Lactobacillus acidophilus Supplementation Restores Gut Epithelial Integrities and Barrier Functions in Non-specific Diarrhea

Hossain Ferdaus Mohd Altaf, Sumaiya Farzana, K. Parvez, Z. Eman, Atif Muhammad, U. Nazim, Zinnah Ali, Khan IMDAD ULLAH, Rahman Masudur
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Abstract

Objectives: The study was aimed to evaluate the roles of Lactobacillus acidophilus supplementation in maintaining intestinal epithelial integrities, tight junction proteins, and adhesion molecules in non-specific diarrhea. Methods: In this study, we used the pre-weaned BL/6 pups (3 weeks of age, same litters) as a model animal. We supplied the non-sterilized and poor-quality water to experimental pups (n=7) to develop non-specific diarrheic symptoms. Then diarrheic pups were supplemented with Lactobacillus acidophilus (LA) for three consecutive weeks. The control group (n=5) was supplied with sterilized water and no LA. The sampling and analysis were performed on day 0, day 7, day 14, and day 21. The expressions of pro-inflammatory cytokines (IL-6, TNF-a) and tight junction proteins (TJPs) of gut mucosa were determined using qRT-PCR. And the serum cytokines level was screened through sandwich ELISA. Results: The intestinal cytoskeletal integrity becomes disrupted and characterized by lower ZO-1, Occludin, Claudin-1, Claudin-5, and JAM mRNA expressions upon real-time qRT-PCR. However, Claudin-4 was found to be not affected and illustrated with a higher expression like control pups. Interestingly, supplementing Lactobacillus acidophilus was found to maintain gut integrity and effectively reduce diarrheic symptoms. Like the control pups, the Lactobacillus acidophilus supplemented pups exhibited a higher expression of gut epithelial TJPs and adhesion molecules. Moreover, the diseased pups produced significantly increased IL-6, and TNF-α production in blood serum, compared to control BL/6 pups. Conclusion: We concluded that L. acidophilus supplementation might orchestrate the equilibrium of gut health and immunity against non-specific diarrhea.
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补充嗜酸乳杆菌可恢复非特异性腹泻患者肠道上皮完整性和屏障功能
目的:本研究旨在评估补充嗜酸乳杆菌在非特异性腹泻中维持肠上皮完整性、紧密连接蛋白和粘附分子的作用。方法:本研究以断奶前BL/6幼犬(3周龄,同窝)为模型动物。我们给实验幼崽(n=7)提供未经消毒的劣质水,以产生非特异性腹泻症状。然后连续3周在腹泻幼崽中添加嗜酸乳杆菌(LA)。对照组(n=5)给予无菌水,不给予LA。分别于第0天、第7天、第14天和第21天进行取样和分析。采用qRT-PCR检测肠道黏膜促炎因子(IL-6、TNF-a)和紧密连接蛋白(TJPs)的表达。采用夹心ELISA法检测血清细胞因子水平。结果:实时qRT-PCR结果显示,肠道细胞骨架完整性被破坏,ZO-1、Occludin、Claudin-1、Claudin-5和JAM mRNA表达降低。然而,Claudin-4没有受到影响,并且与对照幼崽一样具有更高的表达。有趣的是,补充嗜酸乳杆菌被发现可以维持肠道完整性并有效减轻腹泻症状。与对照组幼鼠一样,嗜酸乳杆菌组幼鼠表现出更高的肠道上皮TJPs和粘附分子的表达。此外,与对照组相比,患病幼崽血清中IL-6和TNF-α的含量显著增加。结论:我们认为补充嗜酸乳杆菌可能会协调肠道健康和对非特异性腹泻的免疫平衡。
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