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Effect of Stemona tuberosa Alkaloids on Apoptosis of SMMC-7721 Cells and Expression of Bcl-2, Bax, and Cleaved Caspase-3 / 中国实验方剂学杂志
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 73-79, 2021.
Article in Chinese | WPRIM | ID: wpr-906052
ABSTRACT

Objective:

To investigate the effect of<italic> Stemona tuberosa</italic> alkaloids on the apoptosis of human hepatoma SMMC-7721 cells and the expression of apoptosis-related proteins including B lymphocytoma-2 (Bcl-2), Bcl-2-associated X protein (Bax), and cleaved cysteinyl aspartate-specific protease-3 (cleaved Caspase-3).

Method:

SMMC-7721 cells were routinely cultured, passaged, and treated with various concentrations (50, 75, 112, 167, and 250 mg·L<sup>-1</sup>) of <italic>S. tuberosa </italic>alkaloids, while those in the blank control group were only treated with 10% fetal bovine serum. The cell proliferation was determined by tetrazolium bromide (MTT) colorimetry and colony assay and the cell apoptosis by Hoechst 33258 staining. The protein expression levels of Bcl-2, Bax, and cleaved Caspase-3 were detected by Western blot.

Result:

<italic>S. tuberosa</italic> alkaloids inhibited the proliferation of SMMC-7721 cells, and the inhibition rate was significantly increased in comparison with that in the blank control group (<italic>P</italic><0.01), with the half maximal inhibitory concentrations (IC<sub>50</sub>) at 24 h, 48 h, and 72 h being (173.36±8.75), (112.14±16.50), and (96.41±2.60)mg·L<sup>-1</sup>, respectively. The cell colony-inhibitory activity was significantly increased in a dose-dependent manner (<italic>P</italic><0.01). Compared with the blank control group, <italic>S. tuberosa</italic> alkaloids promoted the apoptosis of SMMC-7721 cells, manifested as increased number of apoptotic cells and elevated apoptotic rate (<italic>P</italic><0.01). The typical morphological changes such as brightly blue-fluorescent condensed nuclei, cytoplasmic shrinking, and karyopyknosis were found under the upright fluorescence microscope. As revealed by comparison with the blank control group, the expression of Bcl-2 was significantly down-regulated (<italic>P</italic><0.01), while the protein expression levels of pro-apoptotic protein Bax and cleaved Caspase-3 in the 75, 112, 167, and 250 mg·L<sup>-1</sup> <italic>S. tuberosa</italic> alkaloids groups were significantly up-regulated (<italic>P</italic><0.01).

Conclusion:

<italic>S. tuberosa </italic>alkaloids inhibit the proliferation of SMMC-7721 cells and promote their apoptosis possibly by inhibiting Bcl-2 protein expression and promoting Bax and cleaved Caspase-3 protein expression.

Full text: Available Index: WPRIM (Western Pacific) Language: Chinese Journal: Chinese Journal of Experimental Traditional Medical Formulae Year: 2021 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Language: Chinese Journal: Chinese Journal of Experimental Traditional Medical Formulae Year: 2021 Type: Article