2022, Vol. 26 ›› Issue (31): 4982-4987

Bax/Bcl-2 protein and apoptosis during adipose-derived stromal cell differentiation into astrocytes

Zhang Lili, Ou Ya, Yuan Xiaodong, Zhang Pingshu   

Abstract: BACKGROUND: Neurodegenerative diseases are a large class of major neurological diseases that can lead to the loss of patients’ ability to work and live. At present, there is no effective treatment for this disease. Adipose-derived stromal cells can induce differentiation into nerve cells in vitro, which is expected to provide a new method for the treatment of this kind of diseases.
OBJECTIVE: To explore the expression changes of Bax/Bcl-2 protein during the differentiation of adipose-derived stromal cells into astrocytes, and to further investigate whether there is mitochondrial pathway-mediated apoptosis during the induction.
METHODS: Immunocytochemistry and western blot assay were used to analyze the expression of glial fibrillary acidic protein, Bax, Bcl-2, cytochrome C, and casepase-3 in the process of adipose-derived stromal cell differentiation into astrocytes when induction for 48 hours and 7, 14, and 21 days. Transmission electron microscopy was used to observe the ultrastructure of cells and mitochondria.
RESULTS AND CONCLUSION: (1) After being induced for 48 hours and 7, 14, and 21 days, the expression of glial fibrillary acidic protein reached the peak on day 7 (P < 0.05). There was no significant difference in glial fibrillary acidic protein expression between 14 days and 7 days after induction (P > 0.05). (2) The expression of Bax, cytochrome C, and casepase-3 increased gradually with the induction time extending and reached the peak on day 14 (P < 0.05). The expression of Bcl-2 was decreased gradually and reached the lowest level on day 14 after induction (P < 0.05). (3) Typical apoptotic bodies and mitochondrial damage were observed using transmission electron microscopy. (4) Adipose-derived stromal cells were in the best survival and differentiation state at the 7th to 14th day of differentiation. During adipose-derived stromal cell differentiation into astrocytes, the inhibitory effect of Bcl-2 on apoptosis was significantly weakened, and the pro apoptotic effect of Bax was gradually enhanced. Cytochrome C was released into the cytoplasm by damaged mitochondria, which led to apoptosis in mitochondrial pathway.
Key words: adipose-derived stromal cell, induction, differentiation, astrocyte, mitochondria, apoptosis, protein