CN116158532A - 植物乳杆菌grx16在制备具有缓解胰岛素抵抗功能的食品、功能食品与药品中的应用 - Google Patents
植物乳杆菌grx16在制备具有缓解胰岛素抵抗功能的食品、功能食品与药品中的应用 Download PDFInfo
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- CN116158532A CN116158532A CN202310279050.7A CN202310279050A CN116158532A CN 116158532 A CN116158532 A CN 116158532A CN 202310279050 A CN202310279050 A CN 202310279050A CN 116158532 A CN116158532 A CN 116158532A
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- insulin resistance
- grx16
- lactobacillus plantarum
- insulin
- amylase
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Abstract
本发明涉及一种植物乳杆菌grx16在制备具有缓解胰岛素抵抗功能的食品、功能食品与药品中的应用，所述发酵乳杆菌grx08的保藏号为CGMCC No：10921。所述植物乳杆菌grx16能够缓解高碳水高脂饮食引起的血糖和胰岛素升高，降低胰岛素抵抗指数。经动物实验证明，植物乳杆菌grx16（CGMCC No：10921）能够增加调节胰岛素抵抗相关激素胰高血糖素样肽‑1和脂联素水平，抑制肠道淀粉酶和脂肪酶的活性和减少胰腺分泌淀粉酶，同时调节高脂饮食引起的炎症反应，缓解高碳水高脂饮食引起的血糖和胰岛素升高，降低胰岛素抵抗指数，进而缓解胰岛素抵抗。
Description
技术领域
本发明涉及一种植物乳杆菌grx16在制备具有缓解胰岛素抵抗功能的食品、功能食品与药品中的应用，属于生物技术领域。
背景技术
现代饮食结构的改变以及高脂、高碳水的饮食使得人体内脂肪和碳水化合物等代谢发生紊乱，导致肥胖的发生。胰岛素抵抗(IR)是肥胖与其大多数相关代谢紊乱(包括2型糖尿病，脂肪肝疾病，血脂异常和心血管疾病)之间的联系的基础，通常会在肥胖症的临床表现中表现出来。胰岛素抵抗是一个慢性非特异性炎症持续过程，IL-6等炎性因子影响胰岛素受体后信号转导。益生菌能够通过多种途径缓解胰岛素抵抗。在已有的临床和动物实验研究中，益生菌能够通过降低血糖、胰岛素、血脂、炎症因子等方式缓解胰岛素抵抗。此外，益生菌能够抑制肠道中消化酶类如淀粉酶和脂肪酶的活性，减少对淀粉酶和脂肪的消化吸收。本发明对已筛选获得的植物乳杆菌grx16进行体内缓解胰岛素抵抗功能的研究，为研发具有缓解胰岛素抵抗功能的食品、功能食品与药品打下基础。
发明内容
本发明目的是针对现有技术中存在的问题，提供一种植物乳杆菌grx16在制备具有缓解胰岛素抵抗功能的食品、功能食品与药品中的应用。
本发明的目的是通过以下技术方案实现的，植物乳杆菌(Lactobacillusplantarum)grx16在制备具有缓解胰岛素抵抗功能的食品、功能食品与药品中的应用，所述发酵乳杆菌grx08的保藏号为CGMCC No：10921。
所述植物乳杆菌grx16能够缓解高碳水高脂饮食引起的血糖和胰岛素升高，降低胰岛素抵抗指数。
所述植物乳杆菌grx16能够通过增加调节胰岛素抵抗相关激素胰高血糖素样肽-1和脂联素水平，抑制肠道淀粉酶和脂肪酶活性，同时抑制胰腺分泌淀粉酶，并调节高脂饮食引起的炎症反应来缓解胰岛素抵抗。
本发明方案科学合理，通过本发明，公开了植物乳杆菌grx16(CGMCC No：10921)在具有缓解胰岛素抵抗的食品、功能食品与药品中的应用。
经动物实验证明，植物乳杆菌grx16(CGMCC No：10921)能够增加调节胰岛素抵抗相关激素胰高血糖素样肽-1和脂联素水平，抑制肠道淀粉酶和脂肪酶的活性和减少胰腺分泌淀粉酶，同时调节高脂饮食引起的炎症反应，缓解高碳水高脂饮食引起的血糖和胰岛素升高，降低胰岛素抵抗指数，进而缓解胰岛素抵抗。
附图说明
图1胰岛素抵抗情况测定图；
*表示不同组别呈显著性差异(P＜0.05)。
图2调节胰岛素抵抗相关激素测定图；
*表示不同组别呈显著性差异(P＜0.05)。
图3大鼠肠道消化酶测定图；
*表示不同组别呈显著性差异(P＜0.05)。
图4大鼠胰腺消化酶测定图；
*表示不同组别呈显著性差异(P＜0.05)。
图5大鼠炎症因子测定图；
*表示不同组别呈显著性差异(P＜0.05)。
具体实施方式
中国专利ZL201510551144.0研究已经发现植物乳杆菌grx16(CGMCC No：10921)具有体外抗氧化和抑制α-葡萄糖苷酶活性的作用，本发明通过进一步研究发现植物乳杆菌grx16能够缓解高碳水高脂饮食引起的胰岛素抵抗。通过饲喂高碳水高脂饲料建立高碳水高脂饮食大鼠模型，灌胃植物乳杆菌grx16后，测定大鼠胰腺和食糜中淀粉酶(AMS)和脂肪酶(LPS)的活性，测定大鼠血清中血糖(GLU)，胰岛素(INS)、胰高血糖素样肽-1(GLP-1)、脂联素(ADPN)、肿瘤坏死因子-ɑ(TNF-ɑ)，白细胞介素-6(IL-6)和核因子κB(NF-κB)的含量，从而评价植物乳杆菌grx16缓解胰岛素抵抗的效果。
具体步骤
1灌胃样品制备
将保藏的植物乳杆菌grx16分别活化后，按3％的接种量接种到MRS液体培养基中，37℃培养18h后，4000×g离心10min后收集菌体。
植物乳杆菌grx16菌悬液：用生理盐水悬浮收集菌体并进行菌落计数，饲喂前用生理盐水稀释至1×109CFU/mL。
2动物分组及建模
SPF级SD雄性大鼠共15只，饲养在通风、透光和清洁卫生，室温为23.0±1.0℃，湿度为50±5％的动物房中，并用一周时间适应性饲喂高碳水基础饲料(面粉20％、米粉10％、玉米20％、鼓皮26％、豆料20％、鱼粉2％、骨粉2％)，适应有规律的12h白天/12h黑夜的循环，然后按各组间平均体重无显著差异重新分组，除空白组饲喂普通饲料外，其余各组通过饲喂4周高碳水高脂饲料(10％猪油、10％蛋黄粉、1％胆固醇和0.2％胆盐和78.8％基础饲料)建立高脂大鼠模型。造模成功后进行灌胃实验，时间为4周，大鼠分组及灌胃方式如表1所示。
表1高脂模型动物分组及处理
3混合益生菌对胰岛素抵抗的影响
胰岛素抵抗是指各种原因使胰岛素促进葡萄糖摄取和利用的效率下降，机体代偿性的分泌过多胰岛素产生高胰岛素血症，以维持血糖的稳定。常用胰岛素抵抗指数来反应胰岛素抵抗的程度。取大鼠血清，使用生化分析测定GLU浓度，ELISA试剂盒测定INS的含量，并计算胰岛素抵抗指数(HOMA-IR)，HOMA-IR＝空腹胰岛素×空腹血糖÷22.5。由图1可知，与空白组相比，模型组大鼠GLU和INS含量以及HOMA-IR指数均显著上升(P＜0.05)。与模型组相比，植物乳杆菌grx16干预显著降低了INS含量以及HOMA-IR指数(P＜0.05)，分别由51.07mU/L和18.86降至40.14mU/L和12.02，且均与空白组无显著性差异(P＞0.05)。GLU则较模型组有所下降，介于模型组和空白组之间，与模型组和空白组均无显著性差异(P＞0.05)。
4混合益生菌对调节胰岛素抵抗相关激素的影响
胰高血糖素样肽-1(GLP-1)是由肠道分泌的能够刺激胰岛素分泌，并通过中枢神经抑制食欲，达到降低血糖的作用，GLP-1水平降低会导致“肠促胰素效应”受损，是治疗2型糖尿病的重要靶点。脂联素(ADPN)则是由脂肪组织分泌的一种胰岛素增敏激素，能够改善胰岛素抵抗，并由一定的抗炎作用。取大鼠血清，使用ELISA试剂盒测定大鼠血清中GLP-1和ADPN的含量。由图2可知，与空白组相比，模型组大鼠GLP-1和ADPN的含量则显著降低(P＜0.05)。与模型组相比，植物乳杆菌grx16显著增加了GLP-1和ADPN的含量(P＜0.05)，分别由8.22pmol/L和7.96μg/mL增至10.04pmol/L和14.05μg/mL，其中是ADPN的含量恢复至空白组水平，与空白组物显著性差异(P＞0.05)。
5混合益生菌对大鼠消化酶活性的影响
肠道中淀粉酶(AMS)、脂肪酶(LPS)等消化酶的活性会影响大鼠对营养物质的吸收，胰腺则为分泌消化酶的主要器官。取大鼠胰腺和十二指肠内食糜，制备胰腺和食糜匀浆，分别使用淀粉酶活性测定试剂盒和脂肪酶活性测定试剂盒测定大鼠肠道和胰腺中AMS和LPS的活性。由图3可知，与空白组相比，模型组大鼠肠道内AMS和LPS的活性均显著升高(P＜0.05)。植物乳杆菌grx16干预后，相较于模型组，大鼠肠道内AMS和LPS的活性均显著降低(P＜0.05)，分别由0.61U/mg和8.02U/mg降至0.38U/mg和4.66U/mg，且均与空白组无显著性差异(P＞0.05)。由图4可知，与空白组相比，模型组胰腺中AMS和LPS的含量均无显著性差异(P＞0.05)。植物乳杆菌grx16干预后，大鼠胰腺AMS活性显著降低(P＜0.05)，由7.75U/mg降至0.63U/mg。
6混合益生菌对大鼠炎症因子的影响
细胞因子是机体防御系统的重要组成部分，既是免疫反应的产物，又可以增强免疫反应。TNF-α和IL-6具有广泛的生物学活性，被认为是反映炎性反应严重程度的重要指标，NF-κB作为一种多功能的细胞转录因子，在炎症反应、细胞凋亡等病理生理中起着重要作用。取大鼠血清，使用ELISA试剂盒测定大鼠血清中炎症因子TNF-ɑ，IL-6和NF-κB的含量。由图5可知，与空白组相比，模型组大鼠血清中TNF-ɑ，IL-6和NF-κB的含量均显著性增加(P＜0.05)。与模型组相比，植物乳杆菌grx16干预显著降低了大鼠血清中IL-6和NF-κB的含量(P＜0.05)，分别由146.96pg/mL和1302.25pg/mL降至111.12pg/mL和1032.01pg/mL，且均与空白组无显著性差异(P＞0.05)。
应用实施例
1植物乳杆菌grx16益生菌固体饮料的制备
植物乳杆菌grx16与低聚木糖、低聚果糖、桑叶提取物、木糖醇和天然果粉等中的一种或多种为原料，具体比例(质量百分数)为：植物乳杆菌grx16菌粉(活菌数1011CFU/g)5％-10％，低聚木糖15％-25％，低聚果糖10％-15％，桑叶提取物3％-5％，天然果粉15％-20％，木糖醇20％-25％。其中桑叶提取物为桑叶水体物(10：1)。原料混合后粉碎并过筛，充分混匀后包装。
2植物乳杆菌grx16发酵乳的制备
将全脂乳加热到50℃左右，加入6％蔗糖，经充分溶解后将全脂乳预热到60℃左右，在20-25MPa压力下进行均质后，95℃热处理5min，待冷却至40℃左右，按3％接种量接入植物乳杆菌grx16发酵剂，在37℃下发酵pH至4.2-4.5，冷却后于4℃贮藏。
3植物乳杆菌grx16发酵乳饮料的制备
350kg复原脱脂乳(含12％的脱脂奶粉)，经充分溶解后，95℃热处理5min，冷却至37-40℃，按3.5％接种量接入植物乳杆菌grx16发酵剂，在37℃下发酵24-26h左右，终点酸度控制在160-170°T左右，加入650kg经110℃热处理5s的糖、稳定剂混合溶液(该混合溶液的组成为12-14％蔗糖、0.1-0.15％单甘脂、0.1-0.15％蔗糖酯和0.4-0.8％果胶)，在
20 -25MPa压力下进行均质，冷却至15-20℃左右，采用无菌灌装，即得1000kg植物乳杆菌
grx16发酵乳饮料，4℃贮藏。
Claims (3)
1.植物乳杆菌grx16在制备具有缓解胰岛素抵抗功能的食品、功能食品与药品中的应用，所述发酵乳杆菌grx08的保藏号为CGMCC No：10921。
2.根据权利要求1所述的应用，其特征是，所述植物乳杆菌grx16能够缓解高碳水高脂饮食引起的血糖和胰岛素升高，降低胰岛素抵抗指数。
3.根据权利要求1所述的应用，其特征是，所述植物乳杆菌grx16能够通过增加调节胰岛素抵抗相关激素胰高血糖素样肽-1和脂联素水平，抑制肠道淀粉酶和脂肪酶活性，同时抑制胰腺分泌淀粉酶，并调节高脂饮食引起的炎症反应来缓解胰岛素抵抗。
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| CN105053202A (zh) * | 2015-09-01 | 2015-11-18 | 扬州大学 | 一种具有辅助降血糖功能的高品质发酵驼乳制备方法 |
| CN105132318A (zh) * | 2015-09-01 | 2015-12-09 | 扬州大学 | 植物乳杆菌Grx16及其应用 |
| CN114381395A (zh) * | 2021-12-30 | 2022-04-22 | 杭州康源食品科技有限公司 | 一种植物乳杆菌zjufn1及其应用 |
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| CN105053202A (zh) * | 2015-09-01 | 2015-11-18 | 扬州大学 | 一种具有辅助降血糖功能的高品质发酵驼乳制备方法 |
| CN105132318A (zh) * | 2015-09-01 | 2015-12-09 | 扬州大学 | 植物乳杆菌Grx16及其应用 |
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