The study on the resistance of autophagy of fibroblast-like synovial cells to apoptosis induced by methotrexate in rheumatoid arthritis
10.3760/cma.j.issn.1007-7480.2019.02.007
- VernacularTitle:甲氨蝶呤通过诱导自噬促进类风湿关节炎滑膜成纤维细胞凋亡机制的研究
- Author:
Ke XU
1
;
Yongsong CAI
;
Chao LU
;
Xiaoyu REN
;
Yuanzhen CAI
;
Zhi YANG
;
Peng XU
Author Information
1. 西安交通大学附属红会医院关节外科 710054
- Keywords:
Methotrexate;
Arthritis,rheumatoid;
Fibroblast-like synovial cells;
Apoptosis;
Autophagy
- From:
Chinese Journal of Rheumatology
2019;23(2):106-109,后插1-后插2
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clarify the mechanisms that the response of fibroblast-like synovial (FLS) cellsto methotrexate (MTX) in rheumatoid arthritis (RA) and to provide theory basis for the drug treatment of RA.Methods Synovial fibroblasts were isolated from synovial tissue specimens obtained from patients with RA andexposed to MTX.Cell viability was measured using a MTT assay and cell apoptosis was valued by flow cytometry.Western blotting analysis of LC3 and immunocytochemistry were used to analyze the induction of autophagy in RA-FLS after treating with MTX.Transfection of siRNA was used to interfere the expression of Beclin1 to down-regulate the autophagy,cell apoptosis was valued by flow cytometry and western blot analysis was used to test the PARPp85 with or without the presence of MTX.Statistical product and service solutions (SPSS) 18.0 statistical software was used for statistical analysis of all experimental data.Independent sample t test was used according to data distribution status,homogeneity of variance,and normal distribution.GraphPad Prism 5.0 was used to draw statistical graphs.Results MTX induced apoptosis was increased in RA-FLS.MTX stimulated the autophagy response in RA-FLS by inducing autophagosome formation.In RA-FLS,transfection with Beclin1 siRNA inhibited autophagy and increased the susceptibility to MTX,which induced cell death.Conclusion Autophagy of RA-FLS contributes to the resistance to apoptosis induced by methotrexate.
