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To observe the effects of LPS(lipopolysaccharide)-HDAC3(induced histone deacetylase 3) on the expression of HMGB1(high mobility histone B1) and nuclear translocation in RAW264.7 cells, and the intervention effect of SFI(Shenfu injection). RAW264.7 cells were induced by LPS to establish a cellular inflammatory injury model, and the cells were intervened with SFI at doses of 3, 6, and 12 μL/mL for 24 h. RT-qPCR(real-time fluorescence PCR) was used to detect the transcriptional levels of HDAC3, HMGB1, IL-1β, and TNF-α in the cells, and Western-blot was used to detect the protein expression of HMGB1 and HDAC3, and immune immunoassay to detect the protein expression of HMGB1 and HDAC3.HDAC3 protein expression, immunofluorescence to observe the effect of SFI on the subcellular localization of HMGB1. ELISA to detect the secretion levels of HMGB1, IL-1β, and TNF-α in the cell supernatant. And small interfering RNA(siRNA) after targeting to silence the HDAC3 in RAW264.7 cells. to observe the effect of SFI on HMGB1 subcellular localization. Compared with the control group, the transcription and expression of HDAC3 in RAW264.7 cells in the model group were significantly reduced (P<0.01), and the expression of HMGB1 was significantly elevated (P<0.01) and simultaneously migrated from the nucleus to the cytoplasm. The inflammatory factors in the supernatant of the cells, such as HMGB1, IL-1β and TNF-α, were significantly elevated (P<0.01). And compared with the model group, SFI (6, 12 μL/mL dose group) up-regulated the transcription and expression levels of HDAC3, down-regulated the transcription, expression, and nuclear translocation of the inflammatory factor HMGB1, and inhibited the secretion of HMGB1, IL-1β, and TNF-α in RAW264.7 cells. After targeted silencing of HDAC3, a large amount of HMGB1 was localized in the cytoplasm, and there was no significant change in protein localization after LPS stimulation, and SFI could not reverse the abnormal localization of HMGB1. SFI may inhibit LPS-induced extra-nuclear migration of HMGB1 in RAW264.7 cells by up-regulating HDAC3 expression, which in turn inhibited its downstream 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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