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
Aim: To investigate the role of manganese superoxide dismutase (MS0D) protein expression in the proliferation and apoptosis of fibroblast-like synoviocyte (RA-FLS) in rheumatoid arthritis mediated by resveratrol(Res). Methods: After RA-FLSs was treated with Res, the expression of MnSOD protein was detected by CCK-8 method, and the expression of Mn- SOD protein was detected by, Western blot. After lentivirus infection with RA-FLSs, 8 mg · L-1 purine mycin was used to screen the infected cells, and three stable cell lines were established; MnSOD overexpression, MnSOD interference and MnSOD control. The mitochondria reactive oxygen species (mtROS) levels of each cell line treated with 5 μmol · L-1 hydrogen peroxide (H2O2) and 200 mol · L-1 Res were detected by laser confocal microscopy, and cell apoptosis was detected by flow cytometry. Results Res could inhibit the proliferation and the expression of MnSOD in RA- FLSs. The lentivirus-mediated MnSOD regulation could significantly increase or decrease the expression of Mn- SOD in RA-FLSs. MnSOD over expression could decrease the level of mtROS and apoptosis of the RA- FLSs treated with 5 μmol · L-1 H2O2 and 200 μmol · L-1 Res. While the MnSOD RNAi showed the opposite results. Conclusions: Res may up-regulate mtROS levels by inhibiting the expression of MnSOD, thus promoting the apoptosis of RA-FLSs.