不同条件下离体大鼠肝脏线粒体能量代谢微量热分析

引用本文: 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 shu

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 shu

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
基金项目:
国家自然科学基金（21673166）和湖北省自然科学基金项目（2015CFC892）资助

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:

English

Microcalorimetric Analysis of Isolated Rat Liver Mitochondrial Metabolism under Different Conditions

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 Date: 13 June 2017
Revised Date: 19 June 2017
Fund Project:
The project was supported by the National Natural Science Foundation of China (21673166) and 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:

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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

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Microcalorimetric Analysis of Isolated Rat Liver Mitochondrial Metabolism under Different Conditions

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通讯作者: 陈斌, bchen63@163.com

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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

English

Microcalorimetric Analysis of Isolated Rat Liver Mitochondrial Metabolism under Different Conditions

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

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CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

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CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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