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. 2019 Aug 24;20(17):4134. doi: 10.3390/ijms20174134

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© 2019 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Apoptosis and autophagy assay of glycyrrhizin-treated keloids. (a,b) The annexin V-FITC assay showed that glycyrrhizin induced apoptosis in HDFs and KFs (*** p < 0.001). (c) Representative images of keloid spheroids stained by TUNEL for apoptosis assay. The results showed significantly enhanced apoptosis in glycyrrhizin-treated keloid spheroids, and TUNEL-positive cells dose-dependently increased. Data are expressed as mean ± SEM of five experiments (*** p < 0.001). (d) Western blot analysis of autophagy markers in HDFs, KFs, and KFs treated with 200 µM of glycyrrhizin. Beclin 1 and LC3 levels were significantly increased in KFs (* p < 0.05). The autophagy marker LC3 showed a decreased expression after glycyrrhizin treatment in KFs (** p < 0.01). (e) IHC for autophagy markers in keloid tissue after glycyrrhizin treatment (0, 200, 500 µM). Semi-quantitative analysis revealed that the expression levels of Beclin 1 and LC3 were dose-dependently reduced in glycyrrhizine-treated keloid tissue (*** p < 0.001).