Mechanism of atractylenolide Ⅲ in alleviating H9c2 cell apoptosis through ROS/GRP78/caspase-12 signaling pathway based on molecular docking.
10.19540/j.cnki.cjcmm.20220304.401
- Author:
Meng-Yu ZUO
1
;
Tong-Juan TANG
2
;
Peng ZHOU
3
;
Xiang WANG
4
;
Rui DING
1
;
Jin-Fan GU
1
;
Jian CHEN
3
;
Liang WANG
3
;
Juan YAO
2
;
Xiang-Yang LI
1
;
Jin-Ling HUANG
3
Author Information
1. School of Chinese Medicine, Anhui University of Chinese Medicine Hefei 230012, China.
2. Institute of Integrated Chinese and Western Medicine, Anhui Academy of Traditional Chinese Medicine Hefei 230012, China.
3. Institute of Integrated Chinese and Western Medicine, Anhui Academy of Traditional Chinese Medicine Hefei 230012, China Anhui Province Key Laboratory of Chinese Medicinal Formula Hefei 230012, China.
4. School of Nursing, Anhui University of Chinese Medicine Hefei 230012, China.
- Publication Type:Journal Article
- Keywords:
apoptosis;
atractylenolide Ⅲ;
endoplasmic reticulum stress;
heart failure;
molecular docking
- MeSH:
Apoptosis;
Calcium/pharmacology*;
Caspase 12/metabolism*;
Caspase 3/metabolism*;
Endoplasmic Reticulum Chaperone BiP;
Endoplasmic Reticulum Stress;
Lactones;
Molecular Docking Simulation;
RNA, Messenger;
Reactive Oxygen Species/metabolism*;
Sesquiterpenes;
Signal Transduction;
Superoxide Dismutase/metabolism*
- From:
China Journal of Chinese Materia Medica
2022;47(16):4436-4445
- CountryChina
- Language:Chinese
-
Abstract:
This study aims to investigate the effect of atractylenolide Ⅲ(ATL-Ⅲ) on hydrogen peroxide(H_2O_2)-induced endoplasmic reticulum stress and apoptosis of H9 c2 cells via the ROS/GRP78/caspase-12 signaling pathway.The binding activity of ATL-Ⅲ to GRP78 was determined by molecular docking.The result showed that ATL-Ⅲ had a good binding activity to GRP78, and the binding activity of ATL-Ⅲ was stronger than that of its specific inhibitor.The endoplasmic reticulum stress model of H9 c2 was established by H_2O_2(100 μmol·L~(-1)) treatment.Five groups were designed: blank control group, model group, and ATL-Ⅲ(15, 30, and 60 μmol·L~(-1)) groups.Apoptosis was detected by Hoechst/PI double staining and flow cytometry.The levels of superoxide dismutase(SOD), malondialdehyde(MDA), and lactate dehydrogenase(LDH) were measured by colorimetry.The levels of reactive oxygen species(ROS) and calcium(Ca~(2+)) in cytoplasm were determined by the fluorescence probe DCFH-DA and the calcium fluorescence probe Flou-4, respectively.The protein levels of GRP78, caspase-12, and caspase-3 were determined by Western blot, and the mRNA levels of GRP78 and caspase-12 by RT-qPCR.N-acetyl-L-cysteine(NAC) and 4-phenylbutyric acid(4-PBA) were respectively used to inhibit ROS and GRP78, and then the mechanism of ATL-Ⅲ in protecting the cells from endoplasmic reticulum stress induced by H_2O_2 were deduced.ATL-Ⅲ(15, 30, and 60 μmol·L~(-1)) decreased the apoptosis rate and ROS, MDA, and LDH levels(P<0.01), increased the SOD activity(P<0.01), and down-regulated the protein levels of GRP78, caspase-12, and caspase-3 and the mRNA levels of GRP78 and caspase-12(P<0.05).The addition of NAC decreased the apoptosis rate and ROS, MDA, GRP78, caspase-12, and caspase-3 levels(P<0.01), while it elevated the SOD level(P<0.01).The addition of 4-PBA also decreased the apoptosis rate and the levels of GRP78, caspase-12, caspase-3, and Ca~(2+)(P<0.01).The effect of inhibitors were consistent with that of ATL-Ⅲ.In conclusion, ATL-Ⅲ can protect H9 c2 cardiomyocytes by regulating ROS/GRP78/caspase-12 signaling pathway to inhibit H_2O_2-induced endoplasmic reticulum stress and apoptosis.
