
8、结果
8.1一般情况:
试验前,两组各有60例志愿者参加受试;试验终末,失访6例,失访率5.0%。试食前,试食组与对照组人群的年龄、精神状况、睡眠情况、饮食情况等基本一致,血压、血、尿、便常规及血液生化指标等检查均未见异常;两组人群的胸部透视、心电图及腹部B超检查结果均未见明显异常。腹部症状、体征积分均没有显著性差异(P>0.05),见表4。
表4.试食前两组人群腹部症状、体征积分比较
8.2对慢性浅表性胃炎人群功能性指标的影响
表5.试食前后两组人群的腹部症状积分比较
组别 | 人数 | 试食前总积分 | 试食后总积分 | 总积分下降数值 | P值(自身) |
对照组 | 56 | 3.18±0.58 | 2.99±0.52 | 0.19±0.55 | >0.05 |
试食组 | 58 | 3.18±1.04 | 0.73±0.83 | 2.46±1.07 | <0.01 |
P值(组间) | >0.05 | <0.01 | <0.01 | — |
表6.试食前后两组人群的腹部体征积分比较
表7.样品对慢性浅表性胃炎人群腹部症状、体征改善情况
注:*胃镜每组只随机复查30例,故改善率以30例计算。
从表5-表7可见,试食后,试食组人群的腹部症状总积分和体征总积分均明显低于对照组,且试验组与对照组间的差异均有显著性(P<0.01);试验组试食前后症状总积分、体征总积分自身比较也都具有显著性差异(P<0.01);试验组的症状总积分及体征总积分的下降值均明显大于对照组,差异均具有显著性(P<0.01)。试验组的各个腹部症状和体征的改善率及总改善率均明显高于对照组,差异具有显著性(P<0.01)。结果表明,该样品具有减轻试食者的腹部症状和体征的作用。
8.3样品对安全性指标的影响
8.3.1样品对血压、心率的影响
试食前后,试验组与对照组人群的收缩压和舒张压均在正常值范围内,两组比较均无显著性差异(P>0.05),见表8,表明该样品对受试者的血压和心率无不良影响。
表8.试验前后两组人群血压(mmHg)、心率(次/分)的测量结果
8.3.2样品对尿、便常规的影响
试食前后,试验组与对照组人群的尿液酸碱度、透明度、颜色和尿沉渣镜检均未见异常,蛋白均为阴性。两组的粪便颜色、性状和镜检均未见异常,见表9、表10,表明该样本对受试者的大、小便常规无不良影响。
表9.试验前后两组人群大便常规的检验结果
表10.试验前后两组人群尿常规的检验结果
8.3.3样品对血常规、血清生化指标的影响
试食前后,两组人群的血红细胞数、白细胞数、血红蛋白含量和血小板数均在正常值范围内,试食组自身前后比较及与对照组的组间比较均无显著性差异(P>0.05),见表11,表明该样品对受试者的血常规项目无不良影响。试食前后,两组人群的血液生化指标检查结果均在正常值范围内,试食组自身前后比较及与对照组的组间比较均无显著性差异(P>0.05),见表12,表明该样品对受试者的肝、肾功能无不良影响。
表11.试验前后两组人群血常规的检验结果
表12.试验前后两组人群血液生化的检验结果/>
8.3.4样品的不良反应
试验过程中试食样品者均未出现恶心、胀气、腹泻及过敏等不良反应。
9、小结
9.1符合本试验受试入选标准的慢性浅表性胃炎人群114例分为两组(对照组56例:男27例,女29例;试验组58例:男29例,女29例),对照组和试验组分别服用安慰剂和受试样品,连续服用45天后,对照组和试验组的腹部症状、体征总积分均有降低,试验组尤为明显,其下降值与对照组的下降值比较差异均具有显著性(P<0.01);试验组的腹部症状、体征总积分自身前后比较均有显著性差异(P<0.01);试验组腹部症状和体征改善率分别为93.1%和69.0%,显著高于对照组的28.6%和5.4%(P<0.01)。由此可见,该样品具有对胃粘膜损伤辅助保护功能。
9.2试验前后,试食样品组人群的血红细胞数、白细胞数、血红蛋白、大小便常规、血清总蛋白、白蛋白、谷草转氨酶、谷丙转氨酶、尿素氮、肌酐、血糖等各项检查结果均在正常值范围内,与对照组的差异均无显著性(P>0.05),表明该样品对受试者健康无不良影响。
9.3试验过程中试食样品者均未出现恶心、胀气、腹泻及过敏等不良反应。
由此可见,本发明所述样品具有对胃粘膜损伤辅助保护功能,并对受试人群健康无损害作用。
以上所述实施例仅表达了本发明的几种实施方式,其描述较为具体和详细,但并不能因此而理解为对本发明专利范围的限制。应当指出的是,对于本领域的普通技术人员来说,在不脱离本发明构思的前提下,还可以做出若干变形和改进,这些都属于本发明的保护范围。因此,本发明专利的保护范围应以所附权利要求为准。
Claims (7)
1.干酪乳杆菌L7-13在制备缓解胃黏膜损伤的药物中的应用,其特征在于,所述的干酪乳杆菌L7-13保藏编号为CGMCC NO.3993,干酪乳杆菌L7-13的含量为2.0×109-6.0×109CFU/g;
所述的药物中还包括猴头菇和山楂的共提取物;所述的干酪乳杆菌L7-13和共提取物的重量比为1:1-20;
所述的共提取物的制备方法包括以下步骤:
(1)取猴头菇和山楂切片,加入猴头菇和山楂总质量10-15倍的水,加热回流提取1-3次,每次30-60min;
(2)采用淀粉酶和糖化酶对步骤(1)的提取物进行酶解;
所述的猴头菇和山楂的质量比为1-3:1-3;
所述的淀粉酶的浓度为400-600 U/mL,所述的糖化酶的浓度为400-600 U/mL;
所述的干酪乳杆菌L7-13的培养基由以下成分组成:溶剂为水,葡萄糖10-50g/L、大豆蛋白胨10-50g/L、磷酸氢二钾0.1-3g/L、柠檬酸三钠0.1-3g/L、柠檬酸铵0.05-0.2g/L、MgSO4·7H2O 0.1-1g/L、亚硝酸钠1-2g/L、吐温80 0.05-0.2mL/L。
2.干酪乳杆菌L7-13在制备保护胃黏膜的食品或保健品中的应用,其特征在于,所述的干酪乳杆菌L7-13保藏编号为CGMCC NO.3993,干酪乳杆菌L7-13的含量为2.0×109-6.0×109CFU/g;
所述的药物中还包括猴头菇和山楂的共提取物;所述的干酪乳杆菌L7-13和共提取物的重量比为1:1-20;
所述的共提取物的制备方法包括以下步骤:
(1)取猴头菇和山楂切片,加入猴头菇和山楂总质量10-15倍的水,加热回流提取1-3次,每次30-60min;
(2)采用淀粉酶和糖化酶对步骤(1)的提取物进行酶解;
所述的猴头菇和山楂的质量比为1-3:1-3;
所述的淀粉酶的浓度为400-600 U/mL,所述的糖化酶的浓度为400-600 U/mL;
所述的干酪乳杆菌L7-13的培养基由以下成分组成:溶剂为水,葡萄糖10-50g/L、大豆蛋白胨10-50g/L、磷酸氢二钾0.1-3g/L、柠檬酸三钠0.1-3g/L、柠檬酸铵0.05-0.2g/L、MgSO4·7H2O 0.1-1g/L、亚硝酸钠1-2g/L、吐温80 0.05-0.2mL/L。
3.根据权利要求1或2所述的应用,其特征在于,所述的干酪乳杆菌L7-13的培养基由以下成分组成:溶剂为水,葡萄糖30g/L、大豆蛋白胨30g/L、磷酸氢二钾0.5g/L、柠檬酸三钠0.5g/L、柠檬酸铵0.1g/L、MgSO4·7H2O 0.5g/L、亚硝酸钠1.5g/L、吐温80 0.1mL/L。
4.根据权利要求3所述的应用,其特征在于,所述的淀粉酶与糖化酶的浓度比为1:1。
5.根据权利要求4所述的应用,其特征在于,所述的干酪乳杆菌L7-13的含量为5.0×109CFU/g。
6.一种含益生菌的组合物,其特征在于,包括干酪乳杆菌L7-13,还包括猴头菇和山楂的共提取物;所述的干酪乳杆菌L7-13的保藏编号为CGMCC NO.3993;干酪乳杆菌L7-13的含量为2.0×109-6.0×109CFU/g;
所述的干酪乳杆菌L7-13和共提取物的重量比为1:1-20;
所述的共提取物的制备方法包括以下步骤:
(1)取猴头菇和山楂切片,加入猴头菇和山楂总质量10-15倍的水,加热回流提取1-3次,每次30-60min;
(2)采用淀粉酶和糖化酶对步骤(1)的提取物进行酶解;
所述的猴头菇和山楂的质量比为1-3:1-3;
所述的淀粉酶的浓度为400-600 U/mL,所述的糖化酶的浓度为400-600 U/mL;
所述的干酪乳杆菌L7-13的培养基由以下成分组成:溶剂为水,葡萄糖10-50g/L、大豆蛋白胨10-50g/L、磷酸氢二钾0.1-3g/L、柠檬酸三钠0.1-3g/L、柠檬酸铵0.05-0.2g/L、MgSO4·7H2O 0.1-1g/L、亚硝酸钠1-2g/L、吐温80 0.05-0.2mL/L。
7.包含权利要求6所述的组合物的食品、保健品或药品。
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