Cytotoxicity and mitochondrial injury mechanism of matrine on PC12 cells
10.3867/j.issn.1000-3002.2017.09.003
- VernacularTitle:苦参碱对PC12细胞的毒性及线粒体损伤机制
- Author:
Fang SHEN
1
;
Pei LIANG
;
Hong LU
Author Information
1. 浙江中医药大学药学院
- Keywords:
matrine;
neurotoxicity;
PC12 cells;
reactive oxygen species;
mitochondrial damage;
apoptosis
- From:
Chinese Journal of Pharmacology and Toxicology
2017;31(9):873-879
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE To study the toxicological effect of matrine (MT) on PC12 cells and the mechanism of mitochondrial damage. METHODS After treatment with MT 2, 4 and 8 mmol·L-1for 8, 16,24 and 48 h,respectively,the viability of PC12 cells was measured with methylthiazolyltetrazolium assay.PC12 cells were treated with MT 2,4 and 8 mmol·L-1for 24 h,before the morphological changes were observed by Hoechst33342 staining. The superoxide dismutase (SOD) activity and malondialde-hyde (MDA) content in cells were detected using hydroxylamine method and thiobarbituric acid method, apoptosis rate and reactive oxygen species (ROS) of PC12 cells were detected with flow cytometry, mitochondrial membrane potential (MMP) was detected with JC-1 staining, and the expressions of procaspase 3, procaspase 9, cleaved-caspase 3, Bax and Bcl-2 were detected with Western blotting. RESULTS MT inhibited the growth of PC12 cells in a time-and concentration-dependent manner.After being treated with MT for 24 h,the nuclei of PC12 cells in MT groups showed chromatin agglutination and partial rupture to different degrees,and compared with normal control group,the apoptosis rates of MT 2,4 and 8 mmol·L-1groups were significantly increased(P<0.01).Intracellular ROS and MDA levels increased (P<0.05, P<0.01), SOD activity decreased (P<0.01), and MMP decreased. The expressions of procaspase 9, procaspase 3 and Bcl-2 were significantly decreased (P<0.01, P<0.05), the expres-sions of cleaved-caspase 3 and Bax were significantly increased(P<0.05,P<0.01),and the ratio of Bcl-2/Bax was significantly decreased (P<0.01). CONCLUSION MT has toxic effect on PC12 cells and induces apoptosis through ROS mediated mitochondrial pathway.
