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Effect of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma, and Chuanxiong Rhizoma Extract on Mitochondrial Oxidative Stress in Hydrogen Peroxide-induced Endothelial Cell Aging / 中国实验方剂学杂志
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-905890
Biblioteca responsável: WPRO
ABSTRACT

Objective:

To observe the effect of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma, and Chuanxiong Rhizoma extract (GNC) on mitochondrial oxidative stress in hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>)-induced aging of human umbilical vein endothelial cells (HUVECs), and explore the therapeutic mechanism of GNC on aging HUVECs.

Method:

The HUVECs were classified into the control group (control), H<sub>2</sub>O<sub>2</sub> model group (H<sub>2</sub>O<sub>2</sub>), H<sub>2</sub>O<sub>2</sub> + DMSO group (DMSO, 1 mL·L<sup>-1</sup>), resveratrol group (Resv, 8 μmol·L<sup>-1</sup>), and low- (200 mg·L<sup>-1</sup>), medium- (300 mg·L<sup>-1</sup>), and high-dose (400 mg·L<sup>-1</sup>) GNC (GNC-L, GNC-M, and GNC-H) groups. Except control group and H<sub>2</sub>O<sub>2</sub> group, the other groups were intervened with corresponding agents. Subsequently, 300 μmol·L<sup>-1</sup> H<sub>2</sub>O<sub>2</sub> was given to other groups except the control group for 4 h to induce aging, and then the cells were cultured in normal media for 24 h. The aging degree, cell cycle, and mitochondrial reactive oxygen species (mtROS) level were determined by SA-<italic>β</italic>-galactosidase (SA-<italic>β</italic>-Gal) stainingflow cytometry, and MitoSox red fluorescence staining, respectively. JC-10 was used as a fluorescent probe to detect the changes in mitochondrial membrane potential, and Western blot was performed to detect the expression of manganese superoxide dismutase (MnSOD) and p-p66 proteins.

Result:

The SA-<italic>β</italic>-gal staining results showed that H<sub>2</sub>O<sub>2</sub> group had increased blue-stained cells compared with other groups (<italic>P</italic><0.01). Compared with those in the control group, the ratio of G<sub>0</sub>/G<sub>1</sub> phase cells significantly increased (<italic>P</italic><0.05) and that of G<sub>2</sub>/M phase cells decreased (<italic>P</italic><0.05) in the H<sub>2</sub>O<sub>2</sub> group. Compared with those in the H<sub>2</sub>O<sub>2</sub> group, the proportion of G<sub>0</sub>/G<sub>1</sub> cells decreased (<italic>P</italic><0.05) while that of G<sub>2</sub>/M cells increased (<italic>P</italic><0.05) in GNC-H groups and Resv group. The fluorescence staining for determining mitochondrial ROS level showed that the H<sub>2</sub>O<sub>2</sub> group had weakened fluorescence intensity than the control, GNC-H, and GNC-M groups (<italic>P</italic><0.05). The mitochondrial membrane potential fluorescence intensity of the H<sub>2</sub>O<sub>2</sub> group was weaker than that of the control, GNC-H, GNC-M, and GNC-L groups (<italic>P</italic><0.01), as well as the Resv group (<italic>P</italic><0.05). Western blot showed that the protein level of MnSOD was significantly lower in the H<sub>2</sub>O<sub>2</sub> group than in the control, GNS-H, and GNS-M groups (<italic>P</italic><0.05), whereas the protein level of p-p66 showed an opposite trend (<italic>P</italic><0.01), indicating that the medication can alleviate the intracellular mitochondrial oxidative stress.

Conclusion:

GNC can delay the H<sub>2</sub>O<sub>2</sub>-induced aging of vascular endothelial cells. The GNC intervention significantly regulated the mitochondrial ROS, mitochondrial membrane potential, and related proteins MnSOD and p-p66 to alleviate oxidative stress. Chinese medicinal materials may delay the aging of vascular endothelial cells by inhibiting mitochondrial oxidative stress.

Texto completo: Disponível Base de dados: WPRIM (Pacífico Ocidental) Idioma: Chinês Revista: Chinese Journal of Experimental Traditional Medical Formulae Ano de publicação: 2021 Tipo de documento: Artigo
Texto completo: Disponível Base de dados: WPRIM (Pacífico Ocidental) Idioma: Chinês Revista: Chinese Journal of Experimental Traditional Medical Formulae Ano de publicação: 2021 Tipo de documento: Artigo
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