Promising protective treatment potential of endophytic bacterium Rhizobium aegyptiacum for ulcerative colitis in rats.

IF 4.7 3区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY
Engy Elekhnawy, Duaa Eliwa, Sebaey Mahgoub, Sameh Magdeldin, Ehssan Moglad, Sarah Ibrahim, Asmaa Ramadan Azzam, Rehab Ahmed, Walaa A Negm
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引用次数: 0

Abstract

Ulcerative colitis (UC) is an inflammatory condition of the intestine, resulting from an increase in oxidative stress and pro-inflammatory mediators. In this study, the extract of endophytic bacterium Rhizobium aegyptiacum was prepared for the first time using liquid chromatography-mass spectrometry (LC-MS). In addition, also for the first time, the protective potential of R. aegyptiacum was revealed using an in vivo rat model of UC. The animals were grouped into four categories: normal control (group I), R. aegyptiacum (group II), acetic acid (AA)‍-induced UC (group III), and R. aegyptiacum-treated AA-induced UC (group IV). In group IV, R. aegyptiacum was administered at 0.2 mg/kg daily for one week before and two weeks after the induction of UC. After sacrificing the rats on the last day of the experiment, colon tissues were collected and subjected to histological, immunohistochemical, and biochemical investigations. There was a remarkable improvement in the histological findings of the colon tissues in group IV, as revealed by hematoxylin and eosin (H&E) staining, Masson's trichrome staining, and periodic acid-Schiff (PAS) staining. Normal mucosal surfaces covered with a straight, intact, and thin brush border were revealed. Goblet cells appeared magenta in color, and there was a significant decrease in the distribution of collagen fibers in the mucosa and submucosal connective tissues. All these findings were comparable to the respective characteristics of the control group. Regarding cyclooxygenase-2 (COX-2) immunostaining, a weak immune reaction was shown in most cells. Moreover, the colon tissues were examined using a scanning electron microscope, which confirmed the results of histological assessment. A regular polygonal unit pattern was seen with crypt orifices of different sizes and numerous goblet cells. Furthermore, the levels of catalase (CAT), myeloperoxidase (MPO), nitric oxide (NO), interleukin-6 (IL-6), and interlukin-1β (IL-1β) were determined in the colonic tissues of the different groups using colorimetric assay and enzyme-linked immunosorbent assay (ELISA). In comparison with group III, group IV exhibited a significant rise (P<0.05) in the CAT level but a substantial decline (P<0.05) in the NO, MPO, and inflammatory cytokine (IL-6 and IL-1β) levels. Based on reverse transcription-quantitative polymerase chain reaction (RT-qPCR), the tumor necrosis factor-‍α (TNF-‍α) gene expression was upregulated in group III, which was significantly downregulated (P<0.05) by treatment with R. aegyptiacum in group IV. On the contrary, the heme oxygenase-1 (HO-1) gene was substantially upregulated in group IV. Our findings imply that the oral consumption of R. aegyptiacum ameliorates AA-induced UC in rats by restoring and reestablishing the mucosal integrity, in addition to its anti-oxidant and anti-inflammatory effects. Accordingly, R. aegyptiacum is potentially effective and beneficial in human UC therapy, which needs to be further investigated in future work.

埃及根瘤根瘤菌对大鼠溃疡性结肠炎的保护性治疗潜力。
溃疡性结肠炎(UC)是一种肠道炎症,由氧化应激和促炎介质的增加引起。本研究首次采用液相色谱-质谱联用技术对内生细菌埃及根瘤菌的提取物进行了制备。此外,也首次通过UC大鼠体内模型揭示了埃及褐霉的保护潜力。将实验动物分为正常对照组(ⅰ组)、埃及伊蚊(ⅱ组)、醋酸(AA)‍诱导的UC(ⅲ组)和埃及伊蚊经AA处理的UC(ⅳ组)。ⅳ组在诱导UC前1周和诱导UC后2周,每天给药埃及伊蚊0.2 mg/kg。实验最后一天处死大鼠,收集结肠组织进行组织学、免疫组化和生化检查。苏木精伊红(H&E)染色、马松三色染色和周期性酸-希夫(PAS)染色显示,IV组结肠组织组织学表现明显改善。正常粘膜表面覆盖着直的、完整的、薄的刷状边缘。杯状细胞呈洋红色,粘膜及粘膜下结缔组织胶原纤维分布明显减少。所有这些发现都与对照组的各自特征相当。环氧化酶-2 (COX-2)免疫染色在大多数细胞中显示弱免疫反应。并对结肠组织进行扫描电镜检查,证实了组织学评价结果。可见规则的多边形单位模式,有不同大小的隐窝孔和大量的杯状细胞。采用比色法和酶联免疫吸附法(ELISA)检测各组大鼠结肠组织过氧化氢酶(CAT)、髓过氧化物酶(MPO)、一氧化氮(NO)、白细胞介素-6 (IL-6)和白细胞介素-1β (IL-1β)水平。与III组相比,IV组显著升高(PPTNF-‍α)基因表达上调,而III组显著下调(埃及红霉在IV组)。相反,血红素加氧酶-1 (HO-1)基因在IV组显著上调。我们的研究结果表明,口服埃及红霉通过恢复和重建粘膜完整性来改善aa诱导的大鼠UC。除了它的抗氧化和抗炎作用。因此,埃及雷公杆菌在人类UC治疗中具有潜在的有效和有益作用,需要在今后的工作中进一步研究。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Journal of Zhejiang University SCIENCE B
Journal of Zhejiang University SCIENCE B 生物-生化与分子生物学
CiteScore
8.70
自引率
13.70%
发文量
2125
审稿时长
3.0 months
期刊介绍: Journal of Zheijang University SCIENCE B - Biomedicine & Biotechnology is an international journal that aims to present the latest development and achievements in scientific research in China and abroad to the world’s scientific community. JZUS-B covers research in Biomedicine and Biotechnology and Biochemistry and topics related to life science subjects, such as Plant and Animal Sciences, Environment and Resource etc.
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