Protective Effect and Mechanism of Aqueous Extract of Perillae Folium on Adriamycin-induced Oxidative Injury in HK-2 Cell
10.13422/j.cnki.syfjx.20191238
- VernacularTitle: 紫苏叶水提物对阿霉素致HK-2细胞氧化损伤的保护及机制
- Author:
Yi-ye ZHU
1
;
En-chao ZHOU
1
;
Kun GAO
1
;
Guo-shun HUANG
1
;
Wei LI
1
;
Ping XIA
1
Author Information
1. Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210029, China
- Publication Type:Research Article
- Keywords:
aqueous extract of Perillae Folium;
renal tubular epithelial cells;
adriamycin;
oxidative injury;
mitochondrial apoptosis;
mitogen-activated protein kinase (MAPK) signaling pathway
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2019;25(12):50-57
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate the protective effect of Perillae Folium with aqueous extract (PFAE) on some key factors of Adriamycin (ADR)-induced oxidative injury in human renal tubular epithelial cells(HK-2), including the survival rate, oxidative injury indexes and cell apoptosis,in order to define the underlying mechanism. Method: A model of ADR-induced HK-2 cells oxidative injury was established in vitro, then cell viability was detected by cell counting kit-8 (CCK-8) after intervention with positive reference N-acetylcysteine (NAC) or PFAE (5,15,45 g·L-1) at different concentrations. According to the morphological changes under microscopy, the optimum concentration of PFAE was screened out for the follow-up experiments. Then, the experiments were divided into six groups:blank group, ADR (0.05 g·L-1) group, PFAE (15 g·L-1) group, ADR+PFAE (0.05+15) g·L-1 group, NAC (0.81 g·L-1) group, and ADR+NAC (0.05+81) g·L-1 group. After that, malondialdehyde (MDA), superoxide dismutase (SOD), total antioxidant capacity(TAC) were measured in the cell homogenate after 24 h administration. The level of reactive oxygen species (ROS) was detected by 2',7'-dichloroflurescin diacetate (DCFH-DA) fluorescence probe. Flow cytometry and TdT-mediated dUTP Nick-End Labeling (TUNEL) were used to monitor the cell apoptosis. Western blot was used to observed the expressions of mitochondrial apoptosis-associated proteins, like B lymphocyte tumor-2 gene (Bcl-2), Bcl-2 related X protein (Bax), cysteine aspartate protease-9 (Caspase-9), cysteine aspartate protease-3 (Caspase-3) and poly ADP-ribose polymerase (PARP), as well as their shear bodies. In addition, the phosphorylation protein expressions of p38 mitogen-activated protein kinase (p38 MAPK), extracellular signal-regulated kinase (ERK), c-Jun amino-terminal kinase (JNK) in mitogen-activated protein kinase (MAPK) signaling transduction pathway were detected by Western blot. Result: Compared with blank group, ADR group showed a decreased cell viability (P<0.01), and lower SOD level (P<0.01), but higher expressions of MDA and ROS (P<0.01), and an increased apoptotic rate (P<0.01). The ADR group also increased in rate of Bax/Bcl-2, cleaved Caspase-9/Caspase-9, cleaved Caspase-3/Caspase-3, and cleaved PARP/PARP (P<0.01), as well as the phosphorylation protein expressions of p38 MAPK, ERK and JNK (P<0.05,P<0.01). Compared with the ADR group, both ADR+PFAE groups and ADR+NAC group had higher cell proliferation rates (P<0.01). In addition, the protective effect of PFAE on cells was the most obvious at the concentration of 15 g·L-1. The ATC and SOD levels were increased in ADR+PFAE group and ADR+NAC group (P<0.01), while their content of MDA and ROS, cell apoptosis, relative ratio of apoptotic protein expression, and phosphorylation protein expressions of p38 MAPK and ERK were all decreased (P<0.01). However, there was no effect on the expression of phosphorylated JNK protein. Conclusion: PFAE could alleviate the oxidative injury of HK-2 cells induced by ADR, and have an antioxidant effect, which inhibited cell apoptosis through mitochondrial apoptotic pathway and ERK/p38 MAPK signaling pathway.
