Protocols for analyzing metabolic derangements caused by increased NADH/NAD+ ratio in cell lines and in mice

Shanshan Liu; Yan Ma; Hui Jiang

doi:10.1016/j.xpro.2021.101120

Protocols for analyzing metabolic derangements caused by increased NADH/NAD⁺ ratio in cell lines and in mice

STAR Protoc. 2022 Jan 19;3(1):101120. doi: 10.1016/j.xpro.2021.101120. eCollection 2022 Mar 18.

Authors

Shanshan Liu¹, Yan Ma¹, Hui Jiang¹

Affiliation

¹ National Institute of Biological Sciences, Beijing 102206, China.

Abstract

Mitochondrial electron transport chain (ETC) dysfunction elevates the NADH/NAD⁺ ratio to cause metabolic derangements. Here we describe a protocol to measure the NADH/NAD⁺ ratio and analyze the rewiring of glucose metabolism using [4-²H]-glucose, [3-²H]-glucose, and [U-¹³C]-glucose in ETC-inhibited human cancer cells. We also describe a protocol to analyze the NADH/NAD⁺ ratio-sensitive metabolites in mouse plasma and mouse liver following phenformin treatment. These protocols comprehensively analyze the metabolic derangements resulting from increased NADH/NAD⁺ ratio in in vitro and in vivo models. For complete details on the use and execution of this profile, please refer to Liu et al. (2021).

Keywords: Cell Biology; Cell culture; Cell-based Assays; Mass Spectrometry; Metabolism; Model Organisms.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Line
Glucose / metabolism
Metabolic Diseases*
Mice
NAD* / metabolism
Oxidation-Reduction

Substances

NAD
Glucose