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母乳中乳酸菌分离株的有趣益生菌特性;从细菌素到改善炎症性肠病。

Interesting probiotic traits of mother's milk Lactobacillus isolates; from bacteriocin to inflammatory bowel disease improvement.

机构信息

Department of Microbiology, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran.

Department of Virology, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran.

出版信息

Microb Pathog. 2021 Sep;158:104998. doi: 10.1016/j.micpath.2021.104998. Epub 2021 May 24.

Abstract

AIMS AND BACKGROUND

Lactobacillus spp. are an important element in breast milk. This component has a beneficial effect on the composition of the intestinal microflora and the intestinal immune system. The aim of this study was to isolate and identify Lactobacillus strains in breast milk and evaluate some of their probiotic properties, such as presence of bacteriocin genes, adhesion to HT-29 cell line, competition with enteropathogens in cell culture, and effect on serum level of lipids and digestive enzymes, and mice model of inflammatory bowel disease (IBD).

MATERIALS AND METHODS

A total of 323 lactic acid bacteria (LAB) were isolated from breast milk samples of healthy mothers with the age ranges from 21 to 45 years old. These isolates were subjected to phenotypic and molecular experiments. The frequency of bacteriocin genes was determined by polymerase chain reaction (PCR). Adhesion of Lactobacillus isolates to HT-29 cells was measured based on the number of attached bacterial cells in 20 fields of the light microscopy. Competition test was done by colony count and real-time PCR procedures. Five strongly adhesive Lactobacillus strains were selected and administered orally to the treatment groups. After 8 days, the serum level of digestive enzymes and improvement in induced IBD, and after 14 days, the serum level of lipids (triglycerides, total cholesterol, HDL, and LDL) in treated mice were surveyed compared to the control groups.

RESULTS

Based on the phenotypic and molecular experiments, L. casei, L. plantarum, L. rhamnosus, and L. acidophilus strains were isolated and identified in the breast milk samples. The highest frequency of bacteriocin genes belonged to Plantaricin B (100%), followed by Plantaricin D (84.7%), Plantaricin G (84.7%), and Plantaricin EF (54.3%). Also, 71.8% of the isolates were strongly adhesive, 21.8% were non-adhesive, and 6.4% were adhesive. Lactobacillus strains had a significant effect on the displacement of enteropathogens. The in vitro cholesterol-removing ability of L. casei (L1), L. casei (L2), L. casei (L3), L. plantarum (L4), and L. rhamnosus (L5) was 3.5, 31.5, 21.3, 18.7, and 27.3%, respectively. The serum level of total cholesterol in the L. plantarum (L4) group as well as LDL in the L. casei (L3) (p = .0108) and L. rhamnosus (L5) (p = .0206) groups decreased significantly compared to the control group. The serum level of lipase increased in all the treatment groups compared to the control group, which was significant in the L. plantarum (L4) group (p = .0390). Disease activity index (DAI) scores were improved significantly in L. casei (L3) group compared to the IBD control group (p < .0001).

CONCLUSION

It could be concluded that lactobacilli strains isolated from the breast milk samples had good probiotic properties, such as presence of bacteriocin genes, attaching to enterocyte-like HT-29 cells, competing with intestinal pathogens, lowering cholesterol, and improving IBD. Thus, after further studies, they could be considered as probiotic strains.

摘要

目的和背景

乳杆菌属是母乳中的重要组成部分。这一成分对肠道微生物群组成和肠道免疫系统有有益影响。本研究的目的是从母乳中分离和鉴定乳杆菌菌株,并评估其一些益生菌特性,如存在细菌素基因、黏附于 HT-29 细胞系、在细胞培养中与肠道病原体竞争,以及对血清脂质和消化酶水平的影响,以及在炎症性肠病(IBD)的小鼠模型中的作用。

材料和方法

从 323 位年龄在 21 至 45 岁之间的健康母亲的母乳样本中分离出 323 株乳酸杆菌(LAB)。这些分离株进行了表型和分子实验。通过聚合酶链反应(PCR)确定细菌素基因的频率。根据光镜下 20 个视野中附着的细菌细胞数量来测量乳杆菌分离株对 HT-29 细胞的黏附。通过菌落计数和实时 PCR 程序进行竞争试验。选择 5 株黏附性较强的乳杆菌菌株进行口服治疗。8 天后,与对照组相比,检测治疗组的血清消化酶水平和诱导的 IBD 改善情况,14 天后,检测血清脂质(甘油三酯、总胆固醇、HDL 和 LDL)水平。

结果

根据表型和分子实验,从母乳样本中分离和鉴定出了 L. casei、L. plantarum、L. rhamnosus 和 L. acidophilus 菌株。细菌素基因的最高频率属于 Plantaricin B(100%),其次是 Plantaricin D(84.7%)、Plantaricin G(84.7%)和 Plantaricin EF(54.3%)。此外,71.8%的分离株具有较强的黏附性,21.8%的分离株不具有黏附性,6.4%的分离株具有黏附性。乳杆菌菌株对肠道病原体的置换有显著影响。L. casei(L1)、L. casei(L2)、L. casei(L3)、L. plantarum(L4)和 L. rhamnosus(L5)的胆固醇去除能力分别为 3.5%、31.5%、21.3%、18.7%和 27.3%。L. plantarum(L4)组的总胆固醇血清水平以及 L. casei(L3)组的 LDL(p =.0108)和 L. rhamnosus(L5)组(p =.0206)的血清水平与对照组相比显著降低。与对照组相比,所有治疗组的血清脂肪酶水平均升高,其中 L. plantarum(L4)组的升高具有统计学意义(p =.0390)。与 IBD 对照组相比,L. casei(L3)组的疾病活动指数(DAI)评分显著改善(p <.0001)。

结论

从母乳样本中分离出的乳杆菌菌株具有良好的益生菌特性,如存在细菌素基因、黏附于肠上皮样 HT-29 细胞、与肠道病原体竞争、降低胆固醇和改善 IBD。因此,在进一步研究后,它们可以被考虑作为益生菌菌株。

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