不同条件下离体大鼠肝脏线粒体能量代谢微量热分析

doi:10.3866/PKU.WHXB201707043

物理化学学报 >> 2018, Vol. 34 >> Issue (1): 73-80.doi: 10.3866/PKU.WHXB201707043

不同条件下离体大鼠肝脏线粒体能量代谢微量热分析

袁莲¹,刘玉娇¹,何欢¹,蒋风雷¹,李会荣²,刘义^1,*()

¹ 武汉大学化学与分子科学学院,武汉430072
² 武汉东湖学院生命科学与化学学院,武汉430212

收稿日期:2017-06-13 发布日期:2017-11-01
通讯作者: 刘义 E-mail:yiliuchem@whu.edu.cn
基金资助:
国家自然科学基金(21673166);湖北省自然科学基金项目(2015CFC892)

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 LIU^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)

摘要/Abstract

摘要：

为了探究线粒体的能量代谢过程，本文以离体大鼠肝脏线粒体为模型，利用多通道、高灵敏度的热活性检测仪TAM Ⅲ，实时监测了不同线粒体浓度、不同底物、不同缓冲液、几种呼吸抑制剂以及Ca²⁺和线粒体渗透转换孔抑制剂CsA存在时线粒体的能量代谢，获得了完整的热功率―时间曲线，并通过计算得到了线粒体能量代谢的热动力学参数。通过分析发现：（1）线粒体浓度越大，代谢越快；（2）直接底物琥珀酸钠使线粒体代谢更快；（3）高浓度Ca²⁺能够刺激线粒体快速产热，且在长期代谢进程中，线粒体渗透转换孔抑制剂CsA并不能改变Ca²⁺造成的影响；（4）不同缓冲液对线粒体代谢的影响基于其组分的不同，缓冲液中含有呼吸底物；（5）呼吸抑制剂都能抑制线粒体的能量代谢，尤其是复合物IV的抑制剂NaN₃，高浓度下使代谢停止。

关键词: 线粒体, 微量热, 代谢速率, 呼吸链抑制剂, 线粒体渗透转换

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

袁莲,刘玉娇,何欢,蒋风雷,李会荣,刘义. 不同条件下离体大鼠肝脏线粒体能量代谢微量热分析[J]. 物理化学学报, 2018, 34(1), 73-80. doi: 10.3866/PKU.WHXB201707043

Lian YUAN,Yu-Jiao LIU,Huan HE,Feng-Lei JIANG,Hui-Rong LI,Yi LIU. Microcalorimetric Analysis of Isolated Rat Liver Mitochondrial Metabolism under Different Conditions[J]. Acta Phys. -Chim. Sin. 2018, 34(1), 73-80. doi: 10.3866/PKU.WHXB201707043

链接本文: https://www.whxb.pku.edu.cn/CN/10.3866/PKU.WHXB201707043

https://www.whxb.pku.edu.cn/CN/Y2018/V34/I1/73

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不同条件下离体大鼠肝脏线粒体能量代谢微量热分析

Microcalorimetric Analysis of Isolated Rat Liver Mitochondrial Metabolism under Different Conditions

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