Clematichinenoside (AR) Attenuates Hypoxia/Reoxygenation-Induced H9c2 Cardiomyocyte Apoptosis via a Mitochondria-Mediated Signaling Pathway.

Abstract

Mitochondria-mediated cardiomyocyte apoptosis is involved in myocardial ischemia/reperfusion (MI/R) injury. Clematichinenoside (AR) is a triterpenoid saponin isolated from the roots of Clematis chinensis with antioxidant and anti-inflammatory cardioprotection effects against MI/R injury, yet the anti-apoptotic effect and underlying mechanisms of AR in MI/R injury remain unclear. We hypothesize that AR may improve mitochondrial function to inhibit MI/R-induced cardiomyocyte apoptosis. In this study, we replicated an in vitro H9c2 cardiomyocyte MI/R model by hypoxia/reoxygenation (H/R) treatment. The viability of H9c2 cardiomyocytes was determined by MTT assay; apoptosis was evaluated by flow cytometry and TUNEL experiments; mitochondrial permeability transition pore (mPTP) opening was analyzed by a calcein-cobalt quenching method; and mitochondrial membrane potential (ΔΨm) was detected by JC-1. Moreover, we used western blots to determine the mitochondrial cytochrome c translocation to cytosolic and the expression of caspase-3, Bcl-2, and Bax proteins. These results showed that the application of AR decreased the ratio of apoptosis and the extent of mPTP opening, but increased ΔΨm. AR also inhibited H/R-induced release of mitochondrial cytochrome c and decreased the expression of the caspase-3, Bax proteins. Conversely, it remarkably increased the expression of Bcl-2 protein. Taken together, these results revealed that AR protects H9c2 cardiomyocytes against H/R-induced apoptosis through mitochondrial-mediated apoptotic signaling pathway.

• Authors: Haiyan Ding, Rong Han, Xueshan Chen, Weirong Fang, Meng Liu, Xuemei Wang, Qin Wei, Nandani Darshika Kodithuwakku, Yunman Li
• Journal: Molecules (Basel, Switzerland) (Molecules) Vol. 21 Issue 6 (May 30 2016) ISSN: 1420-3049 [Electronic] Switzerland
• PMID: 27248986 (Publication Type: Journal Article)

Chemical References

• Biomarkers
• Mitochondrial Membrane Transport Proteins
• Mitochondrial Permeability Transition Pore
• Protective Agents
• Proto-Oncogene Proteins c-bcl-2
• Saponins
• bcl-2-Associated X Protein
• clematichinenoside
• Cytochromes c
• Caspase 3

Topics

• Animals
• Apoptosis (drug effects)
• Biomarkers
• Caspase 3 (metabolism)
• Cell Line, Tumor
• Cell Survival (drug effects)
• Cytochromes c (metabolism)
• Hypoxia (metabolism)
• Ion Channel Gating
• Membrane Potential, Mitochondrial (drug effects)
• Mitochondria (drug effects, metabolism)
• Mitochondrial Membrane Transport Proteins (metabolism)
• Mitochondrial Permeability Transition Pore
• Myocardial Reperfusion Injury (metabolism)
• Myocytes, Cardiac (drug effects, metabolism)
• Protective Agents (chemistry, pharmacology)
• Proto-Oncogene Proteins c-bcl-2 (metabolism)
• Saponins (chemistry, pharmacology)
• Signal Transduction (drug effects)
• bcl-2-Associated X Protein (metabolism)