title: 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.affiliation: State Key Laboratory of Natural Medicines, Department of Physiology, China Pharmaceutical University, Nanjing 210009, China. dinghaiyan79@163.com.|College of Pharmacy, Xinjiang Medical University, Urumqi 830011, China. dinghaiyan79@163.com.
authors.forename: Haiyan
authors.initials: H
authors.lastname: Ding
authors.affiliation: College of Pharmacy, Xinjiang Medical University, Urumqi 830011, China. ronger051125@163.com.
authors.forename: Rong
authors.initials: R
authors.lastname: Han
authors.affiliation: College of Pharmacy, Xinjiang Medical University, Urumqi 830011, China. chenxueshan2010@sina.com.
authors.forename: Xueshan
authors.initials: X
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Natural Medicines, Department of Physiology, China Pharmaceutical University, Nanjing 210009, China. weirongfang@163.com.
authors.forename: Weirong
authors.initials: W
authors.lastname: Fang
authors.affiliation: Cancer Hospital Affiliated to Xinjiang Medical University, Urumqi 830011, China. liuxiaomeng198154@163.com.
authors.forename: Meng
authors.initials: M
authors.lastname: Liu
authors.affiliation: Xinjiang Key Laboratory of Medical Animal Model Research, Clinical Medical Research Institute of the First Affiliated Hospital of Xinjiang Medical University, Urumqi 830011, China. wxuemei1983@sina.com.
authors.forename: Xuemei
authors.initials: X
authors.lastname: Wang
authors.affiliation: Xinjiang Key Laboratory of Medical Animal Model Research, Clinical Medical Research Institute of the First Affiliated Hospital of Xinjiang Medical University, Urumqi 830011, China. fashion1027@163.com.
authors.forename: Qin
authors.initials: Q
authors.lastname: Wei
authors.affiliation: Institute of Indigenous Medicine, University of Colombo, Rajagiriya 11600, Sri Lanka. darshi_ko@yahoo.com.
authors.forename: Nandani Darshika
authors.initials: ND
authors.lastname: Kodithuwakku
authors.affiliation: State Key Laboratory of Natural Medicines, Department of Physiology, China Pharmaceutical University, Nanjing 210009, China. yunmanlicpu@hotmail.com.
authors.forename: Yunman
authors.initials: Y
authors.lastname: Li
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D015415:Biomarkers; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D045304:Cytochromes c / Q000378:metabolism; D000860:Hypoxia / Q000378:metabolism; D015640:Ion Channel Gating; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism*; D033681:Mitochondrial Membrane Transport Proteins / Q000378:metabolism; D000083162:Mitochondrial Permeability Transition Pore; D015428:Myocardial Reperfusion Injury / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000378:metabolism*; D020011:Protective Agents / Q000737:chemistry / Q000494:pharmacology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D012503:Saponins / Q000737:chemistry / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects*; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: D015415:Biomarkers; D033681:Mitochondrial Membrane Transport Proteins; D000083162:Mitochondrial Permeability Transition Pore; D020011:Protective Agents; D019253:Proto-Oncogene Proteins c-bcl-2; D012503:Saponins; D051028:bcl-2-Associated X Protein; C581539:clematichinenoside; D045304:Cytochromes c; D053148:Caspase 3
keywords: H9c2 cardiomyocytes; apoptosis; clematichinenoside (AR); hypoxia/reoxygenation; mitochondria; signaling pathway