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To investigate the molecular mechanism by which Astragal side IV (ASV) combined with Quercetin (QUE) inhibits silica (SiO2)-induced macrophage proptosis activation.
An in vitro co-culture system was established using rat alveolar macrophages (NR8383) and rat fibroblasts (RFL-6). The experiment was divided into six groups: control group, drug group, SiO2 group, SiO2 + drug group, SiO2 + Balmacaan (Bel) group, and SiO2 + drug + Bel group. After 24 hours of intervention, cell and supernatant samples were collected. The levels of transforming growth factor β1 (TGF-β1), tumor necrosis factor alpha (TNF-α), and high mobility group box-1 (HMGB1) in the supernatant were measured using ELISA. The protein levels of α-SMA,Vimentin, and E-Cadherin (E-cad) in RFL-6 cells were detected using Western blot. The mRNA and protein expression levels of Caspase-1, interleukin-1β (IL-1β), and IL-18 in NR8383 cells were assessed using PCR and Western blot, respectively.
Compared to the control group, the levels of TGF-β1, TNF-α, and HMGB1 in the supernatant of the SiO2 group were significantly increased (P<0.05), the protein levels of α-SMA and Vimentin in RFL-6 cells were elevated, while E-cad protein levels were reduced. The expression levels of (Cleaved-) Caspase-1, IL-1β, and IL-18 mRNA and protein in NR8383 cells were also significantly increased (P<0.05). Compared to the SiO2 group, the levels of TGF-β1, TNF-α, and HMGB1 in the supernatants of the SiO2 + drug group, SiO2 + Bel group, and SiO2 + drug + Bel group were significantly decreased (P<0.05). The protein levels of α-SMA and Vimentin in RFL-6 cells were reduced, and E-cad protein levels were elevated. The expression levels of (Cleaved-) Caspase-1, IL-1β, and IL-18 mRNA and protein in NR8383 cells were also significantly decreased (P<0.05).
ASV combined with QUE may reduce the secretion of inflammatory factors by inhibiting the proptosis levels of macrophages exposed to SiO2 in the co-culture system, thereby suppressing the fibrosis levels in fibroblasts.
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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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