ABSTRACT
<p><b>OBJECTIVE</b>To investigate the role of natural killer-22 (NK-22) cells in the synovial fluid in the proliferation of fibroblast-like synoviocytes (FLS) in patients with rheumatoid arthritis (RA) and explore the possible signal pathway involved.</p><p><b>METHODS</b>NK-22 cells in the SF of RA patients were sorted by flow cytometry. NK-22 cells were cultured for two weeks and the purity was detected by flow cytometry before stimulation with 20 ng/ml phorbol 12-myristate 13-acetate and 0.5 µmol/L ionomycin for 4 h. The level of interleukin-22 (IL-22) in the culture medium supernatant was then measured with ELISA. The proliferation of FLS in the presence of the culture supernatant of NK-22 cells was assessed with MTT assay at 24, 48 and 72 h, and the effect of IL-22 antibody on FLS proliferation was also observed. Real-time PCR and Western blotting were used to detect Stat3 mRNA and p-Stat3 protein levels, respectively, in the FLS exposed to rhIL-22 and AG490.</p><p><b>RESULTS</b>NK-22 cells were successfully sorted by flow cytometry with a purity exceeding 90%. The levels of IL-22 in the supernatant of NKp44(+)NK cell culture averaged 1273.42∓254.48 pg/ml. The FLS proliferated rapidly 24, 48, and 72 h after the addition of culture supernatant of NK-22 cells (P<0.05). IL-22 antibody obviously inhibited the proliferation of FLS induced by NK-22 cell culture supernatant (P<0.05). Exposure of the FLS to rhIL-22 obviously increased cellular Stat3 expression levels, which were significantly lowered by the addition of AG490 (P<0.05).</p><p><b>CONCLUSION</b>NK-22 cells in the SF of RA patients can produce high concentrations of IL-22 to promote the proliferation of FLS through the STAT3 signal pathway.</p>
Subject(s)
Humans , Arthritis, Rheumatoid , Metabolism , Cell Proliferation , Cells, Cultured , Fibroblasts , Cell Biology , Interleukins , Metabolism , Killer Cells, Natural , Cell Biology , Metabolism , STAT3 Transcription Factor , Metabolism , Signal Transduction , Synovial Fluid , Cell Biology , Synovial Membrane , Cell BiologyABSTRACT
Objective: To investigate the effects of point thread embedding on the learning and memory of the vascular dementia(VD) model rats and the possible mechanisms. Methods: VD rat models were established by employing improved method of Pulsinelli's four-vessel occlusion,then the model rats were treated with point thread embedding.Morris maze test was used to evaluate the learning and memory of the rats.In vivo recording of long term potentiation(LTP) was performed in the dentate gyrus of hippocampus and changes in each experimental group were observed and compared after high-frequency stimulation(HFS).Results: Compared with models group,the results from the Morris tests in treated group(thread embedding group)enhanced significantly(P