Neuroprotective effect and mechanism of Ginkgo biloba extract on rotenone-induced damage in PC12 cells
10.3760/cma.j.cn371468-20230816-00058
- VernacularTitle:银杏叶提取物对鱼藤酮所致PC12细胞损伤的保护作用及机制研究
- Author:
Fangzhen SHAN
1
;
Nannan ZHANG
Author Information
1. 济宁医学院附属医院医学研究中心，济宁 272029
- Keywords:
Ginkgo biloba extract;
Rotenone;
PC12 cells;
Parkinson disease;
Mitochondrial damage;
Apoptosis
- From:
Chinese Journal of Behavioral Medicine and Brain Science
2024;33(5):445-451
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the protective effects and mechanisms of Ginkgo biloba extract (EGb) on rotenone-induced damage in PC12 cells, a pheochromocytoma cell line derived from rat adrenal medulla.Methods:PC12 cells were divided into blank control group, rotenone model group (0.5 μmol/L rotenone) and EGb761 group (0.5 μmol/L rotenone+ 100 mg/L EGb761). Mitochondrial membrane potential was detected by JC-1 probe. Mitochondrial energy metabolism levels were measured using a Seahorse XFe24 analyzer. Furthermore, protein expression levels of caspase-3, cleaved-caspase-3, adenosine monophosphate-activated protein kinase(AMPK), phosphorylated-AMPK (p-AMPK), phosphatidylinositol-3-kinase(PI3K), phosphorylated-PI3K(p-PI3K), serine/threonine-protein kinase (AKT), phosphorylated-AKT(p-AKT), mammalian target of rapamycin (mTOR), and phosphorylated-mTOR (p-mTOR) were determined by Western blot.SPSS 20.0 software was used for data analysis.One-way ANOVA was used for multiple group comparisons and LSD test was used for further pairwise comparisons.Results:(1) JC-1 staining showed statistically significant differences in the mitochondrial membrane potential of PC12 cells among the three groups ( F=34.89, P<0.05). EGb761 significantly increased the mitochondrial membrane potential in PC12 cells compared to the rotenone model group (fluorescence intensity ratio: (1.30±0.16), (0.81±0.15), P<0.05). (2) Seahorse experiments showed statistically significant differences in the basal respiration, ATP production, maximal respiration and spare respiratory capacity of PC12 cells among the three groups ( F=38.07, 64.18, 64.42, 34.62, all P<0.05). EGb761 significantly increased the basal respiration ((73.02±13.81)pmol/min, (33.69±3.91)pmol/min), ATP production ((57.08±7.31)pmol/min, (18.08±2.50)pmol/min), maximal respiration ((177.70±17.14)pmol/min, (113.12±13.24)pmol/min), and spare respiratory capacity ((104.72±8.58)pmol/min, (79.43±9.35)pmol/min) of PC12 cells compared to the rotenone model group (all P<0.05). (3) Western blot showed statistically significant differences in the ratio of cleaved-caspase-3/caspase-3、p-AMPK/AMPK、p-PI3K/PI3K、p-AKT/AKT、p-mTOR/mTOR in PC12 cells among the three groups ( F=48.05, 28.68, 33.73, 45.81, 17.39, all P<0.05). EGb761 significantly decreased the ratio of cleaved-caspase-3/caspase-3 ((1.95±0.36), (3.56±0.37)) and p-AMPK/AMPK((1.49±0.19), (2.25±0.27)) and increased the ratio of p-PI3K/PI3K((0.75±0.07), (0.44±0.07)), p-AKT/AKT((0.74±0.06), (0.36±0.09)), and p-mTOR/mTOR((0.90±0.06), (0.62±0.07)) in PC12 cells compared to the rotenone model group (all P<0.05). Conclusion:EGb761 has a protective effect on rotenone-induced PC12 cells, which was associated with the inhibition of AMPK activation and the PI3K/AKT/mTOR pathway, increasing mitochondrial activity and inhibiting apoptosis.
