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Outline
To explore whether Sagittaria Sagittifolia polysaccharide（SSP）plays a protective role in L02 hepatocyte injured by six heavy metals mixture bymediating the Toll-like receptors 4（TLR4）/myeloid differentiation factor 88（MyD88）/nuclear transcription factor-κB（NF-κB）signaling pathway.
Firstly，according to the dietary intake of six heavy metals in rice sold in Beijing，a heavy metal ixture was prepared for inducing L02 cell injury. MTT assay was used to detect the effect of SSP（0.25 mg/mL，0.5 mg/mL，1 mg/mL，2 mg/mL）on the L02 cells survival rate with 24 h pre-treatment. Annexinv FITC/propidium iodide（PI）double staining combined with flow cytometry and Hoechst 33342 staining were used for detecting the cell apoptosis. The content of reactive oxygen species（ROS）in cells was detected by using DCFH-DA fluorescence probe. The protein expressions of poly-ADP ribose polymerase1（PARP1），dynamic related protein 1（Drp1），tumor necrosis factor-α（TNF-α），interleukin-1β（IL-1β），interleukin-17（IL-17），TLR4，MyD88 and NF-κB p65 were detected by Western blot.
MTT assay showed that SSP（1 mg/mL，24 h）had the best protective effect. Compared with those of the model group，the ROS contentand apoptosis rate in SSP treatmentgroup were significantly decreased（P<0.05），and the protein expressions of Drp 1，TNF-α，IL-1β，IL-17，TLR4，MyD88 and NF-κB p65 decreased significantly（P<0.05）and the protein expression of PARP1 increased significantly（P<0.01）.
SSP can effectively improve the apoptosis of L02 cells induced by six heavy metals mixture，whichmay be related to the regulation of TLR4/MyD88/NF-κB signaling pathway to inhibit the oxidative damage and inflammatory response.
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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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