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Objective To investigate the effect and mechanism of salvianolic acid B (SB) on proliferation and apoptosis of human rheumatoid arthritis (RA) synovial fibroblasts. Methods Human RA synovial fibroblasts MH7A cells were treated with different concentrations of SB (0, 1, 10, 20, 30, 40 and 50 μmol/L). MTT method was used to detect cell proliferation and flow cytometry was used to detect cell apoptosis, and the optimal concentration of SB was selected. MH7A cells were set as control group(normal cultured cells), SB group (cells cultured with 50 μmol/L SB) and SB+3-MA group (cells cultured with 50 μmol/L SB and 5 mmol/L autophagy inhibitor 3-MA). MTT and flow cytometry were used to detect cell proliferation and apoptosis, qRT-PCR was used to measure the relative expression level of caspase-3 mRNA. Western blotting was used to detect the relative expression levels of caspase-3, LC3-Ⅰ, LC3-Ⅱ, Beclin-1 and p62 protein. Results Compared with 0 μmol/L SB group, the proliferation rate decreased obviously (P<0.05) and the apoptosis rate increased significantly (P<0.05) of MH7A cells treated with 30, 40 and 50 μmol/L SB and 50 μmol/L SB was used for follow-up experiments. Compared with control group, the cell proliferation rate and the relative expression level of p62 protein in SB group decreased significantly, while the apoptosis rate, the relative expression levels of caspase-3 mRNA and protein, as well as the relative expression levels of LC3-Ⅱ/LC3-Ⅰ and Beclin-1 protein increased (P<0.05). Compared with SB group, the proliferation rate and the relative expression level of p62 protein increased obviously (63.47%±1.94% vs.51.33%±4.67%; 0.97±0.06 vs. 0.20±0.02) in SB+3-mA group, and the apoptosis rate (21.4%±1.8% vs. 30.4%±0.6%), the relative expression levels of caspase-3 mRNA (0.83±0.06 vs. 1.27±0.15) and protein (0.64±0.05 vs. 1.10±0.09), as well as the relative expression levels of LC3-Ⅱ/LC3-Ⅰ (0.29±0.07 vs. 1.23±0.21) and Beclin-1 protein (0.36±0.05 vs. 1.05±0.08) decreased(P<0.05). Conclusion SB might inhibit the proliferation and promote apoptosis of human RA synovial fibroblast by inducing autophagy.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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