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干酪乳杆菌纳豆芽孢杆菌共培养上清液对抗生素相关性腹泻小鼠肠道微生态、黏膜屏障功能及免疫功能的影响
投稿时间:2019-08-23  修订日期:2019-10-08  点此下载全文
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作者单位E-mail
鲍小强 中国药科大学 2240131446@qq.com 
中文摘要:摘要:目的 探讨干酪乳杆菌(LC)、纳豆芽孢杆菌(BN)共培养上清液对抗生素相关性腹泻(AAD)小鼠肠道微生态、黏膜屏障功能及免疫功能的影响。方法 选取50只小鼠,其中40只建立AAD小鼠模型,随机分为模型组、LC组、BN组及共培养组,其余10只为对照组;LC组、BN组及共培养组分别灌胃LC、BN及共培养上清液,模型组和对照组灌胃等量生理盐水,连续给药4 d。观察干预期间小鼠一般情况,对比各组小鼠胸腺、脾脏体重比;观察近端结肠病变区组织病理学改变;对比各组盲肠内容物菌群多样性(Shannon)指数和丰富度(Chao)指数及优势菌群数量;检测并对比各组血清二胺氧化酶(DAO)水平及细菌异位率;检测肠黏膜中分泌型免疫球蛋白IgA(sIgA)及肠组织中白介素(IL)-2、IL-4水平。结果 模型组小鼠呈现精神状态不佳、采食量下降、粪便性状异常等改变,且随着时间延长而加重;干预后,各组小鼠上述状态均有所改善,其中共培养组恢复情况最佳;组织病理学结果显示,模型组肠壁组织结构破坏严重,组织绒毛脱落,可见纤维素样渗出、上皮细胞坏死、大量炎性细胞浸润;干预后上述病理变化均有所改善,其中共培养组改善效果最佳。与对照组相比,模型组胸腺、脾脏体重比、Shannon、Chao指数、乳杆菌数、双歧杆菌数、肠黏膜中sIgA、肠组织IL-2及IL-2/IL-4均较低,肠杆菌数、肠球菌数血清DAO、细菌异位率、肠组织IL-4均较高,差异均有统计学意义(P<0.05);与模型组相比,各干预组胸腺、脾脏体重比、Shannon、Chao指数、乳杆菌数、双歧杆菌数、肠黏膜中sIgA、肠组织IL-2及IL-2/IL-4均较高,差异均有统计学意义(P<0.05),其中共培养组高于LC组、BN组(P<0.05),LC组与BN组比较差异无统计学意义(P>0.05);与模型组相比,各干预组肠杆菌数、肠球菌数、血清DAO、细菌异位率、肠组织IL-4均较低,差异均有统计学意义(P<0.05),其中共培养组低于LC组、BN组(P<0.05),LC组与BN组比较差异无统计学意义(P>0.05);共培养组与对照组比较,血清DAO、细菌异位率、sIgA、IL-2、IL-4水平及IL-2/IL-4差异无统计学意义(P>0.05)。结论 LC和BN共培养上清液可有效调节AAD小鼠肠道微生态,改善肠黏膜屏障功能,提高肠道局部及整体免疫功能。
中文关键词:干酪乳杆菌  纳豆芽孢杆菌  共培养  抗生素相关性腹泻  肠道微生态  黏膜屏障  免疫功能
 
Effects of co-culture supernatant of Lactobacillus casei and Bacillus licheniformis on intestinal microecology, mucosal barrier function and immune function in mice with antibiotic-associated diarrhea
Abstract:Abstract: Objective To investigate the effects of co-culture supernatant of Lactobacillus casei (LC) and Bacillus natto (BN) on intestinal micro-ecology, mucosal barrier function and immune function in mice with antibiotic-associated diarrhea (AAD). Methods 50 mice were selected, 40 of them were established AAD mice model. They were randomly divided into model group, LC group, BN group and co-culture group. The remaining 10 mice were control group. LC group, BN group and co-culture group were given LC, BN and co-culture supernatant respectively. Model group and control group were given the same amount of saline for 4 days. The general conditions of mice during the intervention were observed and the weight ratios of thymus and spleen of mice in each group were compared. Histopathological changes of proximal colon lesions were observed. The red with the model group, the thymus, spleen weight ratio, Shannon, Chao index, Lactobacillus number, Bifidobacterium number, sIgA in intestinal mucosa, IL-2 and IL-2/IL-4 in intestinal tissue of the intervention groups were higher (P<0.05), and the co-culture group was higher than the LC group and the BN group (P<0.05). There was no significant difference between the LC group and the BN group (P>0.05). Compared with the model group, the number of Enterobacter, Enterococcus, serum DAO, bacterial ectopic rate and intestinal IL-4 in each intervention group were lower (P<0.05), and the co-culture group was lower than LC group and BN group (P<0.05). There was no significant difference between LC group and BN group (P>0.05). There were no significant differences in serum DAO, bacterial ectopic rate, sIgA, IL-2, IL-4 levels and IL-2/IL-4 levels between the co-culture group and the control group (P>0.05). Conclusion LC and BN co-culture supernatant can effectively regulate intestinal micro-ecology of AAD mice, improve intestinal mucosal barrier function, and improve intestinal and global immune function. Shannon index and Chao index of cecum contents and the number of dominant bacteria in each group were compared. The serum diamine oxidase (DAO) level and bacterial ectopic rate in each group were detected were and compared. The levels of secretory immunoglobulin IgA (sIgA) in intestinal mucosa and interleukin (IL) -2 and IL-4 in intestinal tissue were detected. Results The mice in the model group showed poor mental state, decreased feeding intake and abnormal stool characteristics, which were aggravated with the prolongation of time. After intervention, the above-mentioned state of mice in each group was improved, and the recovery of co-culture group was the best. Histopathological results showed that the intestinal wall of the model group was severely damaged and villus was shedding. Cellulose-like exudation, necrosis of epithelial cells and infiltration of inflammatory cells could be seen in the model group. The pathological changes mentioned above were improved after intervention, and the co-culture group had the best effect. Compared with the control group, the thymus and spleen weight ratio, Shannon index, Chao index, Lactobacillus number, Bifidobacterium number, sIgA in intestinal mucosa, IL-2 and IL-2/IL-4 in intestinal tissue of the model group were lower, while the number of Enterobacteria, Enterococcus number, serum DAO, bacterial ectopic rate and IL-4 in intestinal tissue were higher (P<0.05). Compa
keywords:Lactobacillus casei  Bacillus natto  Co-culture  Antibiotic-associated diarrhea  Intestinal microecology  Mucosal barrier  Immune function
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