探討金雀異黃酮透過雄性激素受體訊號途徑抑制蟹足腫纖維母細胞遷移的機制

中文摘要蟹足腫（keloid）是一種由傷口癒合過程中細胞外基質（extracellular matrix, ECM）過度堆積纖維母細胞大量增生並遷移。目前治療仍有限，因此需發展新的治療策略。Genistein（金雀異黃酮），具有抗發炎及抑制細胞增生的活性。本研究旨在探討Genistein對蟹足腫成纖維細胞（keloid fibroblasts, KFs）之影響。透過MTT assay分析200μM的Genistein並不會造成細胞的毒性，但Wound Healing的分析得知Genistein可顯著抑制細胞的遷移能力。本研究利用次世代定序法進行RNA sequencing分析基因變化有上升492個，下降的527個。以q-PCR偵測Genistein處理後AR (Androgen receptor)、ASPN (Asporin)的RNA表達上升，而COL1A1 (Collagen type I alpha 1 chain)、HAS2 (Hyaluronan synthase 2)、DCN (Decorin)、ANKRD1 (Ankyrin repeat domain 1)表達下降，Western Blot分析得到Genistein會促進AR的表現量且抑制COL1A1的表現。然而使用了Bicalutamide (AR抑制劑)可以明顯的逆轉 Genistein所抑制Keloid細胞的遷移能力。手術取得的keloid檢體進行體外 Genistein 注射試驗，H&E stain、免疫組織化學染色及Masson's Trichrome staining進行病理切片分析， Genistein處理後可提高AR蛋白的表現量，則可降低COL1A1蛋白表現，並減少膠原沉積。綜合上述結果，顯示Genistein具有調控蟹足腫纖維化相關分子及組織特徵之潛力，可作為蟹足腫抗纖維化治療策略之候選藥物。

Keloids are characterized by excessive extracellular matrix (ECM) deposition, fibroblast proliferation, and cell migration during the wound healing process. Current treatment options remain limited; therefore, the development of new therapeutic strategies is needed. Genistein, a naturally occurring isoflavone, possesses anti-inflammatory and anti-proliferative activities. This study aimed to investigate the effects of Genistein on keloid fibroblasts (KFs).The MTT assay demonstrated that 200 μM Genistein did not induce cytotoxicity, whereas the wound healing assay showed that Genistein significantly inhibited cell migration. RNA sequencing analysis identified 492 upregulated genes and 527 downregulated genes following Genistein treatment. Quantitative polymerase chain reaction (qPCR) analysis showed that Genistein increased the RNA expression of AR and ASPN, while decreasing the expression of COL1A1, HAS2, DCN, and ANKRD1. Western blot analysis further demonstrated that Genistein increased AR protein expression and suppressed COL1A1 protein expression. However, treatment with Bicalutamide (an AR antagonist) markedly reversed the inhibitory effect of Genistein on the migration of keloid cells. Surgically obtained keloid tissues were subjected to an ex vivo Genistein injection experiment. Histological analyses, including hematoxylin and eosin (H&E) staining, immunohistochemistry (IHC), and Masson's trichrome staining, demonstrated that Genistein increased AR protein expression, decreased COL1A1 protein expression, and reduced collagen deposition. Taken together, these results suggest that Genistein has the potential to regulate fibrosis-related molecules and histological characteristics of keloids and may serve as a potential candidate for anti-fibrotic therapy for keloids.