Wang Zhen, Sun Yang, Chen Jia-Lu, Gong Yi-Ming
Department of Stomatology, Zhongshan Hospital, Fudan University. Shanghai 200032, China. E-mail:
Shanghai Kou Qiang Yi Xue. 2020 Aug;29(4):359-364.
The aim of the present study was to investigate the effect of curcumin (Cur) on TGF-β1/Smad3 pathway of rat gingival fibroblast treated with cyclosporine A (CsA) in vitro, and to provide theoretical basis for the mechanism of curcumin inhibiting drug-induced gingival hyperplasia induced by CsA.
Healthy Sprague-Dawley rat gingival fibroblasts were cultured with different concentrations of Cur (0, 5, 10, 20, 30 μmol/L) and Cur (20 μmol/L)+CsA(200 ng/mL), cell proliferation was assessed with CCK-8 assay. The mRNA levels of TGF-β1, Smad3, α-SMA and collagen type Ⅰ in gingival fibroblasts were detected by real-time PCR under Cur(20 μmol/L)+CsA(200 ng/mL); the protein level of TGF-β1, Smad3, p-Smad3, α-SMA and collagen type Ⅰ were determined through Western blot. The effect of Cur(20 μmol/L)+CsA(200 ng/mL) on migration ability of gingival fibroblasts was observed through Scratch wound-healing assay. The data were analyzed with SPSS 23.0 software package.
Cell proliferation and migration ability of rats gingival fibroblasts were significantly reduced under Cur(20 μmol/L)+CsA(200 ng/mL). 20 μmol/L Cur significantly decreased mRNA expression of TGF-β1, α-SMA and collagen type Ⅰ in gingival fibroblasts, and Western blot suggested significantly down-regulated expression of TGF-β1, p-Smad3, α-SMA, and collagen typeⅠ.
Cur may inhibit TGF-β1/Smad3 signaling pathway of gingival fibroblasts activated by CsA, thereby weakening proliferation and migration, reducing secretion of smooth muscle actin and collagen of gingival fibroblasts, and ameliorating gingival hyperplasia.
本研究旨在探讨姜黄素（Cur）对体外环孢素A（CsA）处理的大鼠牙龈成纤维细胞TGF-β1/Smad3信号通路的影响，为姜黄素抑制CsA所致药物性牙龈增生的作用机制提供理论依据。
用不同浓度的Cur（0、5、10、20、30 μmol/L）及Cur（20 μmol/L）+CsA（200 ng/mL）培养健康的Sprague-Dawley大鼠牙龈成纤维细胞，采用CCK-8法检测细胞增殖情况。在Cur（20 μmol/L）+CsA（200 ng/mL）条件下，通过实时PCR检测牙龈成纤维细胞中TGF-β1、Smad3、α-SMA和Ⅰ型胶原的mRNA水平；通过蛋白质印迹法测定TGF-β1、Smad3、p-Smad3、α-SMA和Ⅰ型胶原的蛋白水平。通过划痕伤口愈合试验观察Cur（20 μmol/L）+CsA（200 ng/mL）对牙龈成纤维细胞迁移能力的影响。数据采用SPSS 23.0软件包进行分析。
Cur（20 μmol/L）+CsA（200 ng/mL）作用下大鼠牙龈成纤维细胞的增殖和迁移能力显著降低。20 μmol/L Cur显著降低牙龈成纤维细胞中TGF-β1、α-SMA和Ⅰ型胶原的mRNA表达，蛋白质印迹法显示TGF-β1、p-Smad3、α-SMA和Ⅰ型胶原的表达明显下调。
Cur可能抑制CsA激活的牙龈成纤维细胞的TGF-β1/Smad3信号通路，从而减弱细胞增殖和迁移，减少牙龈成纤维细胞平滑肌肌动蛋白和胶原的分泌，改善牙龈增生。
