姜黄素对环孢素A诱导激活的大鼠牙龈成纤维细胞TGF-β1/Smad3通路的抑制作用

doi:10.19439/j.sjos.2020.04.005

上海口腔医学 ›› 2020, Vol. 29 ›› Issue (4): 359-364.doi: 10.19439/j.sjos.2020.04.005

姜黄素对环孢素A诱导激活的大鼠牙龈成纤维细胞TGF-β1/Smad3通路的抑制作用

王震¹, 孙扬¹, 陈佳露², 龚逸明¹

1.复旦大学附属中山医院口腔科,上海 200032;
2.普陀区眼病牙病防治所口腔科,上海 200060

收稿日期:2019-09-23 修回日期:2019-11-17 出版日期:2020-08-25 发布日期:2020-09-11
通讯作者: 龚逸明,E-mail：gongymingi@aliyun.com
作者简介:王震（1994-）,男,在读硕士研究生,E-mail：2212204835@qq.com

Effect of curcumin on TGF-β1 /Smad3 pathway in rat gingival fibroblast treated with cyclosporine A

WANG Zhen¹, SUN Yang¹, CHEN Jia-lu², GONG Yi-ming¹

1. Department of Stomatology, Zhongshan Hospital, Fudan University. Shanghai 200032;
2. Department of Stomatology, Ophthalmic & Dental Center of Putuo District. Shanghai 200060, China

Received:2019-09-23 Revised:2019-11-17 Online:2020-08-25 Published:2020-09-11

摘要/Abstract

摘要： 目的：体外实验研究姜黄素（curcumin,Cur）对环孢素A（cyclosporine A,CsA）作用大鼠牙龈成纤维细胞TGF-β1/Smad3通路的影响,为进一步探讨Cur抑制CsA所致药物性牙龈增生的发生机制提供理论依据。方法：使用CCK-8法观察0、5、10、20、30 μmol/L Cur对大鼠牙龈成纤维细胞增殖的影响, 20 μmol/L Cur+200 ng/mL CsA共同作用对大鼠牙龈成纤维细胞增殖的影响。实时荧光定量PCR检测20 μmol/L Cur+200 ng/mL CsA共同作用下,牙龈成纤维细胞中转化生长因子TGF-β1、Smad3、平滑肌肌动蛋白α-SMA和I型胶原蛋白COL-I的mRNA表达变化;蛋白免疫印迹实验检测TGF-β1、Smad3、p-Smad3、α-SMA和COL-I的蛋白表达变化。细胞划痕实验观察20 μmol/L Cur+200 ng/mL CsA 对牙龈成纤维细胞迁移能力的影响。采用SPSS 23.0 软件包对数据进行统计学分析。结果：大鼠牙龈成纤维细胞在20 μmol/L Cur+200 ng/mL CsA共同作用下,细胞增殖和迁移能力明显降低;20 μmol/L Cur显著下调了牙龈成纤维细胞中TGF-β1、α-SMA 和COL-I的mRNA 表达,蛋白免疫印迹实验提示,TGF-β1、p-Smad3、α-SMA 和COL-I的表达同样显著下调。结论：Cur可能通过抑制CsA激活的牙龈成纤维细胞TGF-β1/Smad3 信号通路,从而降低牙龈成纤维细胞增殖、迁移、平滑肌肌动蛋白和胶原分泌,改善牙龈增生。

关键词: 姜黄素, 环孢素A, 牙龈成纤维细胞, TGF-β1, Smad3, 牙龈增生

Abstract: PURPOSE: 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. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

Key words: Curcumin, Cyclosporine A, Gingival fibroblasts, TGF-β1, Smad3, Gingival hyperplasia

中图分类号:

R781.1

王震, 孙扬, 陈佳露, 龚逸明. 姜黄素对环孢素A诱导激活的大鼠牙龈成纤维细胞TGF-β1/Smad3通路的抑制作用[J]. 上海口腔医学, 2020, 29(4): 359-364.

WANG Zhen, SUN Yang, CHEN Jia-lu, GONG Yi-ming. Effect of curcumin on TGF-β1 /Smad3 pathway in rat gingival fibroblast treated with cyclosporine A[J]. Shanghai Journal of Stomatology, 2020, 29(4): 359-364.

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姜黄素对环孢素A诱导激活的大鼠牙龈成纤维细胞TGF-β1/Smad3通路的抑制作用

Effect of curcumin on TGF-β1 /Smad3 pathway in rat gingival fibroblast treated with cyclosporine A

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