Lipid metabolite analysis
Lipid metabolite analysis
Summary
Lipid metabolism is an important source of energy supply for the body, and an imbalance between lipid synthesis and catabolism triggers lipid metabolism disorders. Increased fatty acid synthesis and decreased oxidative catabolism lead to intracellular lipid accumulation. The accumulation of fat cells is the basis of obesity, but also an important risk factor for the occurrence of many diseases such as diabetes, atherosclerosis, tumors and cardiovascular disease. Currently, there are 2 main methods used to analyze lipid metabolites: fatty acid synthase activity assay and lipid content assay.
Principle
The basic principle of the lipid content assay is: fatty acidsynthase (FAS) is a key enzyme in fatty acid synthesis, which is abundant in mammalian liver, kidney, brain and mammary gland, as well as adipose tissues.The expression of FAS is at a low level in normal tissues, while the expression is abnormally high in malignant tumor cells. Acetyl CoA and malonyl CoA consume NADPH under the catalytic action of fatty acid synthase to produce long-chain fatty acids, so the decrease of NADPH is directly proportional to the activity and content of FAS, and the activity of FAS can be reflected by the decrease of absorbance of NADPH detected at 340nm.
Fat-soluble dyes are soluble in lipids in tissues and cells and are more soluble in lipids than in solvents. When a tissue section is placed in a dye solution, the dye leaves the solution and dissolves in the lipids (e.g., lipid droplets) within the tissue, giving the lipid droplets within the tissue an orange-red color. Oil red O belongs to the azo dye, is a very strong lipid solvent and lipid dye, and combined with triglycerides in the form of small lipid droplets, so it can be used for the analysis of the lipid content of tissues and cells.
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