ABSTRACT
Aim To investigate the possible mechanism of paeonol inhibiting the inflammatory response of fibroblast synovial cells (RA-FLSS) in rheumatoid arthritis. Methods CCK-8 assay was used to detect Paeonol's inhibitory level on the abnormal proliferation of arthritis human fibroblast synovial cells (RA-FLSs). The levels of endoplasmic reticulum stress-related proteins MANF and ATF6 were detected by Western blot. Cell localization of transcription factor p65 and Mesencephalic Astrocyte Derived Neurotrophic Factor (MANF) was detected by immunofluorescence. RT-qPCR detected the changes of p65 target genes. Results Paeonol could significantly inhibit the abnormal proliferation of RA-FLSS cells. Paeonol activates ATF6 and increases the expression of MANF. Paeonol promoted the nuclear transfer of MANF protein and inhibited the transcriptional activity of p65. Conclusion Paeonol promotes the expression of MANF and nuclear transfer through the endoplasmic reticulum stress pathway and affects the progression of RA by inhibiting the transcriptional activity of p65.
ABSTRACT
Objective::To study the effects of Ermiaosan on migration, adhesion and invasion of human fibroblast-liked synovial cells(FLS) and explore its mechanism. Method::Using the human FLS as the research object, the nontoxic concentration of FLS.FLS was determined by methyl thiazolyl tetrazolium (MTT) colorimetric assay for the follow-up experiment. The transwell migration, adhesion and transwell invasion test were used to detect the migration and adhesion of the different concentration of Ermiaosan on FLS, respectively. The expression of interleukin (IL)-1 beta of FLS supernatant was detected by enzyme linked immunosorbent assay (ELISA). Protein in FLS was extracted and protein expression levels of phosphorylated Janus kinase 1 (p-JAK1), p-signal transducer and activator of transcription (STAT1) and p-STAT6 were detected by Western blot. Result::Compared with control group, tumor necrosis factor(TNF)-α (20 μg·L-1) increased the proliferation, migration, adhesion, invasion and the secretion of IL-1β of FLS (P<0.01). Ermiaosan(0.2, 0.4, 0.8 mg·L-1) had no significant effect on the proliferation of FLS induced by TNF-α for 24 h. Within 24 h, the migration, adhesion, invasion, invasion, and secretion of IL-1β of FLS cells induced by TNF-α were also decreased significantly(P<0.05, P<0.01). Compared with the blank group, TNF-α could induce abnormal elevation of p-JAK1, p-STAT1 and p-STAT6 in FLS (P<0.01), while Ermiaosan of 0.2, 0.4, 0.8 g·L-1 could significantly reduce the expression levels of p-JAK1, p-STAT1 and p-STAT6 (P<0.05, P<0.01). Conclusion::Ermiaosan can inhibit the migration, adhesion and invasion of FLS, and its mechanism may be related to the inhibition of the secretion of IL-1β, the mechanism may be related to JAK/STAT pathway.
ABSTRACT
Aim To explore the effect of resveratrol (Res) on fibroblast synovial cells (FLS) inadjuvant arthritis (AA) rats treated with low concentration of H2O2, and the role of mitochondrial oxidative stress proteins deacetylase 3 (SIRT3) and manganese superoxide dismutase (MnSOD). Methods Twenty SD rats were randomly divided into normal group and model group. The AA model was induced by subcutaneous injection of the complete adjuvant in toes of model group. On 28th day after modeling, the rats were killed and the changes in serum oxidative stress index in two groups were detected. The expression of oxidative stress protein of SIRT3 and MnSOD in synovial tissues of knee joint of AA rats were detected by immunohistochemistry stainning. The effect of different concentrations of H2O2or Res treatment on FLS proliferation was observed by CCK-8 method. The effect of Res on apoptosis and the expression proteins SIRT3 and MnSOD of FLS treated with low H2O2concentration were detected by Hoechst 33258 and Western blot. Results Compared with normal group, the serum of AA rats showed oxidative stress. The expression of SIRT3 and MnSOD increased in synovial tissues of AA model rats. Low concentration of H2O2promoted FLS proliferation, and Res could dose-dependently inhibit proliferation treated with low concentration of H2O2, and increased FLS apoptosis. Moreover, with the increase of Res dose, the expression of SIRT3 and MnSOD protein decreased in FLS treated with low concentration H2O2. Conclusions AA rats are in a state of oxidative stress, low concentration of H2O2can promote FLS growth in AA ras, and Res treatment can inhibit FLS proliferation, promoting FLS apoptosis. The mechanism may be related to the reduction of anti-oxidative stress effect.