Anti-arthritic effects of combined treatment with histone deacetylase inhibitor and low-intensity ultrasound in the presence of microbubbles in human rheumatoid synovial cells
Anti-arthritic effects of combined treatment with histone deacetylase inhibitor and low-intensity ultrasound in the presence of microbubbles in human rheumatoid synovial cells
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DOI：
10.1093/rheumatology/ken003
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发表时间：
2008-04-01
期刊：
影响因子：
5.5
通讯作者：
Kimura, T.
中科院分区：
文献类型：
--
作者：
Nakamura, C.;Matsushita, I.;Kimura, T.
目标。观察组蛋白脱乙酰酶(HDAC)抑制剂联合超声(1 MHz，10%占空比，0.1或0.2W/cm(2))对RA滑膜成纤维细胞(RASF)的作用。RASF是从全膝关节置换术中RA患者的类风湿滑膜组织中分离出来的。RASF用HDAC抑制剂曲古抑素A(TSA)处理，加或不加US。台盼蓝拒染法检测细胞活力，碘化丙啶(PI)染色流式细胞仪检测细胞周期。半定量RT-PCR检测细胞周期相关基因Cyclin D、Cyclin A、Cyclin B和p21(WAF1/Cip1)的基因表达。用Annexin V、FITC和PI染色的流式细胞仪检测细胞的凋亡。应用基因芯片技术分析炎症相关基因的表达。观察到TSA对RASF细胞活力、细胞周期停滞和细胞凋亡的影响呈剂量依赖性下降。在存在微泡的情况下，US治疗增加了细胞的摄取，但不会导致细胞周期停滞或细胞凋亡。TSA和US联合应用可调控细胞周期相关基因的表达，显著减少S期细胞数，增加G(2)-M期细胞数。US还进一步增强了TSA诱导的RASF细胞凋亡和炎症相关基因的调控表达。HDAC抑制剂与US联合应用可有效降低RASF的细胞存活率并诱导其凋亡。综合疗法对控制滑膜增殖和炎症是有用的，因为US可以很容易地作为局部理疗应用于靶点关节。
Objective. The therapeutic effects of histone deacetylase (HDAC) inhibitor combined with ultrasound (US) (1 MHz, 10% duty factor, 0.1 or 0.2 W/cm(2)) in RA synovial fibroblasts (RASFs) were examined.Methods. RASFs were isolated from rheumatoid synovial tissues obtained from patients with RA during total knee arthroplasty. RASFs were treated with an HDAC inhibitor, trichostatin A (TSA), with or without US. Cell viability was estimated using the Trypan blue dye exclusion test and cell cycle was examined by flow cytometry using propidium iodide (PI) staining. Gene expression of cell cycle-related genes cyclin D, cyclin A, cyclin B and p21(WAF1/Cip1) was analysed by semi-quantitative RT-PCR. Detection of apoptosis was examined by flow cytometry using annexin V FITC and PI staining. Microarray analysis was carried out to profile gene expression of inflammation-related genes.Results. Dose-dependent decreases in cell viability, cell cycle arrest and apoptosis in RASFs due to TSA were observed. US treatment in the presence of microbubbles increased cellular uptake, but did not induce cell cycle arrest or apoptosis. The combination of TSA and US modulated cell cycle-related gene expression and significantly decreased S phase cells and increased G(2)-M phase cells. US also further enhanced TSA-induced RASF apoptosis and regulated expression of inflammation-related genes.Conclusions. HDAC inhibitor in combination with US effectively reduces cell viability and induces apoptosis in RASFs. I he combination therapy could be useful to control synovial proliferation and inflammation, since US can be easily applied to targeted joints as local physiotherapy.