Arthritis and Physical Treatment

描述(由申请人提供)：本计划的长期目标 是为了阐明机械刺激对组织降解的影响 类风湿性关节炎和开发一种物理疗法来缓解疼痛和 关节炎关节僵硬。用分离的两种人滑膜细胞培养物 从类风湿性关节炎患者中，我们最近发现机械性 以几达因/厘米2的剪切率瞬时降低了转录水平 基质金属蛋白酶-1、13基因与Ets-1转录 因子，而相同的剪切力增加了组织抑制物的mRNA水平 金属蛋白酶(TIMP)-1、TIMP-2和c-fos的表达。这些初步基因 表达结果表明，机械剪应力可以作为一种潜在的 治疗工具，并允许我们检验以下假设：一个适当的 几种情况下机械剪切应力的非平稳时间分布 Dyn/cm2可同时维持降低的基质金属蛋白酶-1、3和-13的mRNA水平 以及通过下调TIMP-1和TIMP-2的mRNA水平 Ets-1转录因子的表达。 本项目的两个具体目标是：(一)评估拟议的五个 减少和增加基质金属蛋白酶rRNA的非平稳剪应力剖面 TIMP mRNAs，以及(Ii)鉴定ETS-1在机械反应中的功能 MMPs和TIMPs同时调节中的应激反应。我们 将从应激处理的滑膜细胞培养物中分离出RNA并确定 特异性MMPs和TIMPs以及AP-1和ETS基因的cDNA水平 利用逆转录-聚合酶链式反应检测家庭成员 程序。我们还将通过免疫印迹和 用酶谱和纤维降解试验测定基质金属蛋白酶活性。通过 将ETS-1基因导入Ets-1，检测Ets-1在机械状态下的功能 刺激物。建议的项目将有助于回答非侵入性 可以开发物理治疗来防止组织退化 关节炎关节。

DESCRIPTION (provided by applicant): The long-term objectives of this proposal are to elucidate the effects of mechanical stimuli to tissue degradation of rheumatoid arthritis and to develop a physical treatment for relieving pain and stiffness of arthritic joints. Using two human synovial cell cultures isolated from rheumatoid arthritis patients, we have recently found that mechanical shearing at a few dyn/cm 2 transiently decreases the transcriptional levels of matrix metalloproteinase (MMP)-1, MMP-13 genes as well as ets-1 transcription factor, while the same shearing increases the mRNA levels of tissue inhibitor of metalloproteinase (TIMP)-1, TIMP-2 and c-fos. These preliminary gene expression results suggest a potential use of mechanical shear stress as a therapeutic tool and allow us to test the following hypothesis: An appropriate non-stationary temporal profile of gentle mechanical shear stress at a few dyn/cm2 can maintain simultaneously a reduced mRNA level of MMP-1, 3, and -13 as well as an increased mRNA level of TIMP-1 and 2 through the down-regulation of ets-1 transcription factor. Two specific aims of this project are (i) to evaluate the proposed five non-stationary shear stress profiles for decreasing MMP rRNAs and increasing TIMP mRNAs, and (ii) to identify the function of ets-1 on mechanical stress-induced response in the simultaneous regulation of MMPs and TIMPs. We will isolate RNA from the stress-treated synovial cell cultures and determine the cDNAs levels of the specific MMPs and TIMPs as well as AP-1 and ets gene family members using a reverse transcription-polymerase chain reaction procedure. We will also measure the level of MMP proteins by immunoblotting and determine MMP activities by using zymography and a fibril degradation assay. By transfecting ets-1, we will test the function of ets-1 under mechanical stimuli. The proposed project will contribute to answer whether a non-invasive physical treatment can be developed for preventing from tissue degradation in arthritis joints.