Abstract

Background Wubeizi (Rhus chinensis Mill.) ointment has been shown as an effective treatment for keloids. However, the protective mechanisms of Wubeizi ointment are not fully understood. The mammalian target of rapamycin (mTOR) has been demonstrated to be associated with keloid pathogenesis. In the present study, we investigated if Wubeizi ointment suppressed keloid formation through the modulation of key molecules of the rapamycin (mTOR) pathway including phosphatase and tensin homolog (PTEN), phosphatidylinositol 3-kinase (PI3K), and protein kinase B (Akt). Methods A keloid mouse model and human keloid-derived fibroblasts were developed and treated with Galla chinensis. Immunohistochemistry, western blot, and reverse transcription-PCR were used to detect PI3K, PTEN, Akt, and mTOR in keloid tissues and keloid fibroblasts. The apoptosis and proliferation rate of keloid fibroblasts was, respectively, analyzed by flow cytometry according to the MTT assay. Statistical analysis was done using SPSS version 20.0. For two variable comparisons, a two independent samples t-test was used. For multiple variable comparisons, data were analyzed by one-way analysis of variance (ANOVA) followed by pairwise q-tests. Results Our in vivo and in vitro studies showed that Wubeizi ointment suppressed keloid formation through inhibition of fibroblast proliferation and promotion of fibroblast apoptosis. The underlying basis involves downregulation of p-Akt and p-mTOR as well as upregulation of PTEN. Conclusion These findings may contribute to a better understanding of the mechanisms of Wubeizi ointment for treating keloids.

Highlights

  • Keloids are formed because of abnormal overgrowth of scar tissue during the healing of skin wounds and are characterized by the proliferation of dermal fibroblast and the aberrant accumulation of extracellular matrix collagens, mucins, and glycosaminoglycans [1]

  • The results of immunohistochemistry showed that most of the p-phosphatidylinositol 3-kinase (PI3K), p-Akt, and p-mammalian target of rapamycin (mTOR) were expressed in cytoplasm and a small amount of p-PI3K, p-Akt, and p-mTOR was observed in nucleus (Figure 2)

  • There was no significant difference between the Wubeizi ointment-treated and control group in PI3K mRNA levels in keloid fibroblasts (Figure 5(a), P = 0:418)

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Summary

Introduction

Keloids are formed because of abnormal overgrowth of scar tissue during the healing of skin wounds and are characterized by the proliferation of dermal fibroblast and the aberrant accumulation of extracellular matrix (type I and III) collagens, mucins, and glycosaminoglycans [1]. The mechanisms underlying the inhibitory effect of Wubeizi ointment on keloid formation are not fully understood. The expression of transforming growth factor-β (TGF-β1), PI3K, Akt, and mTOR was significantly increased in pathological scar fibroblasts than in normal skin. We investigated if Wubeizi ointment suppressed keloid formation through the modulation of key molecules of the rapamycin (mTOR) pathway including phosphatase and tensin homolog (PTEN), phosphatidylinositol 3-kinase (PI3K), and protein kinase B (Akt). Immunohistochemistry, western blot, and reverse transcription-PCR were used to detect PI3K, PTEN, Akt, and mTOR in keloid tissues and keloid fibroblasts. Data were analyzed by one-way analysis of variance (ANOVA) followed by pairwise q-tests. Our in vivo and in vitro studies showed that Wubeizi ointment suppressed keloid formation through inhibition of fibroblast proliferation and promotion of fibroblast apoptosis. These findings may contribute to a better understanding of the mechanisms of Wubeizi ointment for treating keloids

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