Effects of total flavonoids of Dracocephalum moldavica on apoptosis of H9c2 cells induced by OGD/R injury and endoplasmic reticulum stress.
10.19540/j.cnki.cjcmm.20241212.703
- Author:
Tian WANG
1
;
Di-Wei LIU
2
;
Tong-Ye WANG
3
;
Xing-Yu ZHANG
1
;
Jian-Guo XING
2
;
Rui-Fang ZHENG
2
Author Information
1. Xinjiang Medical University Urumqi 830011, China.
2. Xinjiang Institute of Pharmacy Urumqi 830002, China.
3. Shihezi University Shihezi 832000, China.
- Publication Type:Journal Article
- Keywords:
H9c2 cells;
apoptosis;
endoplasmic reticulum stress;
oxygen-glucose deprivation and reoxygenation;
total flavonoids of Dracocephalum moldavica;
tunicamycin
- MeSH:
Animals;
Endoplasmic Reticulum Stress/drug effects*;
Apoptosis/drug effects*;
Rats;
Flavonoids/pharmacology*;
Glucose/metabolism*;
Cell Line;
Lamiaceae/chemistry*;
Drugs, Chinese Herbal/pharmacology*;
Oxygen/metabolism*;
Reperfusion Injury/physiopathology*;
Myocytes, Cardiac/cytology*
- From:
China Journal of Chinese Materia Medica
2025;50(5):1321-1330
- CountryChina
- Language:Chinese
-
Abstract:
This study investigated the effects of total flavonoids of Dracocephalum moldavica(TFDM) on apoptosis in rat H9c2 cells induced by endoplasmic reticulum stress(ERS) established by oxygen-glucose deprivation and reoxygenation(OGD/R) injury and tunicamycin(TM), and explored the potential mechanisms. After successful modeling, the following groups were set in this experiment: control group, model(OGD/R or TM) group, and TFDM low-, medium-, and high-dose groups(12.5, 25, and 50 μg·mL~(-1)). The OGD/R injury model was constructed in vitro. Cell proliferation was assessed using the cell counting kit-8(CCK-8) method. The levels of lactate dehydrogenase(LDH) and creatine kinase MB isoenzyme(CKMB) in the cell supernatant were detected. Western blot was used to assess the expression of ERS-related proteins, including glucose regulatory protein 78(GRP78), C/EBP homologous protein(CHOP), activating transcription factor 6(ATF6), and apoptotic proteins B-cell lymphoma 2(Bcl-2) and Bcl-2-associated X protein(Bax). Apoptosis was detected using the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL) method. In the TM-induced ERS model, Western blot was used to measure the expression of ERS pathway-related proteins GRP78, CHOP, inositol-requiring enzyme 1(IRE1), X-box binding protein 1(XBP1), protein kinase RNA-like endoplasmic reticulum kinase(PERK), eukaryotic initiation factor 2α(eIF2α), ATF6, p-ATF6, and apoptotic proteins Bcl-2, Bax, cysteinyl aspartate specific proteinase-12(caspase-12), and cleaved caspase-12. Gene expression of GRP78, CHOP, PERK, and ATF6 was detected by real-time fluorescence quantitative PCR(RT-qPCR). Apoptosis was again detected using the TUNEL method. The results showed that in the OGD/R model, compared with the control group, the levels of LDH and CKMB in the cell supernatant were significantly increased in the OGD/R group. Compared with the OGD/R group, the levels of LDH and CKMB in the TFDM group were significantly reduced. Western blot results revealed that compared with the control group, the expression of ERS-related proteins and Bax in the OGD/R group was significantly increased, while the expression of Bcl-2 was significantly decreased. Compared with the OGD/R group, the expression of ERS-related proteins and Bax in the TFDM groups was significantly reduced, and the expression of Bcl-2 was significantly increased. TUNEL assay showed that apoptosis was significantly decreased after TFDM treatment. In the TM-induced ERS experiment, compared with the control group, the expression of ERS-related genes, ERS-related proteins, and apoptotic proteins in the TM group was significantly increased, while the expression of Bcl-2 was significantly decreased. Compared with the TM group, the expression of ERS-related genes, ERS-related proteins, and apoptotic proteins in the TFDM group was significantly reduced, and the expression of Bcl-2 was significantly increased. These results suggest that ERS exists in the OGD/R-injured H9c2 cell model, and TFDM can effectively inhibit ERS-induced apoptosis. The mechanism may be related to the downregulation of ERS pathway-related proteins and apoptotic proteins.
