Evaluation of honey extract activity against the expression level in IL-1β of Helicobacter pylori isolates from suspected gastric ulcer patients | ||||
| Microbes and Infectious Diseases | ||||
| Articles in Press, Accepted Manuscript, Available Online from 02 September 2025 | ||||
| Document Type: Original Article | ||||
| DOI: 10.21608/mid.2025.410670.3081 | ||||
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| Authors | ||||
Iman Fadhil Abdul-Husin1; Tsahel H. Al-Dulaimi  2; Farah Mohammed Saeed 2; Hasanain Khaleel Shareef 3, 4; Israa Adnan Ibraheam 2; Raghda A. Razaq 2; Rana M.K. AL.bayati 2
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| 1Applied biotechnology department, Collage of biotechnology, Al-Qasim Green University, Babylon, Iraq | ||||
| 2Department of Biology, College of Science for Women, University of Babylon, Babylon, Iraq | ||||
| 3Al-Mustaqbal University, College of Science , Department of Medical Biotechnology, 51001, Babylon, Iraq. | ||||
| 4University of Babylon, College of Science for Women, Biology Department, Babylon, Iraq | ||||
| Abstract | ||||
| Background: Among the strong health contenders is the spiral-shaped bacterium Helicobacter pylori. In addition to its inhibitor effect on the secretion function in epithelial cells for the period of H. pylori infection; the IL-1β plays a role in initiating and exacerbating the inflammatory response and is crucial for eliminating ingested bacteria, which often express cytotoxic cytosolic gene A (cagA) strains of Helicobacter pylori. The antibacterial effect of antibiotics is not entirely effective in such an infected area. Therefore, natural compounds such as honey have been used in current studies to investigate their antibacterial activity against Helicobacter pylori infection. Aims: Evaluation of honey extract activity against cytotoxic associated gene A (cagA) strains of Helicobacter pylori was isolated from biopsies of patients with suspected gastric infection, and IL-1β levels assessed as an immune response measure to lyse and clear the killed bacteria. Methods: Only 8 biopsies were positive for H. pylori infection obtained from the 63 males and 37 females patients at different age groups in which H. pylori infection was presumed in the progression study. They were expected diagnostic for ‘upper’ gastrointestinal endoscopy at Endoscopy part of AL- Hilla Teaching Hospital in Babylone“ through the dated December 2024 towards Febuary 2025.Each biopsy specimens was transport in Stuart medium, then suspension with a normal saline and after that they were streaked on the Skirrow Colombia blood agar, incubated for 2-7 days at 37 ºC, .The optimistic growth was identified reliant on morphological and microscopically features and biochemical tests. IL-1β protein concentration measurement was done according to the procedure described by Yamaoka ,et al (1999) in the biopsy specimens which was also used later for quantitative culture of H. pylori. Honey extract purification was done by inoculating onto blood agar and incubating aerobically at 37°C for 48 hours. To make 1.5 ml micro centrifuge tubes with a final concentration of 10 mg/ml of honey extract, the tubes were filled with distilled water and mixed to a stock solution. Concentrations ranging from 10 mg/ml to 0.01 mg/ml were achieved by preparing two-fold serial dilutions of the stock solution in a 96-well plate using Muller-Hinton broth. After the end of the known incubation period, cDNA was already amplified by quantitative RT-PCR (qPCR) using forward and reverse primers specific for the applicable target gene, as well as a reference gene for normalization. An initial activation, denaturation, and annealing phase were included of the qPCR program. The ΔΔCT method was used to compute the fold change by comparing the target gene's cycle threshold (CT) to the reference gene's CT. Gene expression investigations can benefit from this simplified protocol's RNA extraction, cDNA synthesis, and qPCR analysis capabilities. Results: All the isolates of H. Pylori were small White Colonies, positive to for both catalase, oxidase and urease tests. The results of MIC value the honey extract against tested H. Pylori were ranged at ( 50 mg/ml-0.09 mg/ml). The concentration 0.7 mg/ml was inhibiting the growth of isolates (4/8) 50%, the concentration 1.56 mg/ml was inhibiting the growth of (3/8) 37.5 %, and concentration 0.3 mg/ml (1/8)12.5%. Results indicated that expression levels of Cag A gene was highly being decrease significantly in (after) group with a fold change of (0.239) compared to expression levels of current gene with a fold change of (1.686) before the bacterial treatment with honey extract. IL-1β concentration was significantly higher in the all biopsy of H. pylori-infected patients (p ≤ 0.05) at age group (39-49) than in the uninfected individual enrolled in current study while there are no significant difference concerning sex parameter and the higher concentration of IL-1β. Conclusion: H. pylori virulence genes ( Cag A) are highly prevalent among patients with Gastric ulcer symptom .The Cag A genes are significantly associated with the clinical outcomes especially chronic active gastritis. Expression levels of Cag A gene was highly being decrease significantly after treatment with honey extract in H. pylori isolated. | ||||
| Keywords | ||||
| Honey Extract; IL-1β; Helicobacter pylori; Gastric ulcer patients | ||||
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