Louneva Natalia, Huaman Gonzalo, Fertala Joanna, Jiménez Sergio A
Thomas Jefferson University, Division of Rheumatology, Philadelphia, Pennsylvania 19107-5541, USA.
Arthritis Rheum. 2006 Apr;54(4):1298-308. doi: 10.1002/art.21723.
To examine whether statins are capable of modulating collagen gene expression in cultured systemic sclerosis dermal fibroblasts.
Cultured dermal fibroblasts from 3 patients with diffuse systemic sclerosis of recent onset were treated with 5 microM and 10 microM of simvastatin for 3 or 4 days. Morphologic features, cytotoxicity, and type I collagen production and messenger RNA (mRNA) levels in the fibroblasts were examined. The effects of mevalonate, geranylgeranyl pyrophosphate (GGPP), and farnesyl pyrophosphate (FPP), which are lipids downstream from the hydroxymethylglutaryl-coenzyme A block, were also examined. Transient transfections with COL1A1 promoter-reporter constructs and electrophoretic gel mobility shift assays were utilized to examine COL1A1 transcription and Sp1 and CCAAT-box binding factor (CBF) binding.
Simvastatin did not cause morphologic changes or cytotoxicity in the fibroblasts, even after 4 days of treatment. Type I collagen production and mRNA levels showed a potent and dose-related inhibition following 3 and 4 days of treatment. The inhibition of collagen gene expression by simvastatin was completely reversed by mevalonate and GGPP, but not by FPP. The statin effects occurred at the transcriptional level and involved the proximal COL1A1 promoter region encompassing -174 bp. A significant reduction in Sp1 and CBF binding activity was also found in simvastatin-treated cells.
Simvastatin is a powerful inhibitor of type I collagen gene expression in normal and systemic sclerosis fibroblasts. The pleiotropic protective effects of statins on various endothelial and immune cell functions in conjunction with their potent inhibitory effects on type I collagen gene expression suggest that statins may be effective therapeutic agents in systemic sclerosis.
研究他汀类药物是否能够调节培养的系统性硬化症皮肤成纤维细胞中的胶原蛋白基因表达。
用5微摩尔/升和10微摩尔/升的辛伐他汀处理3例近期发病的弥漫性系统性硬化症患者的培养皮肤成纤维细胞3或4天。检测成纤维细胞的形态学特征、细胞毒性、I型胶原蛋白产生及信使核糖核酸（mRNA）水平。还检测了3-羟基-3-甲基戊二酰辅酶A阻断下游的脂质甲羟戊酸、香叶基香叶基焦磷酸（GGPP）和法尼基焦磷酸（FPP）的作用。利用COL1A1启动子-报告基因构建体进行瞬时转染和电泳凝胶迁移率变动分析，以检测COL1A1转录以及Sp1和CCAAT盒结合因子（CBF）结合情况。
即使在处理4天后，辛伐他汀也未引起成纤维细胞的形态学改变或细胞毒性。处理3天和4天后，I型胶原蛋白产生及mRNA水平呈现出强效且与剂量相关的抑制作用。辛伐他汀对胶原蛋白基因表达的抑制作用被甲羟戊酸和GGPP完全逆转，但未被FPP逆转。他汀类药物的作用发生在转录水平，且涉及包含-174碱基对的COL1A1启动子近端区域。在经辛伐他汀处理的细胞中还发现Sp1和CBF结合活性显著降低。
辛伐他汀是正常和系统性硬化症成纤维细胞中I型胶原蛋白基因表达的强效抑制剂。他汀类药物对各种内皮细胞和免疫细胞功能具有多效性保护作用，同时对I型胶原蛋白基因表达具有强效抑制作用，这表明他汀类药物可能是系统性硬化症的有效治疗药物。
