Prevention of Simvastatin-Induced Inhibition of Tendon Cell Proliferation and Cell Cycle Progression by Geranylgeranyl Pyrophosphate
Prevention of Simvastatin-Induced Inhibition of Tendon Cell Proliferation and Cell Cycle Progression by Geranylgeranyl Pyrophosphate
复制标题
DOI：
10.1093/toxsci/kfv239
复制
发表时间：
2016-02-01
影响因子：
3.8
通讯作者：
Pang, Jong-Hwei S.
中科院分区：
文献类型：
--
作者：
Tsai, Wen-Chung;Yu, Tung-Yang;Pang, Jong-Hwei S.
据报道，他汀类药物可引起肌腱病变，甚至肌腱断裂。本研究旨在探讨辛伐他汀对肌腱细胞不良作用的分子机制。使用从大鼠跟腱分离的肌腱细胞进行体外肌腱愈合模型。MTT法检测细胞活力，流式细胞仪检测细胞周期。Ki-67免疫荧光染色用于评估肌腱细胞的增殖活性。Western blot和免疫共沉淀法检测细胞周期相关蛋白的表达。为了研究他汀类药物对肌腱细胞作用的潜在机制，将甲羟戊酸、焦磷酸法呢酯（FPP）或焦磷酸香叶基香叶基酯（GGPP）加入辛伐他汀处理的肌腱细胞中。辛伐他汀对体外肌腱愈合模型和肌腱细胞增殖有抑制作用，且呈剂量依赖性。免疫荧光染色显示辛伐他汀处理的肌腱细胞Ki-67表达减少。此外，辛伐他汀诱导细胞周期停滞在G1期。辛伐他汀以剂量依赖性方式下调cdk 1、cdk 2、细胞周期蛋白A和细胞周期蛋白E的表达水平。辛伐他汀的抑制作用被证明介导了甲羟戊酸的减少，外源性GGPP的加入完全阻止了辛伐他汀对肌腱细胞的抑制作用。本研究首次证实了辛伐他汀诱导的肌腱病变或肌腱断裂的分子机制。GGPP被证明可以防止辛伐他汀对肌腱细胞的不良影响，而不会干扰其降低胆固醇的功效。
Statins have been reported to induce tendinopathy and even tendon rupture. The present study was designed to investigate the potential molecular mechanism underlying the adverse effect of simvastatin on tendon cells. An in vitro tendon healing model was performed using tendon cells isolated from rat Achilles tendons. The viability of tendon cells and cell cycle progression were examined by the MTT assay and flow cytometric analysis, respectively. Immunofluorescent staining for Ki-67 was used to assess the proliferation activity of tendon cells. Western blot analysis and coimmunoprecipitation was used to determine the protein expression of cell cycle-related proteins. To investigate the potential mechanism underlying the effect of statins on tendon cells, mevalonate, farnesyl pyrophosphate (FPP), or geranylgeranyl pyrophosphate (GGPP) was added to simvastatin-treated tendon cells. Simvastatin inhibited the in vitro tendon healing model and tendon cell proliferation in a dose-dependent manner. Immunofluorescent staining demonstrated reduced ki-67 expression in simvastatin-treated tendon cells. Furthermore, simvastatin induced cell cycle arrest at the G1 phase. The expression levels of cdk1, cdk2, cyclin A, and cyclin E were downregulated by simvastatin in a dose-dependent manner. The inhibitory effect of simvastatin was proved to mediate the reduction of mevalonate, and the addition of exogenous GGPP completely prevented the inhibitory effect of simvastatin on tendon cells. The present study demonstrated, for the first time, the molecular mechanism underlying simvastatin-induced tendinopathy or tendon rupture. GGPP was shown to prevent the adverse effect of simvastatin in tendon cells without interfering with its cholesterol-reducing efficacy.