Full Text:   <824>

Summary:  <41>

Suppl. Mater.: 

CLC number: 

On-line Access: 2025-03-13

Received: 2023-10-27

Revision Accepted: 2024-02-20

Crosschecked: 2025-03-13

Cited: 0

Clicked: 1246

Citations:  Bibtex RefMan EndNote GB/T7714

 ORCID:

Engy ELEKHNAWY

https://orcid.org/0000-0001-8287-1026

Walaa A. NEGM

https://orcid.org/0000-0003-0463-8047

-   Go to

Article info.
Open peer comments

Journal of Zhejiang University SCIENCE B 2025 Vol.26 No.3 P.286-301

http://doi.org/10.1631/jzus.B2300777


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


Author(s):  Engy ELEKHNAWY, Duaa ELIWA, Sebaey MAHGOUB, Sameh MAGDELDIN, Ehssan MOGLAD, Sarah IBRAHIM, Asmaa Ramadan AZZAM, Rehab AHMED, Walaa A. NEGM

Affiliation(s):  Pharmaceutical Microbiology Department, Faculty of Pharmacy, Tanta University, Tanta31527,Egypt; more

Corresponding email(s):   engy.ali@pharm.tanta.edu.eg, walaa.negm@pharm.tanta.edu.eg

Key Words:  Inflammatory bowel syndrome, Liquid chromatography-tandem mass spectrometry (LC-MS/MS), Scanning electron microscopy (SEM), Histology, Immunohistochemistry, Reverse transcription-quantitative polymerase chain reaction (RT-qPCR)


Engy ELEKHNAWY, Duaa ELIWA, Sebaey MAHGOUB, Sameh MAGDELDIN, Ehssan MOGLAD, Sarah IBRAHIM, Asmaa Ramadan AZZAM, Rehab AHMED, Walaa A. NEGM. Promising protective treatment potential of endophytic bacterium Rhizobium aegyptiacum for ulcerative colitis in rats[J]. Journal of Zhejiang University Science B, 2025, 26(3): 286-301.

@article{title="Promising protective treatment potential of endophytic bacterium Rhizobium aegyptiacum for ulcerative colitis in rats",
author="Engy ELEKHNAWY, Duaa ELIWA, Sebaey MAHGOUB, Sameh MAGDELDIN, Ehssan MOGLAD, Sarah IBRAHIM, Asmaa Ramadan AZZAM, Rehab AHMED, Walaa A. NEGM",
journal="Journal of Zhejiang University Science B",
volume="26",
number="3",
pages="286-301",
year="2025",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B2300777"
}

%0 Journal Article
%T Promising protective treatment potential of endophytic bacterium Rhizobium aegyptiacum for ulcerative colitis in rats
%A Engy ELEKHNAWY
%A Duaa ELIWA
%A Sebaey MAHGOUB
%A Sameh MAGDELDIN
%A Ehssan MOGLAD
%A Sarah IBRAHIM
%A Asmaa Ramadan AZZAM
%A Rehab AHMED
%A Walaa A. NEGM
%J Journal of Zhejiang University SCIENCE B
%V 26
%N 3
%P 286-301
%@ 1673-1581
%D 2025
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B2300777

TY - JOUR
T1 - Promising protective treatment potential of endophytic bacterium Rhizobium aegyptiacum for ulcerative colitis in rats
A1 - Engy ELEKHNAWY
A1 - Duaa ELIWA
A1 - Sebaey MAHGOUB
A1 - Sameh MAGDELDIN
A1 - Ehssan MOGLAD
A1 - Sarah IBRAHIM
A1 - Asmaa Ramadan AZZAM
A1 - Rehab AHMED
A1 - Walaa A. NEGM
J0 - Journal of Zhejiang University Science B
VL - 26
IS - 3
SP - 286
EP - 301
%@ 1673-1581
Y1 - 2025
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B2300777


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 bacteriumRhizobium aegyptiacum was prepared for the first time using liquid chromatography-mass spectrometry (LC-MS). In addition, also for the first time, the protective potential ofR. 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 withR. 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 ofR. 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.

内生菌Rhizobium aegyptiacum对大鼠溃疡性结肠炎的潜在保护性治疗效果研究

Engy ELEKHNAWY1,Duaa ELIWA2,Sebaey MAHGOUB3,Sameh MAGDELDIN3,4,Ehssan MOGLAD5,Sarah IBRAHIM6,Asmaa Ramadan AZZAM6,Rehab AHMED7,Walaa A. NEGM2
1Pharmaceutical Microbiology Department, Faculty of Pharmacy, Tanta University, Tanta 31527, Egypt
2Department of Pharmacognosy, Faculty of Pharmacy, Tanta University, Tanta 31527, Egypt
3Proteomics and Metabolomics Research Program, Department of Basic Research, Children’s Cancer Hospital 57357, Cairo 11441, Egypt
4Department of Physiology, Faculty of Veterinary Medicine, Suez Canal University, Ismailia 41522, Egypt
5Department of Pharmaceutics, College of Pharmacy, Prince Sattam bin Abdulaziz University, P.O. Box 173, Alkharj 11942, Saudi Arabia
6Human Anatomy and Embryology Department, Faculty of Medicine, Tanta University, Tanta 31527, Egypt
7Division of Microbiology, Immunology and Biotechnology, Department of Natural Products and Alternative Medicine, Faculty of Pharmacy, University of Tabuk, Tabuk 47713, Saudi Arabia
摘要:溃疡性结肠炎(UC)是由氧化应激和炎性介质增加所导致的一种非特异性的肠道炎症性疾病。本研究首次通过液相色谱-质谱联用技术制备了植物内生细菌Rhizobium aegyptiacum提取物,并通过大鼠模型验证其在体内对UC的保护潜力。实验共分为四组,分别为正常对照组(组I)、R. aegyptiacum组(组II)、经醋酸(AA)诱导的UC组(组III)和经R. aegyptiacum治疗的AA诱导UC组(组IV)。其中,组IV大鼠在UC诱导前一周和后两周每天给予R. aegyptiacum处理(0.2 mg/kg)。在实验的最后一天,我们收集各组大鼠结肠组织进行组织学、免疫组化和生化学检测。苏木精-伊红染色、马松染色和PAS染色结果显示,组IV大鼠的结肠出现明显的组织学改善。与对照组相似,其粘膜表面正常,覆盖着平直、完整且薄的刷状边缘;杯状细胞呈品红色,粘膜和黏膜下结缔组织中的胶原纤维分布显著减少。环氧合酶-2(COX-2)免疫染色显示大多数细胞出现弱免疫反应。结肠组织的扫描电镜检查结果与上述组织学评估一致,不同大小的隐窝孔和大量的杯状细胞显示出规律的多边形单元模式。不同组中大鼠结肠组织过氧化氢酶(CAT)、髓过氧化物酶(MPO)、一氧化氮(NO)、白介素-6(IL-6)和白介素-1β(IL-1β)水平的测定结果显示,与组III相比,组IV的CAT水平显著上升(P<0.05),但NO、MPO和炎症细胞因子(IL-6和IL-1β)水平显著降低(P<0.05)。逆转录定量聚合酶链反应(RT-qPCR)结果显示,经AA诱导的UC组(组III)中TNF-α基因表达上调,而在经R. aegyptiacum治疗的AA诱导UC组(组IV)中则呈显著下调(P<0.05);与此相反,组IV中的HO-1基因则显著上调。本研究实验结果表明,口服R. aegyptiacum能恢复和重建粘膜完整,进而改善AA诱导的大鼠UC,并具有抗氧化和抗炎作用。综上,R. aegyptiacum在人类UC治疗中可能具有潜在价值,但仍需进一步验证。

关键词:炎症性肠综合征;液相色谱-串联质谱(LC-MS/MS);扫描电子显微镜(SEM);组织学;免疫组织化学;逆转录定量聚合酶链反应(RT-qPCR)

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

Reference

[1]AbdallahHMI,AmmarNM,AbdelhameedMF,et al.,2020.Protective mechanism ofAcacia saligna butanol extract and its nano-formulations against ulcerative colitis in rats as revealed via biochemical and metabolomic assays.Biology (Basel),9(8):195.

[2]AbdelazizA,SonbolF,ElbannaT,et al.,2019.Exposure to sublethal concentrations of benzalkonium chloride induces antimicrobial resistance and cellular changes inKlebsiellae pneumoniae clinical isolates.Microb Drug Resist,25(5):631-638.

[3]AdaminaM,FeakinsR,IacucciM,et al.,2021.ECCO topical review optimising reporting in surgery, endoscopy, and histopathology: collaboration between S-ECCO, EduCom, H-ECCO.J Crohns Colitis,15(7):1089-1105.

[4]AlherzFA,NegmWA,ElekhnawyE,et al.,2022.Silver nanoparticles prepared usingEncephalartos laurentianus De Wild leaf extract have inhibitory activity againstCandida albicans clinical isolates.J Fungi (Basel),8(10):1005.

[5]AlmukainziM,El-MasryTA,NegmWA,et al.,2022.Co-delivery of gentiopicroside and thymoquinone using electrospun m-PEG/PVP nanofibers: in-vitro and in vivo studies for antibacterial wound dressing in diabetic rats.Int J Pharm,625:122106.

[6]AlotaibiB,El-MasryTA,ElekhnawyE,et al.,2022.Aqueous core epigallocatechin gallate PLGA nanocapsules: characterization, antibacterial activity against uropathogens, andin vivo reno-protective effect in cisplatin induced nephrotoxicity.Drug Deliv,29(1):1848-1862.

[7]AnsariMN,RehmanNU,KarimA,et al.,2021.Role of oxidative stress and inflammatory cytokines (TNF-α and IL-6) in acetic acid-induced ulcerative colitis in rats: ameliorated byOtostegia fruticosa.Life (Basel),11(3):195.

[8]AttallahNGM,Al-FakhranyOM,ElekhnawyE,et al.,2022a.Anti-biofilm and antibacterial activities ofCycas media R.Br secondary metabolites:in silico, in vitro, and in vivo approaches. Antibiotics (Basel),11(8):993.

[9]AttallahNGM,El-SherbeniSA,El-KademAH,et al.,2022b.Elucidation of the metabolite profile ofYucca gigantea and assessment of its cytotoxic, antimicrobial, and anti-inflammatory activities.Molecules,27(4):1329.

[10]AzizN,KimMY,ChoJY,2018.Anti-inflammatory effects of luteolin: a review of in vitro, in vivo, and in silico studies.J Ethnopharmacol,225:342-358.

[11]BancroftJD,GambleM,2008.Theory and Practice of Histological Techniques, 6th Ed.Churchill Livingstone,Philadelphia, USA.

[12]Ben-HorinS,NovackL,MaoR,et al.,2022.Efficacy of biologic drugs in short-duration versus long-duration inflammatory bowel disease: a systematic review and an individual-patient data meta-analysis of randomized controlled trials.Gastroenterology,162(2):482-494.

[13]BertevelloPL,LogulloÂF,NonogakiS,et al.,2005.Immunohistochemical assessment of mucosal cytokine profile in acetic acid experimental colitis.Clinics (Sao Paulo),60(4):277-286.

[14]BlacherE,LevyM,TatirovskyE,et al.,2017.Microbiome-modulated metabolites at the interface of host immunity.J Immunol,198(2):572-580.

[15]BruscoliS,FeboM,RiccardiC,et al.,2021.Glucocorticoid therapy in inflammatory bowel disease: mechanisms and clinical practice.Front Immunol,12:691480.

[16]ChecaJ,AranJM,2020.Reactive oxygen species: drivers of physiological and pathological processes.J Inflamm Res,13:1057-1073.

[17]DignassA,EliakimR,MagroF,et al.,2012.Second European evidence-based consensus on the diagnosis and management of ulcerative colitis part 1: definitions and diagnosis.J Crohns Colitis,6(10):965-990.

[18]ElekhnawyE,NegmWA,2022.The potential application of probiotics for the prevention and treatment of COVID-19.Egypt J Med Hum Genet,23:36.

[19]ElmongyEI,NegmWA,ElekhnawyE,et al.,2022.Antidiarrheal and antibacterial activities of Monterey cypress phytochemicals: in vivo and in vitro approach.Molecules,27(2):346.

[20]FanMM,XiangG,ChenJW,et al.,2020.Libertellenone M, a diterpene derived from an endophytic fungus Phomopsis sp. S12,protects against DSS-induced colitis via inhibiting both nuclear translocation of NF-κB and NLRP3 inflammasome activation. Int Immunopharmacol,80:106144.

[21]GautamMK,GoelS,GhatuleRR,et al.,2013.Curative effect ofTerminalia chebula extract on acetic acid-induced experimental colitis: role of antioxidants, free radicals and acute inflammatory marker.Inflammopharmacology,21(5):377-383.

[22]HsiaoYP,ChenHL,TsaiJN,et al.,2021.Administration ofLactobacillus reuteri combined withClostridium butyricum attenuates cisplatin-induced renal damage by gut microbiota reconstitution, increasing butyric acid production, and suppressing renal inflammation.Nutrients,13(8):2792.

[23]KaurL,GordonM,BainesPA,et al.,2020.Probiotics for induction of remission in ulcerative colitis.Cochrane Database Syst Rev,3(3):CD005573.

[24]KhairyH,SalehH,BadrAM,et al.,2018.Therapeutic efficacy of osthole against dinitrobenzene sulphonic acid induced-colitis in rats.Biomed Pharmacother,100:42-51.

[25]KhareT,PalakurthiSS,ShahBM,et al.,2020.Natural product-based nanomedicine in treatment of inflammatory bowel disease.Int J Mol Sci,21(11):3956.

[26]LaneDJ,1991.16S/23S rRNA sequencing. In: Stackebrandt E, Goodfellow M (Eds.),Nucleic Acid Techniques in Bacterial Systematics.John Wiley & Sons,Chichester, p.115-147.

[27]LavelleA,SokolH,2020.Gut microbiota-derived metabolites as key actors in inflammatory bowel disease.Nat Rev Gastroenterol Hepatol,17(4):223-237.

[28]le LouppAG,Bach-NgohouK,BourreilleA,et al.,2015.Activation of the prostaglandin D2 metabolic pathway in Crohn’s disease: involvement of the enteric nervous system.BMC Gastroenterol,15:112.

[29]LiX,LiuX,ZhangYF,et al.,2020.Protective effect of Gloeostereum incarnatum on ulcerative colitis via modulation of Nrf2/NF-κB signaling in C57BL/6 mice.Mol Med Rep,22(4):3418-3428.

[30]LiuXL,DongMS,ChenXH,et al.,2007.Antioxidant activity and phenolics of an endophyticXylaria sp.from Ginkgo biloba. Food Chem,105(2):548-554.

[31]LiuY,WeiWH,LiangSW,et al.,2022.Esculentoside A could attenuate apoptosis and inflammation in TNBS-induced ulcerative colitis via inhibiting the nuclear translocation of NF-κB.Ann Transl Med,10(14):771.

[32]LivakKJ,SchmittgenTD,2001.Analysis of relative gene expression data using real-time quantitative PCR and the2-ΔΔCTmethod.Methods,25(4):402-408.

[33]MiH,LiuFB,LiHW,et al.,2017.Anti-inflammatory effect of Chang-An-Shuan on TNBS-induced experimental colitis in rats.BMC Complement Altern Med,17:315.

[34]PeyrottesA,CoquantG,BrotL,et al.,2020.Anti-inflammatory effects of analogues ofN-acyl homoserine lactones on eukaryotic cells.Int J Mol Sci,21(24):9448.

[35]Piechota-PolanczykA,FichnaJ,2014.Review article: the role of oxidative stress in pathogenesis and treatment of inflammatory bowel diseases.Naunyn Schmiedebergs Arch Pharmacol,387(7):605-620.

[36]PopovJ,CaputiV,NandeeshaN,et al.,2021.Microbiota-immune interactions in ulcerative colitis and colitis associated cancer and emerging microbiota-based therapies.Int J Mol Sci,22(21):11365.

[37]Quettier-DeleuC,GressierB,VasseurJ,et al.,2000.Phenolic compounds and antioxidant activities of buckwheat (Fagopyrum esculentum Moench) hulls and flour.J Ethnopharmacol,72(1-2):35-42.

[38]RaishM,ShahidM,Bin JardanYA,et al.,2021.Gastroprotective effect of sinapic acid on ethanol-induced gastric ulcers in rats: involvement of Nrf2/HO-1 and NF-κB signaling and antiapoptotic role.Front Pharmacol,12:622815.

[39]RanZH,ChenC,XiaoSD,2008.Epigallocatechin-3-gallate ameliorates rats colitis induced by acetic acid.Biomed Pharmacother,62(3):189-196.

[40]RanaKL,KourD,KaurT,et al.,2020.Endophytic microbes: biodiversity, plant growth-promoting mechanisms and potential applications for agricultural sustainability.Antonie van Leeuwenhoek,113(8):1075-1107.

[41]ShahidM,RaishM,AhmadA,et al.,2022.Sinapic acid ameli

[42]orates acetic acid-induced ulcerative colitis in rats by suppressing inflammation,stressoxidative, andapoptosis.Molecules,27(13):4139.

[43]SinghM,KumarA,SinghR,et al.,2017.Endophytic bacteria: a new source of bioactive compounds.3Biotech,7(5):315.

[44]StrobelG,DaisyB,2003.Bioprospecting for microbial endophytes and their natural products.Microbiol Mol Biol Rev,67(4):491-502.

[45]TwaijBM,HasanMN,2022.Bioactive secondary metabolites from plant sources: types, synthesis, and their therapeutic uses.Int J Plant Biol,13(1):4-14.

[46]VavrickaSR,BrunL,BallabeniP,et al.,2011.Frequency and risk factors for extraintestinal manifestations in the Swiss inflammatory bowel disease cohort.Am J Gastroenterol,106(1):110-119.

[47]WangD,DuBoisRN,2010.The role of COX-2 in intestinal inflammation and colorectal cancer.Oncogene,29(6):781-788.

[48]WangQH,ZhiTX,HanPP,et al.,2021.Potential anti-inflammatory activity of walnut protein derived peptide leucine-proline-phenylalanine in lipopolysaccharides-irritated RAW264.7 cells.Food Agric Immunol,32(1):663-678.

[49]XueJC,YuanS,HouXT,et al.,2023.Natural products modulate NLRP3 in ulcerative colitis.Front Pharmacol,14:1265825.

Open peer comments: Debate/Discuss/Question/Opinion

<1>

Please provide your name, email address and a comment





Journal of Zhejiang University-SCIENCE, 38 Zheda Road, Hangzhou 310027, China
Tel: +86-571-87952783; E-mail: cjzhang@zju.edu.cn
Copyright © 2000 - 2025 Journal of Zhejiang University-SCIENCE