Naringenin potentiated β-sitosterol healing effect on the scratch wound assay

Abstract

In the present investigation scratch wound assay was used to study the ability of several combinations of each flavonoid (chrysin, naringenin or resveratrol) with β-sitosterol to heal wounds in vitro. MTT test was performed to determine if the combination of flavonoid with β-sitosterol was toxic to fibroblasts or not. Also, superoxide dismutase (SOD) activity and interleukin-1β (IL-1β) concentrations were measured. The best closure rates were obtained with β-sitosterol combined with naringenin and β-sitosterol combined with resveratrol. The combination that produced the best closure rate namely β-sitosterol with naringenin increased SOD activity significantly. However, this combination was not better than naringenin or β-sitosterol alone in reducing IL-β concentration. The results of MTT test indicated that the combination as well as β-sitosterol alone or naringenin alone has no toxic effect on fibroblasts. In conclusion, the combination of β-sitosterol and naringenin exerted a synergistic effect on wound closure without decreasing the viability of fibroblasts, increased antioxidant defense mechanism and decreased IL-β.

Keywords: Beta-sitosterol; Interleukin-1β; Naringenin; SOD; Scratch wound assay; Synergism.

Fig. 1

Scratch wound assay. (A) Negative control at zero time, (B) negative control after 24 h, (C) β-sitosterol at 50 μM after 24 h, (D) β-sitosterol at 100 μM after 24 h, (E) naringenin at 50 μM after 24 h, (F) naringenin at 100 μM after 24 h, (G) naringenin and β-sitosterol at 50 μM after 24 h, (H) naringenin and β-sitosterol at 100 μM after 24 h and (I) platelet-derived growth factor at 10 nM after 24 h.

Fig. 2

Evaluation of antoproliferative effects of β-sitosterol, naringenin or their combination on fibroblasts using MTT assay.

Fig. 3

Effect of different treatments on levels of SOD. In all groups except the first one (no wound), a linear wound on monolayer cells was created using a sterile 100 μL plastic pipette tip. *** Indicates significant differences in comparison with no treatment group (P < 0.001). ##P < 0.01 and ###P < 0.001 show significant differences between indicated groups. SOD, superoxidae dismutase

Fig. 4

Effect of different treatments on level of IL-1β. In all groups except the first one (no wound), a linear wound on monolayer cells was created using a sterile 100 μL plastic pipette tip. *** Indicates significant differences in comparison with no treatment group (P < 0.001). Interleukin-1β, IL-1β.