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Outline
To investigate the inhibitory effect of Xianhecao（Agrimoniae Herba）extract on lung cancer cell A549 via ferroptosis and the mechanism.
The extract of Xianhecao（Agrimoniae Herba）（XHC）was prepared，and the cells were divided into control group（XHC 0 mg/mL），XHC 5 mg/mL group，XHC 10 mg/mL group and XHC 20 mg/mL group. The cell proliferation was detected by CCK-8 assay，and the cell clonogenic ability was detected by clonogenic assay. The levels of malondialdehyde（MDA），glutathione（GSH）and superoxide dismutase（SOD）in four groups of cells were detected by colorimetric assay，and the mRNA expression levels of NOX1，COX2，GPX4，FTH1 and SLCA11 were detected by RT-qPCR. The ferroptosis related proteins were detected by Western blot，and cellular ferrous ion contents were observed by fluorescence.
Compared with 0 mg/mL，different concentrations of XHCinhibited the proliferation ability of A549 cells in a concentration dependent manner. Clonogenic assay results showed that compared with those of the control group，the number and size of cell clumps formed by the clones from A549 cells treated with XHC decreased，and the clonogenic rate decreased（P<0.05）. In A549 cells treated with XHC，the contents of SOD and GSH significantly decreased，while MDA significantly increased（P<0.05）compared with the control cells. The mRNA and protein expression levels of NOX1 and COX2 were significantly increased in the groups treated with XHC，while those of GPX4，FTH1 and SLCA11 were significantly decreased（P<0.05）. Compared with that of the control group，the cellular Fe2＋fluorescence intensity was significantly enhanced in the XHCtreated groups（P<0.05）.
XHC inhibited proliferation and clonogenicity in lung cancer A549 cells in a concentration dependent manner，and the inhibitory mechanism of XHC is likely to be associated with ferroptosis.
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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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