Apoptosis of rheumatoid synovial cells by statins through the blocking of protein geranylgeranylation - A potential therapeutic approach to rheumatoid arthritis
Apoptosis of rheumatoid synovial cells by statins through the blocking of protein geranylgeranylation - A potential therapeutic approach to rheumatoid arthritis
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DOI：
10.1002/art.21564
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发表时间：
2006-02-01
影响因子：
--
通讯作者：
Minota, S
中科院分区：
文献类型：
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作者：
Nagashima, T;Okazaki, H;Minota, S
目标。目的观察他汀类药物是否能诱导类风湿关节炎（RA）滑膜细胞凋亡。研究了亲脂性和亲水性他汀类药物氟伐他汀和普伐他汀对体外培养RA滑膜细胞凋亡的影响。JC-1染色后流式细胞术分析细胞凋亡。（测量线粒体跨膜电位），活性半胱天冬酶3，膜联蛋白V和碘化丙啶。通过加回实验来确定甲羟戊酸途径的哪些下游产物可以抑制细胞凋亡。他汀类药物诱导的多种信号通路的调节，包括蛋白质戊烯酰化，也被研究。氟伐他汀而非普伐他汀诱导RA滑膜细胞凋亡呈浓度依赖性（1-1.0 μ M）和时间依赖性（48-96小时）。另一种亲脂性他汀，匹伐他汀，表现出与氟伐他汀几乎相同的效果。与此形成鲜明对比的是，亲脂性他汀类药物并没有显著增加骨关节病变患者滑膜细胞的凋亡。氟伐他汀诱导的细胞凋亡是线粒体和半胱天冬酶3依赖性的，甲羟戊酸钠和焦磷酸香叶基可消除细胞凋亡，焦磷酸法尼基不能消除细胞凋亡。香叶基香叶转移酶抑制剂GGTI-298模拟氟伐他汀对RA滑膜细胞的影响。RhoA激酶抑制剂Y-27632治疗RA滑膜细胞可引起细胞凋亡。氟伐他汀降低了RhoA蛋白在膜部分的含量，而增加了细胞质部分的含量。氟伐他汀通过线粒体和caspase 3依赖性途径以及阻断甲羟戊酸途径，特别是通过抑制蛋白香叶酰化和RhoA/RhoA激酶途径，诱导RA滑膜细胞凋亡。这些发现表明，亲脂性他汀类药物有潜力成为治疗类风湿性关节炎的新型药物。
Objective. To determine whether statins induce apoptosis in rheumatoid arthritis (RA) synoviocytes.Methods. The effects of lipophilic and hydrophilic statins (fluvastatin and pravastatin, respectively) on the apoptosis of cultured RA synoviocytes were examined in vitro. Apoptosis was analyzed by flow cytometry after staining with JC-1. (to measure the mitochondrial transmembrane potential), active caspase 3, annexin V, and propidium iodide. Add-back experiments were conducted to determine which downstream products of the mevalonate pathway could suppress apoptosis. Modulation of various signaling pathways induced by statins, including protein prenylation, was also investigated.Results. Fluvastatin, but not pravastatin, induced apoptosis in RA synoviocytes in a concentration-dependent (1-1.0 mu M) and time-dependent (48-96 hours) manner. Another lipophilic statin, pitavastatin, displayed almost the same effects as fluvastatin. In sharp contrast lipophilic statins did not significantly increase apoptosis in synoviocytes from patients with ostcoarthropathy. Apoptosis induced by fluvastatin was mitochondrial- and caspase 3-dependent and was abrogated by mevalonate and geranylgeranyl pyrophosphate, but not by farnesyl pyrophosphate. In addition, the geranylgeranyl transferase inhibitor GGTI-298 mim-icked the effect of fluvastatin on RA synoviocytes. Treatment of RA synoviocytes with the RhoA kinase inhibitor Y-27632 caused apoptosis. Fluvastatin decreased the amount of RhoA protein in the membrane fraction, but increased the amount in the cytosolic fraction.Conclusion. Fluvastatin induced apoptosis in RA synoviocytes through a mitochondrial- and caspase 3-dependent pathway and by the blockage of mevalonate pathways, particularly through the inhibition of protein geranylgeranylation and RhoA/RhoA kinase pathways. These findings suggest that lipophilic statins have potential as novel therapeutic agents for RA.