Acta Phys. -Chim. Sin. ›› 2018, Vol. 34 ›› Issue (1): 73-80.doi: 10.3866/PKU.WHXB201707043

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Microcalorimetric Analysis of Isolated Rat Liver Mitochondrial Metabolism under Different Conditions

Lian YUAN1,Yu-Jiao LIU1,Huan HE1,Feng-Lei JIANG1,Hui-Rong LI2,Yi LIU1,*()   

  1. 1 College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, P. R. China
    2 College of Life Science and Chemistry, Wuhan Donghu University, Wuhan 430212, P. R. China
  • Received:2017-06-13 Published:2017-11-01
  • Contact: Yi LIU E-mail:yiliuchem@whu.edu.cn
  • Supported by:
    the National Natural Science Foundation of China(21673166);Natural Science Foundation of Hubei Province, China(2015CFC892)

Abstract:

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.

Key words: Mitochondria, Microcalorimetry, Metabolic rate, Respiratory inhibitor, Mitochondrial permeability transition

MSC2000: 

  • O642.3