Induction of keratinocyte migration by ECa 233 is mediated through FAK/Akt, ERK, and p38 MAPK signaling

Abstract

Centella asiatica is widely considered the most important medicinal plant for treating and relieving skin diseases. Recently developed standardized extract of Centella asiatica ECa 233 has demonstrated positive effects on wound healing of incision and burn wound in rats. However, knowledge associated with wound healing mechanism of ECa 233 was scare. Therefore, this study aimed to investigate the effect and underlying molecular mechanisms of ECa 233 on the migration of a human keratinocyte cell line (HaCaT) using scratch wound healing assay. Formation of filopodia, a key protein in cell migration as well as signaling pathways possibly involved were subsequently assessed. It was found that HaCaT cell migration was significantly enhanced by ECa 233 in a concentration- and time-dependent manner. The filopodia formations were accordingly increased in exposure to ECa 233 at concentrations of 0.1-100 μg/ml. Furthermore, ECa 233 was found to significantly upregulate the expression of Rac1 and RhoA and to induce phosphorylation of FAK and Akt as well as ERK and p38 MAPK. Taken all together, it is suggestive that ECa 233 induces cell migration and subsequently promotes wound healing activity, through the activation of FAK, Akt, and MAPK signaling pathways thereby supporting the role of ECa 233 to be further developed for the clinical treatment of wound.

Citrus sudachi Peel Extract Suppresses Cell Proliferation and Promotes the Differentiation of Keratinocytes through Inhibition of the EGFR–ERK Signaling Pathway

“…The ERK pathway (Ras–Raf–MEK–ERK) is involved in the regulation of various cellular processes, such as adhesion, proliferation, cell cycle progression, migration, survival, differentiation, metabolism, and transcription. Previous studies showed that many extracts exert various biological functions by modulating the ERK pathway [ 25 , 26 ]. For example, carrot pentane-based fractions inhibited the proliferation of tumorigenic HaCaT cells through inhibition of the ERK and phosphatidylinositol 3-kinase–Akt pathways, and the extract of Centella asiatica induced keratinocyte migration through focal adhesion kinase–Akt, ERK, and p38 MAPK signaling.…”

ECa 233 Suppresses LPS-Induced Proinflammatory Responses in Macrophages <i>via</i> Suppressing ERK1/2, p38 MAPK and Akt Pathways

“…The contents of ECa 233 contained madecassoside (52%) and asiaticoside (41%) which was quantified by HPLC and liquid chromatography-tandem mass spectrometry (LC-MS/MS) as previous reported. 13,14) 2′,7′-Dichlorofluorescein diacetate (DCFH-DA), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), bovine serum albumin (BSA), dexamethasone and LPS from Escherichia coli 055:B5 were purchased from Sigma (St. Louis, MO, U.S.A.). The primary antibodies specific to cyclooxygenase-2 (COX-2), β-actin and the secondary antibody goat anti-mouse immunoglobulin G/horseradish peroxidase (IgG/HRP) were obtained from Thermo Scientific (Waltham, MA, U.S.A.).…”

“…Its major bioactive composition is madecassoside and asiaticoside. 11) The pharmacological activity of C. asiatica has been shown to exhibit antioxidant, anti-inflammatory, 12) neuroprotective 13,14) and wound healing effects. 15) A difficulty of the crude extracts of C. asiatica is its compound variation, leading to inconsistency of pharmacological responses.…”

“…11) The pharmacological activity of C. asiatica has been shown to exhibit antioxidant, anti-inflammatory, 12) neuroprotective 13,14) and wound healing effects. 15) A difficulty of the crude extracts of C. asiatica is its compound variation, leading to inconsistency of pharmacological responses. 16) An attempt to establish a pharmacological standard of the crude extract as a standardized extract of C. asiatica ECa 233 has been made to circumvent this problem.…”

“…The proportion of madecassoside and asiaticoside were kept in 1.5 ± 0.5. 14,16,17) Previous work has addressed the beneficial effect of ECa 233 on wound healing 15) and skin inflammation (unpublished data); however, the effect on inflammatory responses remains an issue of ambiguity. Therefore, the present study aims to investigate the anti-inflammatory effect of ECa 233 and the mechanism of action against LPS-induced macrophages.…”