Effect of Xinfeng Capsules-containing serum on TNF-α-induced apoptosis and inflammation of fibroblast-like synoviocytes in rheumatoid arthritis.
10.19540/j.cnki.cjcmm.20200915.402
- Author:
Jian-Ting WEN
1
;
Jian LIU
2
;
Xin WANG
1
;
Jie WANG
1
Author Information
1. Anhui University of Traditional Chinese Medicine Hefei 230038, China.
2. Department of Rheumatology, First Affiliated Hospital of Anhui University of Traditional Chinese Medicine Hefei 230031, China.
- Publication Type:Journal Article
- Keywords:
Xinfeng Capsules;
apoptosis;
fibroblast-like synoviocytes;
inflammatory response
- MeSH:
Apoptosis;
Arthritis, Rheumatoid/genetics*;
Capsules;
Cells, Cultured;
Drugs, Chinese Herbal;
Fibroblasts;
Humans;
Inflammation;
Synovial Membrane;
Synoviocytes;
Tumor Necrosis Factor-alpha/genetics*
- From:
China Journal of Chinese Materia Medica
2021;46(2):436-443
- CountryChina
- Language:Chinese
-
Abstract:
The aim of this paper was to observe the effect of Xinfeng Capsules(XFC)-containing serum on the apoptosis and inflammation of fibroblast-like synoviocytes(FLS) in rheumatoid arthritis(RA) induced by tumor necrosis factor-α(TNF-α), so as to investigate the mechanism of XFC in the treatment of RA. RA-FLS immortalized cell line was established, and XFC drug-containing serum was prepared. CCK-8, ELISA, RT-qPCR, immunofluorescence and TUNEL were used to observe the effect of XFC-containing serum on RA-FLS apoptosis and inflammatory indexes. CCK-8 results showed that the optimal concentration and time of TNF-α on RA-FLS were 10 ng·mL~(-1) and 48 h, respectively; and the optimal concentration and time of XFC on RA-FLS were 6.48 mg·g~(-1) and 72 h, respectively. The results of ELISA showed that compared with RA-FLS group, the expressions of TNF-α, IL-1β, IL-6, IL-8 in TNF-α+RA-FLS group were significantly increased, while the expressions of IL-4 and IL-10 were significantly decreased(P<0.01); after intervention with XFC-containing serum, the expressions of TNF-α, IL-1β, IL-6, IL-8 were significantly decreased, whereas the expressions of IL-4 and IL-10 were significantly increased(P<0.01). The results of RT-qPCR showed that compared with RA-FLS group, the mRNA expressions of Fas, FasL, caspase-3, caspase-8, Bax, Bcl-X1 in TNF-α+RA-FLS group were significantly decreased, while the mRNA expression of Bcl-2 was significantly increased(P<0.001); after intervention with XFC-containing serum, the mRNA expressions of Fas, FasL, caspase-3, caspase-8, Bax, Bcl-X1 were significantly increased, whereas the mRNA expression of Bcl-2 was significantly decreased(P<0.01). The results of immunofluorescence showed that compared with RA-FLS group, the protein expressions of caspase-3 and Bax in TNF-α+RA-FLS group was significantly lower than those in RA-FLS group(P<0.05); after intervention with XFC-containing serum, the protein expressions of caspase-3 and Bax were significantly increased, whereas the protein expression of Bcl-2 was significantly decreased(P<0.05). TUNEL results showed that compared with RA-FLS group, the apoptosis of TNF-α+RA-FLS group was decreased(P<0.05); after intervention with XFC-containing serum, the apoptosis was significantly increased(P<0.05). One of the mechanisms of XFC in the treatment of RA is to promote the apoptosis of RA-FLS and inhibit its inflammatory reaction.
