Effect of ethanolic propolis extract on TNF-α, caspase-3 expression and histopathological findings in hypoxic skin flaps: an experimental study in rats

Kevin Piether; Amru Sungkar; Kristanto Y. Yarsa; Brian Wasita

doi:10.18203/2349-2902.isj20262447

Authors

Kevin Piether Department of General Surgery, Sebelas Maret University, Surakarta, Central of Java, Indonesia
Amru Sungkar Department of Plastic Surgery, Sebelas Maret University, Surakarta, Central of Java, Indonesia https://orcid.org/0009-0009-7963-0745
Kristanto Y. Yarsa Department of Oncology Surgery, Sebelas Maret University, Surakarta, Central of Java, Indonesia
Brian Wasita Department of Anatomic Pathology, Sebelas Maret University, Surakarta, Central of Java, Indonesia

DOI:

https://doi.org/10.18203/2349-2902.isj20262447

Keywords:

Propolis, Skin flap, Hypoxia, TNF-α, Caspase-3, Apoptosis

Abstract

Background: Hypoxia remains a major cause of skin flap failure due to excessive production of reactive oxygen species (ROS), which triggers inflammation and apoptosis through increased expression of tumor necrosis factor-alpha (TNF-α) and caspase-3. Propolis contains bioactive compounds with antioxidant and anti-inflammatory properties that may improve skin flap survival under hypoxic conditions. This study aimed to evaluate the effect of ethanolic propolis extract on TNF-α expression, caspase-3 expression, and histopathological changes in a hypoxic skin flap model.

Methods: An experimental post-test only control group study was conducted using 24 male Rattus norvegicus divided into four groups (n=6/group): control, P1, P2, and P3. Different flap dimensions (1/3, 1/2, and 2/3) were used to induce varying degrees of hypoxia. Ethanolic extract of Gunung Lawu propolis was administered orally at a dose of 800 mg/kg bw/day for seven days. TNF-α and caspase-3 levels were measured using enzyme-linked immunosorbent assay (ELISA). Histopathological evaluation included necrosis, acute inflammation, and re-epithelialization. Statistical analysis was performed using one-way ANOVA followed by post hoc tests and Kruskal–Wallis analysis.

Results: Propolis administration significantly reduced TNF-α levels in all treatment groups compared with controls (p<0.001), with the greatest reduction observed in the P3 group (55.5%). Caspase-3 expression was also significantly decreased (p<0.001), with the highest reduction found in the P3 group (79.5%). Histopathological assessment demonstrated lower necrosis and acute inflammation scores and higher re-epithelialization scores in treatment groups; however, these differences were not statistically significant (p>0.05).

Conclusions: Ethanolic propolis extract effectively attenuated inflammatory and apoptotic responses in hypoxic skin flaps by reducing TNF-α and caspase-3 expression. Although histopathological improvements were observed, they did not reach statistical significance. Propolis may serve as a promising adjunctive therapy to enhance skin flap viability.

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