Trichosanthis Pericarpium Aqueous Extract Protects H9c2 Cardiomyocytes from Hypoxia/Reoxygenation Injury by Regulating PI3K/Akt/NO Pathway
Abstract
:1. Introduction
2. Results
2.1. TPAE Inhibited Cell Death Induced by H/R
2.2. TPAE Prevented Apoptosis in H9c2 Cells after H/R
2.3. TPAE Activated PI3K/Akt Signaling Pathway
2.4. Inhibition of PI3K Alleviated the Cardioprotection of TPAE
2.5. TPAE Mitigated H/R-Induced Decrease of NO Production via PI3K/Akt/NOS Pathway
3. Discussion
4. Materials and Methods
4.1. Materials and Chemicals
4.2. Extract Preparation
4.3. Cell Culture and Hypoxia/Reoxygenation Model (H/R)
4.4. Experimental Protocols
4.5. Flow Cytometric Detection of Apoptosis
4.6. Enzyme-Linked Immunosorbent Assay
4.7. RNA Isolation and Purification
4.8. Quantitative Real-Time PCR to Confirm Markers
4.9. Western Blotting Analysis
4.10. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Sample Availability: Samples of the compounds are available from the authors. |
Gene Symbol | Reference Sequence | Primer | Sequence(5′-3′) | Product Size (bp) |
---|---|---|---|---|
Akt | NM_033230.2 | Forward | TCACCTCTGAGACCGACACC | 110 |
Reverse | CCGTTCACTGTCCACACACTC | |||
eNOS | NM_021838.2 | Forward | GGGATTCTGGCAAGACCGAT | 151 |
Reverse | TTGTCCAAACACTCCACGCT | |||
iNOS | NM_012611.3 | Forward | TTCCTCAGGCTTGGGTCTTGT | 213 |
Reverse | ATCCTGTGTTGTTGGGCTGG | |||
Bcl-2 | NM_016993.1 | Forward | GGATCCAGGATAACGGAGGC | 141 |
Reverse | ATGCACCCAGAGTGATGCAG | |||
Bax | NM_017059.2 | Forward | GGGCCTTTTTGCTACAGGGT | 106 |
Reverse | TTCTTGGTGGATGCGTCCTG | |||
GAPDH | NM_017008.3 | Forward | GGCACAGTCAAGGCTGAGAATG | 143 |
Reverse | ATGGTGGTGAAGACGCCAGTA |
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Chu, D.; Zhang, Z. Trichosanthis Pericarpium Aqueous Extract Protects H9c2 Cardiomyocytes from Hypoxia/Reoxygenation Injury by Regulating PI3K/Akt/NO Pathway. Molecules 2018, 23, 2409. https://doi.org/10.3390/molecules23102409
Chu D, Zhang Z. Trichosanthis Pericarpium Aqueous Extract Protects H9c2 Cardiomyocytes from Hypoxia/Reoxygenation Injury by Regulating PI3K/Akt/NO Pathway. Molecules. 2018; 23(10):2409. https://doi.org/10.3390/molecules23102409
Chicago/Turabian StyleChu, Donghai, and Zhenqiu Zhang. 2018. "Trichosanthis Pericarpium Aqueous Extract Protects H9c2 Cardiomyocytes from Hypoxia/Reoxygenation Injury by Regulating PI3K/Akt/NO Pathway" Molecules 23, no. 10: 2409. https://doi.org/10.3390/molecules23102409