Arthritis and Physical Treatment

描述（由申请人提供）：本提案的长期目标 旨在阐明机械刺激对组织降解的影响， 类风湿性关节炎并开发一种缓解疼痛和 关节炎关节僵硬。使用两种分离的人类滑膜细胞培养物 从类风湿性关节炎患者，我们最近发现， 在几个dyn/cm 2的剪切瞬时降低转录水平， 基质金属蛋白酶（MMP）-1、MMP-13基因以及ets-1转录 因子，而相同的剪切增加组织抑制因子的mRNA水平。 TIMP-1、TIMP-2和c-fos的表达。这些初步的基因 表达结果表明，机械剪切应力作为一个潜在的用途， 治疗工具，并允许我们测试以下假设：一个适当的 非平稳的时间曲线的温和的机械剪切应力在几个 dyn/cm 2可同时维持MMP-1、3和-13 mRNA水平的降低 TIMP-1和TIMP-2的mRNA水平也通过下调 ets-1转录因子的表达。 该项目的两个具体目标是：（一）评估拟议的五个 MMP rRNA减少和MMP rRNA增加的非稳定剪切应力曲线 TIMP mRNAs，以及（ii）鉴定ets-1在机械调节中的功能。 应激诱导的MMPs和TIMPs的同时调节反应。我们 将从应激处理的滑膜细胞培养物中分离RNA， 特异性MMPs和TIMPs以及AP-1和ets基因的cDNA水平 使用逆转录-聚合酶链反应 procedure.我们还将通过免疫印迹法测量MMP蛋白的水平， 通过使用酶谱法和原纤维降解测定来确定MMP活性。通过 通过对ets-1的测试，我们将在机械条件下测试ets-1的功能。 刺激。拟议的项目将有助于回答是否是一个非侵入性的 可以开发物理治疗以防止组织降解， 关节炎

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.