Arthritis and Physical Treatment

关节炎和物理治疗

• 批准号: 6467643
• 负责人: HUI Bin SUN
• 金额: $ 7.32万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-15 至 2005-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6467643
• 关键词: SDS polyacrylamide gel electrophoresis; arthritis therapy; autoimmune disorder; clinical research; collagenase; complementary DNA; fos protein; gene expression; mechanical stress; messenger RNA; polymerase chain reaction; rheumatoid arthritis; stromelysin; synovial fluid; tissue /cell culture; tissue inhibitor of metalloproteinases; transcription factor; transfection; western blottings

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.

描述（由申请人提供）：该提案的长期目标 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物理疗法来缓解疼痛和 关节炎关节僵硬。使用两种分离的人类滑膜细胞培养物 我们最近从类风湿性关节炎患者中发现机械 几dyn/cm 2 的剪切会暂时降低转录水平 基质金属蛋白酶 (MMP)-1、MMP-13 基因以及 ets-1 转录 因子，而相同的剪切会增加组织抑制剂的 mRNA 水平 金属蛋白酶 (TIMP)-1、TIMP-2 和 c-fos。这些初步基因 表达结果表明机械剪切应力的潜在用途 治疗工具并允许我们检验以下假设： 轻微机械剪切应力的非平稳时间分布 dyn/cm2 可以同时维持降低的 MMP-1、3 和 -13 mRNA 水平 以及通过下调 TIMP-1 和 2 的 mRNA 水平增加 ets-1 转录因子。 该项目的两个具体目标是 (i) 评估拟议的五个 减少 MMP rRNA 和增加的非平稳剪切应力分布 TIMP mRNA，以及 (ii) 鉴定 ets-1 对机械性能的功能 MMPs 和 TIMPs 同时调节中的应激诱导反应。我们 将从经应激处理的滑膜细胞培养物中分离 RNA 并确定 特定 MMP 和 TIMP 以及 AP-1 和 ets 基因的 cDNA 水平 使用逆转录聚合酶链式反应的家庭成员 程序。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法确定 MMP 活性。经过 转染ets-1，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存在非侵入性 可以开发物理治疗来防止组织退化 关节炎关节。