Full
Outline
To prepare exosomes（Exos）derived from human umbilical cord mesenchymal stem cells（HucMSC）loaded with Forsythoside A（FTA）, and investigate their mechanism of action on acute lung injury（ALI）based on the sphingosine-1-phosphate（S1P）/phosphatidylinositol 3-kinase（PI3K）/protein kinase B1（AKT1）signaling pathway.
HucMSC-Exos were extracted, FTA-Exos were prepared, and characterized through Malvern particle size analyzer, transmission electron microscope and Western blot（WB）experiments. The drug loading of FTA-Exos was determined by high-performance liquid chromatography. An ALI cell model was constructed by stimulating human bronchial epithelial cells（BEAS-2B）with lipopolysaccharide（LPS）, and the cells were divided into a control group, a model group, an FTA group and an FTA-Exos group. The uptake effect of FTA-Exos by BEAS-2B cells was observed using laser confocal microscopy. Flow cytometry was used to detect the level of reactive oxygen species（ROS）in cells from each group. WB experiments were performed to detect the expression levels of S1P, PI3K and AKT1 proteins in cells of each group.
The particle size and morphology of HucMSC-Exos and FTA-Exos both conformed to the biological characteristics of exosomes, and they positively expressed the marker proteins heat shock protein 70（HSP70）, tumor susceptibility gene 101（TSG101）and cluster of differentiation 81（CD81）. The drug loading of FTA-Exos was（8.11 ± 0.36）%. FTA-Exos could be effectively taken up by BEAS-2B cells, and the fluorescence intensity was significantly enhanced at 4 hours. Compared with those in the control group, the levels of ROS and the expressions of S1P, PI3K and AKT1 proteins were significantly increased in the ALI cell model group（P < 0.01）. In comparison with those in the model group, the levels of ROS and the expressions of S1P, PI3K and AKt1 proteins were significantly decreased in both the FTA group and the FTA-Exos group（P < 0.05, P < 0.01）.
The study successfully prepared FTA-Exos and demonstrated that they can regulate ALI cell models by affecting ROS levels and the S1P/PI3K/AKT1 signaling pathway, providing new ideas for the development and application of FTA in the treatment of ALI.
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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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