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Outline
To investigate the protective effect and intervention mechanism of arctigenin on lipopolysaccharide（LPS）combined with adenosine triphosphate（ATP）-stimulated pyroptosis in RAW264.7 cells.
CCK8 assay was used to detect the effects of different concentrations of arctigenin on the viability of RAW264.7 cells；CCK8 assay was used to detect the effects of intervention of arctigenin（20，40，80μmol/L）on the viability of RAW264.7 cells induced by LPS+ATP；LDH release was used to detect the cellular membrane damage；ELISA was used to detect tumour necrosis factor-α（TNF-α），interleukin-6（IL-6）and IL-1βin cell supernatant；Western blotting was used to detect intracellular NLRP3，Caspase-1 p20，Caspase-1 p45，GSDMD-N and GSDMD-FL protein expression；the number of positive cells stained with propidium iodide（PI）was observed by forward and inverted fluorescence microscopy.
Arctigenin concentration less than 160μmol/L had little effect on cell viability；arctigenin（20，40，80μmol/L）intervention reversed the LPS+ATP-induced decrease in cell viability；arctigenin reduced LPS+ATP-induced release of LDH from pyroptosis cells as well as the number of PI-positive cells；arctigenin reduced LPS+ATP-induced LDH release from pyroptosis cells as well as the number of PI-positive cells；arctigenin inhibited the secretion of pro-inflammatory cytokines TNF-α，IL-6 and IL-1βby pyroptosis cells；arctigenin inhibited the expression of NLRP3，Caspase-1 p20，Caspase-1 p45，GSDMD-N and GSDMD-FL proteins in pyroptosis cells that inhibited the activation of NLRP3 inflammasome.
Arctigenin inhibits NLRP3 inflammasome activation to improve pyroptosis and alleviate inflammatory responses.
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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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