Effect of Manuka honey on IL-8 secretion in HDFa and the role of transcription factor activator protein (Ap-1) in inflammatory response
Cardiff Metropolitan University
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Wound healing is a biological process function to aid in the reduction of blood loss in the incidence of an injury. It involves four complex phases and they are: haemostasis, inflammation, proliferation and ends with remodelling. The process involves the activation of multiple cell lines including, fibroblasts, keratinocytes, endothelial, macrophages, and platelets cells. Besides that, it is regulated by many cytokines and growth factors such as fibroblast growth factor (FGF), and tumour necrosis factor-alpha (TNF- α). In diabetic people, healing of wounds is impaired or not fully healed which leads to further complications such as infection and trauma. Also, they are prone to develop non-healing diabetic foot ulcers (DFUs) Wound ulcers is a major issue for people with diabetes and thus causes financial difficulties to the national health service (NHS). However, it has been proposed that Manuka honey can aid in the increase of pro-inflammatory and anti-inflammatory cytokines during inflammation in the process of wound healing. The aim of this study is to investigate into the effect of Manuka honey on the release of IL-8 from fibroblast cells and investigating a possible inflammatory pathway mechanism by using AP1 inhibitor. The hypothesis therefore, is that the activator protein 1 (AP1) is responsible in activating the mechanism pathway to release interleukin-8 (IL-8) in inflammation. The experiment was conducted by investigating pro-inflammatory cytokines of human (TNF α) and (IL-1β) in human dermal fibroblast cells (HDFa) to initiate the release of IL-8. This follows by adding 1.0% of Manuka honey and then 1um and 10um of (Ap-1) inhibitor that was introduced to the cells. It was then analysed by enzyme-linked immunosorbent assay (ELISA). Results obtained showed that IL-1β and TNFα has shown a highly significant impact on the release of IL-8 when compared to the control. IL-1β showing a p-value of (0.0013) and TNFα showing a p-value of (0.0284). Manuka honey increases the release of IL-8 with a significant difference to the control (p-value: 0.0009). when using 10uM of Ap-1 inhibitor it significantly inhibits the concentration of IL-8 release in the presence of IL-1 β (p-value:0.0292), but not significantly when TNFα is present. However, 1uM of Ap-1 inhibitor it significantly raises concentration of IL-8 in the presence of TNFα (p-value: 0.0154). Moreover, using 10uM of Ap-1 inhibitor had significantly suppressed the release of IL-8 when used with Manuka honey, giving a p-value of (p-value: <0.0001). To conclude, high concentration of Ap-1 inhibitor decrease the IL-8 release if HDFa in the presence of IL-1 β and Manuka honey. This suggests that Ap-1 initiated the transcription mechanism of IL-8 and Manuka honey does play a role in inflammation. However, further research has to be carried to improve findings.
BSc (Hons) Biomedical Science
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