Objective To study protective effects of total polyphenols from Nymphaea candida (NCTP) on lipopolysaccharide (LPS) induced inflammatory injury in A549 cells. Methods Polyphenol content in NCTP was determined by Folin-Ciocalte method, and its characteristic components were detected by high performance liquid chromatography (HPLC). A549 cells were treated with 1.0 μg/mL LPS for 12 h to induce inflammatory damage model, and cell counting kit-8 (CCK-8) method was used to screen out the optimal intervention concentration of NCTP. The experiment was divided into control group, lipopolysaccharide model group, dexamethasone (DXM 0.039 μg/mL) group, and NCTP (6.0, 15.0, 30.0 μg/mL) groups. The levels of nitric oxide (NO), prostaglandin E2 (PGE2), malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione (GSH) were detected by biochemical method. Enzyme-linked immunosorbent assay (ELISA) was used to detect the expression of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in A549 cells. The expression of related proteins [Kelch-like ECH-associating protein 1 (Keap1), nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1)] in nuclear factor erythroid-2-related factor 2/heme oxygenase 1 (Nrf2/HO-1) signaling pathway was detected by Western Blot. Results The content of total polyphenols in NCTP was (90.15±0.17)%, and the content of isostrictiniin, ellagic acid and nicotiflorin was (28.05±0.02)%, (2.15±0.03)% and (3.50±0.01)%, respectively. The mass concentrations of NCTP screened by CCK-8 were 6.0, 15.0, 30.0 μg/mL. After LPS stimulation of A549 cells, the expressions of intracellular inflammatory mediators NO, PGE2 and inflammatory factors IL-1β, IL-6 and TNF-α were significantly increased (P<0.01), indicating that the inflammatory damage model of A549 cells was successfully constructed. Compared with the model group, NCTP significantly inhibited the expression of NO, PGE2, IL-1β, IL-6, TNF-α and MDA and increased the activities of SOD and GSH in LPS-induced A549 cells. Compared with the model group, NCTP also significantly increased the expression of Nrf2 and HO-1 proteins in the damaged cells, and decreased the expression level of Keap1 proteins. Conclusion NCTP has the better protective effect on LPS-induced inflammatory injury in A549 cells, and its mechanism is relative to the regulation of the Nrf2/HO-1 signal pathway. The results of this study can provide a basis for the research and development of functional foods with this effective part to prevent respiratory diseases.
| 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 |