Journal of Shanghai Jiao Tong University (Medical Science) >

2021 , Vol. 41 >Issue 11: 1446 - 1453

DOI: https://doi.org/10.3969/j.issn.1674-8115.2021.11.007

Basic research

Effect of PDSORBS2 peptide on H9C2 cell apoptosis induced by hypoxia and reoxygenation

  • Lu-lu CHEN ,
  • Xun-long XU ,
  • Wan-lan CHEN ,
  • Zhao-hui QIU
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  • Department of Cardiology, Tongren Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200336, China
QIU Zhao-hui, E-mail: qzh3503@shtrhospital.com.

Online published: 2021-12-03

Supported by

Key Specialist Project of Cardiovascular Medicine in Changning District, Shanghai(20161001)

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Abstract

Objective

·To explore the inhibitory effect and its mechanism of the PDSORBS2 peptide on H9C2 cell apoptosis induced by hypoxia and reoxygenation (H/R).

Methods

·In the previous work of this study, a new peptide PDSORBS2 (LPASLNS) was discovered from rat cardiomyocytes after ischemia. Cell counting kit-8 (CCK-8) was used to detect cell viability to select the appropriate concentration of PDSORBS2 peptide and the appropriate time for H/R treatment. The cells were randomly divided into normal control group (NC group), NC+PDSORBS2 group, H/R group and H/R+PDSORBS2 group. A fluorescence microscope was used to observe the changes in the nucleus morphology. The degree of apoptosis and cell cycle was analyzed by flow cytometry. A kit was used to detect the content of reactive oxygen species (ROS) in cells. The expression levels of apoptosis-related proteins including poly ADP-ribose polymerase (PARP), cleaved-caspase3, B-cell lymphoma-2 (Bcl-2), and BCL2-associated X (Bax), extracellular regulated protein kinases (ERK), protein kinase B (AKT), cyclin-dependent kinases 2 (CDK2), and p27Kip1 protein were analyzed by Western blotting in H9C2 cells.

Results

·Compared with the NC group, 6 h of hypoxia and 2 h of reoxygenation significantly decreased the viability of H9C2 cells, while 50 μmol/L PDSORBS2 significantly increased the viability of H9C2 cells induced by H/R (P=0.004). Compared with the H/R group, the pretreatment of PDSORBS2 improved the morphology of H9C2 nuclei induced by H/R and reduced the apoptotic rate of H9C2 cells (P=0.000), cell cycle arrest (P=0.000), and intracellular ROS content (P=0.005). The expression levels of pro-apoptotic proteins (PARP, Bax and cleaved-caspase3) and p27Kip1 protein were down-regulated, and the expression levels of Bcl-2, phospho-extracellular regulated protein kinases (P-ERK), phospho-protein kinase B (P-AKT), CDK2 etc. increased.

Conclusion

·The PDSORBS2 may inhibit H/R-induced apoptosis of H9C2 cells through the ERK/AKT/CDK2/p27Kip1 signaling pathway.

Key words： PDSORBS2 peptide; hypoxia and reoxygenation (H/R); apoptosis; H9C2 cells

Cite this article

Lu-lu CHEN , Xun-long XU , Wan-lan CHEN , Zhao-hui QIU . Effect of PDSORBS2 peptide on H9C2 cell apoptosis induced by hypoxia and reoxygenation[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2021 , 41(11) : 1446 -1453 . DOI: 10.3969/j.issn.1674-8115.2021.11.007

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