Background: Extensive wound closure due to trauma, infection and post tumor excision often requires a skin graft procedure. One of the successful factors of skin graft is the good quality of recipient tissue vascularization. Reactive oxygen species (ROS) is a factor that plays a role in vascularization, an excessive accumulation of ROS can interfere with the process of vascularization. Administration of the antioxidant can keep the ROS levels from being excessive thus producing the recipient vascularization for a successful skin graft. Propolis has an effect of antioxidant, inflammation and immunomodulator, anti-ROS and anti-inflammatory effects due to the large polyphenol content. This study aims to examine the antioxidant effect of propolis ethanol extract Lawu mountain (EEP) in a rats skin graft model by examining its effect on oxidative stress levels (Malondialdehyde blood levels/MDA), effect on inflammation (IL-6 blood levels and NF-kB expression), effect on angiogenesis (VEGF blood levels and Caspase 3 expression) and skin graft vascularization (Micro Vessels Density /MVD).
Methods: The study used 24 white male rats (Rattus norvegicus) aged 3-4 months, body weight 150-300 grams, and divided into a control group with skin graft without treatment and three groups of skin graft treated with a dosage, 200 mg/kg-BW EEP, a dosage, 400 mg/kg-BW EEP, dan a dosage, 800 mg/kgBW EEP) for 7 days. Wound tissue granulation is made according to previous research. Skin graft procedure uses allograft donors for 7 days. Blood draw from retro orbita veins and skin graft with a small amount of recipient tissue done after the seventh day. The propolis ethanol extract of mount Lawu (EEP) is made by maceration technique 3,75 liters of ethanol 70% as a solvent. Using the SPSS 22 for windows program, blood MDA levels, blood IL-6 levels, and blood VEGF levels were analyzed by One Way ANOVA and post hoc. Examination of Nuclear Factor-Kappa Beta (NF-kB) expression, Caspase-3 expression, and MVD was carried out by using Immunohistochemistry (IHC) tested with non-parametric Mann-Whitney test.
Results: There was a significant decrease in MDA blood levels (p<0.05), with an effective dosage, 400 mg/kgBW EEP and a dosage, 800 mg/kg-BW EEP. There was a significant decrease of IL-6 blood levels (p<0.05) at administration dosage, 200 mg/kg-BW EEP and a dosage, 400 mg/kg-BW EEP, and statistically significant (p<0.05). Vascular Endothelial Growth Factor (VEGF) blood levels increased with a dosage from 200 mg/kg-BW EEP until a dosage, 800 mg/kgBW EEP and significant (p<0.05), the post hoc test showed significant results in all group comparisons. There was no significant increase of skin graft vascularization (MVD) (p=0.254). Caspase-3 and NF-kB expressions decreased at certain dosages, although the decrease of Caspase-3 (p=0.557) and NF-kB (p=0.916) were not significant in statistic analysis test.
Conclusion: Propolis ethanol extract of mount Lawu (EEP) has anti-ROS, anti-inflammatory, and angiogenic effect through decreased MDA blood levels, decreased IL-6 blood levels and increased VEGF blood levels, no significant reduction in NF-kB and caspase-3 expression and there was no increase in skin graft vascularization (MVD).