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
10.13422/j.cnki.syfjx.20212409
- VernacularTitle:人参-三七-川芎提取物延缓过氧化氢诱导的内皮细胞衰老中线粒体氧化应激的作用
- Author:
Ye WU
1
;
Qiang WANG
2
;
Cheng-kui XIU
1
;
Yan-hong HU
1
;
Yan-yan MA
1
;
Ying-kun FU
3
;
Xue WANG
1
;
Yan LEI
1
;
Jing YANG
1
Author Information
1. Beijing Key Laboratory of Basic Research on Prevention and Treatment of Major Diseases by Traditional Chinese Medicine，Experimental Research Center，China Academy of Chinese Medical Sciences，Beijing 100700，China
2. Wangjing Hospital，China Academy of Chinese Medical Sciences，Beijing 100102，China
3. Guang'anmen Hospital，China Academy of Chinese Medical Sciences，Beijing 100056，China
- Publication Type:Research Article
- Keywords:
aging;
vascular aging;
aging of endothelial cells;
Ginseng Radix et Rhizoma， Notoginseng Radix et Rhizoma， and Chuanxiong Rhizoma extract;
oxidative stress
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2021;27(24):17-24
- CountryChina
- Language:Chinese
-
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 （H2O2）-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）， H2O2 model group （H2O2）， H2O2 + DMSO group （DMSO， 1 mL·L-1）， resveratrol group （Resv， 8 μmol·L-1）， and low- （200 mg·L-1）， medium- （300 mg·L-1）， and high-dose （400 mg·L-1） GNC （GNC-L， GNC-M， and GNC-H） groups. Except control group and H2O2 group， the other groups were intervened with corresponding agents. Subsequently， 300 μmol·L-1 H2O2 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-β-galactosidase （SA-β-Gal） staining， flow 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-β-gal staining results showed that H2O2 group had increased blue-stained cells compared with other groups （P<0.01）. Compared with those in the control group， the ratio of G0/G1 phase cells significantly increased （P<0.05） and that of G2/M phase cells decreased （P<0.05） in the H2O2 group. Compared with those in the H2O2 group， the proportion of G0/G1 cells decreased （P<0.05） while that of G2/M cells increased （P<0.05） in GNC-H groups and Resv group. The fluorescence staining for determining mitochondrial ROS level showed that the H2O2 group had weakened fluorescence intensity than the control， GNC-H， and GNC-M groups （P<0.05）. The mitochondrial membrane potential fluorescence intensity of the H2O2 group was weaker than that of the control， GNC-H， GNC-M， and GNC-L groups （P<0.01）， as well as the Resv group （P<0.05）. Western blot showed that the protein level of MnSOD was significantly lower in the H2O2 group than in the control， GNS-H， and GNS-M groups （P<0.05）， whereas the protein level of p-p66 showed an opposite trend （P<0.01）， indicating that the medication can alleviate the intracellular mitochondrial oxidative stress. Conclusion:GNC can delay the H2O2-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.
