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Outline
To observe the effect of Haipiaoxiao（Cuttlebone）drug-containing serum regulating PI3K/Akt/eNOS signaling pathway on proliferation，apoptosis and angiogenesis of vascular endothelial cells（HUVECs）induced by high glucose.
Twenty SD rats were selected，10 rats were randomly gavaged with Haipiaoxiao（Cuttlebone）water decoction and 10 rats with the same amount of normal saline. The corresponding serum was prepared 2 h after the last administration. HUVECs were cultured in vitro，the optimal high glucose concentration was screened，and the cells were induced for 24 h to establish a high glucose damage model. According to the induction conditions，they were divided into blank Control group，Model group，Haipiaoxiao（Cuttlebone）low group，Haipiaoxiao（Cuttlebone）medium group group，Haipiaoxiao（Cuttlebone）high group，and the corresponding serum was treated for 24 h. The cell proliferation was detected by CCK-8 method. Cell migration ability was evaluated by cell scratch assay. The apoptosis of cells in each group was detected by flow cytometry. Angiogenesis of each group was detected by in vitro tube formation. Real-time fluorescence quantitative polymerase chain reaction（qRT-PCR）was used to determine the mRNA expression of each group. The expressions of phosphatidylinositol 3-kinase（PI3K），p-PI3K，protein kinase B（Akt），p-Akt，endothelial nitric oxide synthase（eNOS）and human tumor suppressor gene（p53）protein were detected by Western blot.
The optimal concentration of high glucose induced endothelial cell injury was 30 mmol/L. Compared with those of the Control group，the cell activity，wound-healing rate，the number of angiogenesis experiment in vitro were decreased，the apoptosis rate and p53 protein expression were increased，the protein expressions of p-PI3K，p-Akt and eNOS were decreased，and mRNA expressions of PI3K，Akt and eNOS were decreased in the Model group. The differences were statistically significant（P<0.05）. Compared with the Model group，Haipiaoxiao（Cuttlebone）low，medium and high dose groups increased the cell survival rate，improved the wound-healing rate，increased the number of angiogenesis experiment，decreased the apoptosis rate and p53 protein expression，increased the protein expressions of p-PI3K，p-Akt and eNOS and the mRNA expressions of PI3K，Akt and eNOS. The differences were statistically significant（P<0.05）.
Haipiaoxiao（Cuttlebone）drug-containing serum can mediate PI3K/Akt/eNOS signaling pathway to promote the proliferation，migration and angiogenesis of vascular endothelial cells.
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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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