Effect of Absolute From Hibiscus syriacus L. Flower on Wound Healing in Keratinocytes

BACKGROUND: Proliferation and migration of keratinocytes are essential for the repair of cutaneous wounds. Hibiscus syriacus L. has been used in Asian medicine; however, research on keratinocytes is inadequate. OBJECTIVE: To establish the dermatological properties of absolute from Hibiscus syriacus L. flower (HSF) and to provide fundamental research for alternative medicine. MATERIALS AND METHODS: We identified the composition of HSF absolute using gas chromatography-mass spectrometry analysis. We also examined the effect of HSF absolute in HaCaT cells using the XTT assay, Boyden chamber assay, sprout-out growth assay, and western blotting. We conducted an in-vivo wound healing assay in rat tail-skin. RESULTS: Ten major active compounds were identified from HSF absolute. As determined by the XTT assay, Boyden chamber assay, and sprout-out growth assay results, HSF absolute exhibited similar effects as that of epidermal growth factor on the proliferation and migration patterns of keratinocytes (HaCaT cells), which were significantly increased after HSF absolute treatment. The expression levels of the phosphorylated signaling proteins relevant to proliferation, including extracellular signal-regulated kinase 1/2 (Erk 1/2) and Akt, were also determined by western blot analysis. CONCLUSION: These results of our in-vitro and ex-vivo studies indicate that HSF absolute induced cell growth and migration of HaCaT cells by phosphorylating both Erk 1/2 and Akt. Moreover, we confirmed the wound-healing effect of HSF on injury of the rat tail-skin. Therefore, our results suggest that HSF absolute is promising for use in cosmetics and alternative medicine. SUMMARY: Hisbiscus syriacus L. flower absolute increases HaCaT cell migration and proliferation. Hisbiscus syriacus L. flower absolute regulates phosphorylation of ERK 1/2 and Akt in HaCaT cell. Treatment with Hisbiscus syriacus L. flower induced sprout outgrowth. The wound in the tail-skin of rat was reduced by Hisbiscus syriacus L. flower absolute. Abbreviations used: HSF: Hibiscus syriacus L. flower, Erk 1/2: extracellular signal-regulated kinase 1/2, EGF: epidermal growth factor, GC/MS: gas chromatography-mass spectrometry, DMEM: dulbecco's modified eagle medium, FBS: fetal bovine serum, BSA: bovine serum albumin, p-Akt: phosphorylation of Akt, p-Erk 1/2: phosphorylation of Erk 1/2.