Simvastatin inhibits transforming growth factor-β1-induced expression of type I collagen, CTGF, and α-SMA in keloid fibroblasts
Simvastatin inhibits transforming growth factor-β1-induced expression of type I collagen, CTGF, and α-SMA in keloid fibroblasts
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DOI：
10.1111/wrr.12136
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发表时间：
2014-01-01
影响因子：
2.9
通讯作者：
Ko, Hyun-Chang
中科院分区：
文献类型：
--
作者：
Mun, Je-Ho;Kim, Young-Mi;Ko, Hyun-Chang
辛伐他汀是一种3-羟基-3-甲基戊二酰辅酶- a还原酶抑制剂，用于降低胆固醇水平。越来越多的证据表明辛伐他汀具有免疫调节和抗炎作用，可预防心血管疾病。此外，他汀类药物对各种器官纤维化的有益作用也有报道。然而，他汀类药物对瘢痕疙瘩真皮纤维化的功能作用尚未探讨。本研究的目的是确定辛伐他汀是否会影响瘢痕疙瘩相关的皮肤纤维化。我们检测了辛伐他汀对转化生长因子(TGF)- β 1诱导的I型胶原、结缔组织生长因子（CTGF或CCN2）和α -平滑肌肌动蛋白（α - sma）产生的影响。培养瘢痕疙瘩成纤维细胞，在tgf - β 1存在的情况下，将其暴露于不同浓度的辛伐他汀中，测定辛伐他汀对tgf - β 1诱导的瘢痕疙瘩成纤维细胞胶原和CTGF生成的影响。Western blotting检测I型胶原、CTGF、α - sma表达水平及Smad2、Smad3磷酸化水平。通过测定3-[4,5-二甲基噻唑-2-基]-2,5-二苯基溴化四唑的比色转化率来评估辛伐他汀对细胞活力的影响。辛伐他汀以浓度依赖的方式抑制tgf - β 1诱导的I型胶原、CTGF和α - sma的产生。辛伐他汀预处理可消除tgf - β 1诱导的Smad2和Smad3磷酸化水平。辛伐他汀对I型胶原、CTGF和α - sma表达的抑制作用被焦磷酸香叶基香叶基逆转，表明辛伐他汀诱导的细胞反应是由于抑制小GTPase Rho的参与。RhoA活化实验显示，辛伐他汀预孵育可显著阻断tgf - β 1诱导的RhoA活化。rho相关的卷曲激酶抑制剂Y27632消除了tgf - β 1诱导的I型胶原、CTGF和α - sma的产生。然而，Y27632对tgf - β 1诱导的Smad2和Smad3磷酸化无显著影响。总之，本研究提示辛伐他汀可有效抑制瘢痕疙瘩成纤维细胞中tgf - β 1诱导的I型胶原、CTGF和α - sma的产生。
Simvastatin, a 3-hydroxy-3-methylglutaryl coenzyme-A reductase inhibitor, is used to reduce cholesterol levels. Accumulating evidence has revealed the immunomodulatory and anti-inflammatory effects of simvastatin that prevent cardiovascular diseases. In addition, the beneficial effects of statins on fibrosis of various organs have been reported. However, the functional effect of statins on dermal fibrosis of keloids has not yet been explored. The objective of this study was to determine whether simvastatin could affect dermal fibrosis associated with keloids. We examined the effect of simvastatin on transforming growth factor (TGF)-beta 1-induced production of type I collagen, connective tissue growth factor (CTGF or CCN2), and alpha-smooth muscle actin (alpha-SMA). Keloid fibroblasts were cultured and exposed to different concentrations of simvastatin in the presence of TGF-beta 1, and the effects of simvastatin on TGF-beta 1-induced collagen and CTGF production in keloid fibroblasts were determined. The type I collagen, CTGF, and alpha-SMA expression levels and the Smad2 and Smad3 phosphorylation levels were assessed by Western blotting. The effect of simvastatin on cell viability was evaluated by assessing the colorimetric conversion of 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide. Simvastatin suppressed TGF-beta 1-induced type I collagen, CTGF, and alpha-SMA production in a concentration-dependent manner. The TGF-beta 1-induced Smad2 and Smad3 phosphorylation levels were abrogated by simvastatin pretreatment. The inhibition of type I collagen, CTGF, and alpha-SMA expression by simvastatin was reversed by geranylgeranyl pyrophosphate, suggesting that the simvastatin-induced cellular responses were due to inhibition of small GTPase Rho involvement. A RhoA activation assay showed that preincubation with simvastatin significantly blocked TGF-beta 1-induced RhoA activation. The Rho-associated coiled kinase inhibitor Y27632 abrogated TGF-beta 1-induced production of type I collagen, CTGF, and alpha-SMA. However, Y27632 had no significant effect on TGF-beta 1-induced phosphorylation of Smad2 and Smad3. In conclusion, the present study suggests that simvastatin is an effective inhibitor of TGF-beta 1-induced type I collagen, CTGF, and alpha-SMA production in keloid fibroblasts.