Project Details
Description
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.
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.
| Status | Finished |
|---|---|
| Effective start/end date | 8/15/02 → 7/31/07 |
ASJC
- Medicine(all)