Study on the Effects of Atractylodes macrocephala Ethanol Extract on Life Span of Caenorhabditis elegans and Its Mechanism
- VernacularTitle:白术乙醇提取物对秀丽隐杆线虫寿命的影响及其机制研究
- Author:
Jianping ZHAO
1
,
2
;
Limin WU
1
,
2
;
Xiaona LU
1
,
2
;
Hui WANG
2
;
Pan WANG
1
;
Zhenqiang ZHANG
1
;
Jiangyan XU
1
;
Zhishen XIE
1
Author Information
1. Academy of Chinese Medicine Sciences,Henan University of TCM,Zhengzhou 450046,China
2. Phar macy College,Henan University of TCM,Zhengzhou 450046,China
- Publication Type:Journal Article
- Keywords:
Atractylodes macrocephala;
Ethanol extract;
Caenorhabditis elegans;
Life span;
Oxidant stress;
Nrf2;
SKN-1
- From:
China Pharmacy
2021;32(4):418-424
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE: To study the effects of Atractylodes macrocephala ethanol extract (AM) on life span of Caenorhabditis elegans(called N 2 nematode for short ),and to investigate its mechanism based on transcription factor SKN- 1/ nuclear factor E 2 related factor 2(Nrf2). METHODS :N2 nematode were divided into blank control group ,positive control group (100 μ mol/L curcumin,similarly hereinafter ),AM low-dose ,medium-dose and high-dose groups (100,200,300 μ g/mL, similarly hereinafter ). The effects of AM on the life span (by average survival time )of N 2 nematode under normal condition and oxidant stress condition (40 mmol/L H 2O2)as well as its effects on reproductive capability (by the number of filial generation )of N2 nematode under normal condition were investigated . 700 μmol/L H2O2 was used to establish neuroblastoma cells N 2a oxidant stress model. Effects of positive control ,low-dose,medium-dose and high-dose of AM on the survival rate of model cells were detected by MTT method. After human embryonic renalepithelial cells 293T were transfected with Nrf 2-ARE plasmid , the effects of positive control and AM on luciferase activity of Nrf2-ARE were detected by luciferase reporter gene method at low,medium and high dose for 24 h and at medium dose for 12,18 and 24 h. RT-PCR was used to detect the effects ofpositive control ,low-dose,medium-dose and high-dose of AM on the mRNA expression of downstream genes NQO- 1 and HO- 1 of Nrf 2 in N 2a cells as well as mRN A expression of en@hactcm.edu.cn downstream genes GCS- 1,GST-7,GST-10,HSP-60,HSP- 16.2 and SOD- 3 of SKN- 1 in N 2 nematode. RESULTS :Compared with blank control group ,average survival time of N 2 nematode under normal and oxidant stress condition was significantly prolonged in positive control group and AM groups ;the number of filial generation on the first day (except for AM high-dose group ),the number of filial generation on the second day (except for AM low-dose group ) and the total number of filial generation (except for AM low-dose group ) were increased significantly(P<0.05 or P<0.01). The survival rate of N 2a cells in positive control group ,AM medium-dose and high-dose groups were significantly higher than that of model group (P<0.05 or P<0.01). Compared with blank control group ,Nrf2-ARE luciferase relative activity of 293T cells in positive control group and AM groups as well as Nrf 2-ARE luciferase relative activity of 293T cells in AM medium-dose group after different time of treatment were increased significantly (P<0.01),in dose-dependent and time-dependent trend. Compared with blank control group ,mRNA relative expression of HO- 1 and NQO- 1(except for positive control group ),GCS-1(except for AM low-dose group ),GST-7(except for positive control group and AM low-dose group ), GST-10 and HSP- 60(except for AM low-dose group ),HSP-16.2(except for positive control group and AM low-dose group )and SOD-3 (except for positive control group and AM low-dose group ) were increased significantly (P<0.05 or P<0.01). CONCLUSIONS:AM can prolong the life span of N 2 nematode under normal and oxidant stress condition and improve the its reproductive capacity ,the mechanism of which may be associated with the activation of SKN- 1/Nrf2 signaling pathway.