Establishment and characteristic analysis of fibroblast-like synoviocytes in rats with adjuvant arthritis / 中国药理学通报

ABSTRACT

Aims To establish the methods of primary culture of fibroblast-like synoviocytes in rats with adju-vant arthritis(AA-FLS)and analyze the feature and to investigate the possibility of AA-FLS as the model for the RA in vitro.Methods The synovial cells obtained from the SD rats were immunized by the Mtb and iden-tified by morphology and immunocytochemistry.The viability of AA-FLS was assessed by Cell Counting Kit-8.ELISA was applied to detect TNF-αand IL-1 βin cell media. Apoptosis was measured by Hochest 33258.The expressions of mitochondrial apoptosis-re-lated molecules,including Bcl-2,Bax,Pro-caspase-3 and Cleave-caspase-3 were determined by Western Blot.Result In isolated primary synovial cells,more than 95% of AA-FLSs were fusiform.Immunocyto-chemistry result showed a positive expression of vimen-tin and a negative expression of CD68 in AA-FLS.Cell proliferation of AA-FLS was higher than FLS and cell apoptosis of AA-FLS was curbed.Western blot data demonstrated that the protein expressions of Bcl-2,Bax were regulated and the expression of caspase-3 was ac-tivated in AA-FLS.Conclusions AA-FLS is biologi-cally characterized by high level proliferation activity and inflammatory cytokines and apoptosis suppression. AA-FLS can be used as the model for the RA in vitro.