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Outline
To clarify the effects of betulinic acid on autophagic flux and apoptosis in RPMI-8226 multiple myeloma cells, and to explore the underlying mechanisms.
The apoptosis rate and morphological changes were detected by TUNEL and Hoechst 33258 staining methods; the activities of Caspase-3 and Caspase-8 were measured using the Caspase activity assay kits. The expression of various indicator proteins in cells was detected by Western blotting.
Betulinic acid induced apoptosis and morphological changes in RPMI-8226 cells. Z-VAD-FMK inhibited the pro-apoptotic effect of betulinic acid, while Z-DEVD-FMK further enhanced the pro-apoptotic effect of betulinic acid. Additionally, betulinic acid increased the accumulation of autophagy-related proteins LC3B and P62, promote the activation and protein expression of Caspase-8, and subsequently increase the release of AIF. Z-VAD-FMK antagonized the effects of betulinic acid on LC3B, P62, Caspase-8 and AIF, whereas Z-DEVD-FMK synergized with betulinic acid to enhance its effects on LC3B, P62, Caspase-8 and AIF.
Betulinic acid can directly activate Caspase-3 in RPMI-8226 cells. After acting on RPMI-8226 cells, it can also activate Caspase-8 by blocking autophagic flux, increase mitochondrial membrane permeability, and thereby enhance the release of AIF, thereby mediating an apoptotic process independent of Caspase-3 activation.
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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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