Kangxian Yixin Prescription Interferes with Mitochondrial Permeability Transition Pore to Inhibit Cardiomyocyte Apoptosis / 中国实验方剂学杂志

ABSTRACT

Objective:To explore the mechanism of Kangxian Yixin prescription in regulating mitochondrial permeability transition pore（mPTP）and inhibiting cardiomyocyte apoptosis. Method:H9c2 cardiomyocytes were cultured routinely. After 8 h of starvation，the cells were divided into the normal group，model group，Kangxian Yixin prescription（0.25 g·L^-1） group，and cyclosporin A（CsA，10 μmol·L^-1） group and treated with the corresponding drugs for 24 h for follow-up experiments. The H9c2 cardiomyocyte hypertrophy model was induced by norepinephrine（NE），whose optimal concentration was determined by real-time polymerase chain reaction （Real-time PCR）. The degree of mPTP opening was detected by flow cytometry， followed by the measurement of mRNA and protein expression levels of apoptosis-related factors cyclophilin D（Cyp-D），cytochrome C（Cyt-C），and cysteine aspartate-specific protease-3（Caspase-3） after mPTP opening and the quantification of mitochondrial membrane potential. Result:When the concentration of NE was 200 μmol·L^-1， the mRNA expression levels of atrial natriuretic peptide（ANP） and brain natriuretic peptide（BNP） were the highest， implying that it was the optimal concentration to induce H9c2 cell hypertrophy. Compared with the normal group，the model group exhibited excessive opening of mPTP，weakened relative fluorescence intensity in mitochondria， decreased mitochondrial membrane potential（<italic>P</italic><0.05，<italic>P</italic><0.01），and elevated mRNA and protein expression of Cyp-D，Cyt-C，and Caspase-3（<italic>P</italic><0.05）. Compared with the model group，both Kangxian Yixin prescription and CsA inhibited mPTP opening，enhanced the relative fluorescence intensity of mitochondria， increased mitochondrial membrane potential（<italic>P</italic><0.05，<italic>P</italic><0.01），and lowered the mRNA and protein expression of Cyp-D，Cyt-C，and Caspase-3 （<italic>P</italic><0.05）. Conclusion:Kangxian Yixin prescription inhibits cardiomyocyte apoptosis possibly by regulating mPTP opening and inhibiting the expression of apoptosis-related factors Cyp-D，Cyt-C， and Caspase-3.