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To investigate the intervention effect and mechanism of β-ecdyssterone on the osteogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs) based on the regulation of autophagy by the adenosine 5′-monophosphate kinase-activated protein (AMPK)/mammalian target of rapamycin (mTOR).
BMSCs cells were divided into control group (normal culture), low-, middle- and high-dose groups (intervened with 0.01, 0.10 and 1.00 μmol·L-1 ecdysterone). The expression of cell genes was determined by real-time quantitative polymerase chain reaction (qRT-PCR); and the relative expression level of cell proteins was determined by immunofluorescence (IF).
The relative expression levels of alkaline phosphatase (ALP) mRNA in the control group, low-, middle and high-dose groups were 1.01±0.14, 1.22±0.05, 1.28±0.05, 1.59±0.20; the relative protein expression levels of Runt-related transcription factor 2 (RUNX2) mRNA were 1.00±0.07, 1.45±0.07, 2.11±0.32, 4.67±1.45; the relative protein expression levels of phosphorylated AMPK protein were 0.07±0.01, 0.11±0.01, 0.06±0.01, 0.18±0.01; and the relative protein expression levels of Sequestosome 1 (p62/SQSTM1) protein were 1.72±0.02、1.67±0.02、0.94±0.01、0.04±0.01; the relative protein expression levels of p-mTOR protein were 0.66±0.01, 0.40±0.01, 0.42±0.01, 0.04±0.01; and the relative protein expression levels of microtubule-associated protein 1 light chain 3 beta (LC3B) protein were 0.07±0.01, 0.20±0.01, 0.87±0.05, 1.27±0.04, respectively. There were statistically significant differences between the low-, middle-, and high-dose groups and the control group (P<0.05, P<0.001).
β-ecdyssterone can regulate autophagy and promote osteogenic differentiation of BMSCs by activating the AMPK/mTOR pathway.
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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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