Citation:
YUAN Lian, LIU Yu-Jiao, HE Huan, JIANG Feng-Lei, LI Hui-Rong, LIU Yi. Microcalorimetric Analysis of Isolated Rat Liver Mitochondrial Metabolism under Different Conditions[J]. Acta Physico-Chimica Sinica,
;2018, 34(1): 73-80.
doi:
10.3866/PKU.WHXB201707043
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Isolated rat liver mitochondria were proposed as a model to monitor real-time heat metabolism.A high-throughput and sensitive thermal activity monitor III (TAM III) was used to detect the P-t curves of mitochondria under different conditions,including different mitochondrial concentrations,different substrates,different buffers,respiratory inhibitors,Ca2+,and CsA.We determined the thermokinetic parameters through calculation.The results showed that:(1) higher concentration of mitochondria led to faster energetic metabolism;(2) when succinate was the direct respiratory substrate,it promoted mitochondrial metabolism,in contrast to the condition when an indirect substrate,pyruvate,was used;(3) high concentration of Ca2+(2.5 mmol·L-1) stimulated mitochondrial metabolism,however CsA,an inhibitor of mitochondrial permeability transition pores,could not reverse the Ca2+-induced mitochondrial alteration;(4) mitochondria in various buffers displayed different rates of heat metabolism,because of the different composition of the buffers;(5) mitochondrial metabolism was inhibited by respiratory inhibitors,especially NaN3,which is an inhibitor of Complex IV and which completely stopped the mitochondrial heat release.
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