Effects of Philippine Stingless Bee (Tetragonula biroi FRIESE) Propolis on Gene Expression pf TH1-Related Cytokines in Salmonella Typhimurium-Stimulated Philippines Banaba Native Chicken Embryonic Fibroblasts

Gerald Z. Chu

Date

2024

Degree

Doctor of Veterinary Medicine

Adviser/Committee Chair

Mark Joseph M. Desamero

Co-adviser

Cherry P. Fernandez-Colorado

Abstract

Background: Salmonellosis is the most common food-borne disease in humans worldwide, with Salmonella Typhimurium (S. Typhimurium) identified to be the main causative agent. The increase in antibiotic resistance against poultry pathogens causes significant economic losses and devastating public health concerns. Despite the increasing interest in the therapeutic properties of stingless bee propolis, the biological activity of the Philippine stingless propolis remains poorly understood and sparingly investigated. Herein, we have investigated the effect of ethanolic extract of Philippine stingless propolis on the expression of Th-1 related cytokines in S. Typhimurium stimulated chicken embryonic fibroblasts (CEFs). Methods: CEFs, isolated from 9-day-old embryos, were assigned into four experimental groups namely: PBS-treated mock-stimulated group (NC), S. Typhimurium-stimulated control group (ST), EEP-treated group (EEP), and S. Typhimurium-stimulated and EEP-treated group (ST+EEP). The highest concentration of EEP that did not produce more than 20% growth inhibition of CEFs was initially determined using trypan blue exclusion assay. After which, cells were incubated with different treatments for 24 hours and then subjected to quantitative reverse-transcription (qRT) PCR to determine the gene expression levels of Thl-related cytokines. Results: Cell viability assay showed that 100 g/ml corresponded to the optimum concentration of EEP that preserve the viability as well as the cellular integrity of CEFs. qRT PCR analysis of Thl-related cytokines unveiled that mRNA transcript levels of IN-y were significantly upregulated in the ST+EEP group whereas the expression levels of IL-2 and IL-8 were consistently and highly significantly increased in the ST alone group as compared to the corresponding NC control group. In addition, gene expression levels of IFN-y were observed to be significantly upregulated in the ST+EEP group with respect to the ST alone group. Conversely, the expression levels of both IL-2 and IL-8 were considerably downregulated in the former group as opposed to the latter group. Finally, as contrasted to the EEP alone group, the gene expression levels of both IL-2 and IL-8 were significantly increased in the ST alone group. Conclusion: Altogether, the results of this study suggest that the Philippine stingless bee propolis specifically modulates the gene transcription of Thl-related cytokines in response to S. Typhimurium infection in vitro.

Language

English

Location

UPLB College of Veterinary Medicine (CVM)

Notes

Awards : Teodulo C. Topacio Sr. Best Undergraduate Thesis Award in Veterinary Microbiology

Document Type

Thesis

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