title: Polyunsaturated fatty acids incorporation into cardiolipin in H9c2 cardiac myoblast.
abstract: Docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), known as ω-3 polyunsaturated fatty acid (PUFA), are common nutrients in daily food intake and have been shown to prevent cardiovascular disease and improve cardiac functions. Cardiolipin is a mitochondrial phospholipid necessary for maintaining physiological function of mitochondria. Several studies have indicated that the cardiolipin acyl chain compositions affect the function of cardiolipin and mitochondria. Here, we investigated the structural changes of cardiolipin after DHA and EPA supplementation and compared them to arachidonic acid (AA) treatment. H9c2 cardiac myoblast was used as a cell model, and cardiolipin species was monitored and identified via LC-MS and MS/MS. Our results showed distinct mass envelopes of cardiolipin with the same carbon number but different double bonds in mass spectrum. There were 116 cardiolipin species with 36 distinct mass in 6 mass envelopes identified by MS/MS. Three days of PUFA treatment resulted in decreases of low-molecular-weight cardiolipin and increases of high-molecular-weight cardiolipin, suggesting the incorporation of exogenous DHA, EPA and AA into mitochondrial cardiolipin. PUFA incorporation was further verified by MS/MS analysis. More importantly, we found that DHA supplementation elevated the percent content of less unsaturated cardiolipin species and highly unsaturated cardiolipin species, containing ω-3 fatty acyl chains, indicating a ω-3 fatty acid incorporation mechanism with peroxidation protection. Our results indicate that PUFA supplementation differentially perturbed the fatty acyl chain compositions in the mitochondrial cardiolipin in the H9c2 cardiac myoblast, suggesting that mitochondrial membrane and the function of mitochondria are susceptible to exogenous lipid species.
authors: Department of Chemistry, Tunghai University, Taichung 40704, Taiwan.
authors: Hsiu-Chi
authors: HC
authors: Ting
authors: Department of Chemistry, Tunghai University, Taichung 40704, Taiwan.
authors: Yu-Jen
authors: YJ
authors: Chao
authors: Department of Chemistry, Tunghai University, Taichung 40704, Taiwan; Life Science Research Center, Tunghai University, Taichung 40704, Taiwan. Electronic address: howardhsu@thu.edu.tw.
authors: Yuan-Hao Howard
authors: YH
authors: Hsu
mesh_terms: D000818:Animals; D002308:Cardiolipins / Q000737:chemistry / Q000378:metabolism*; D002460:Cell Line; D002470:Cell Survival; D002851:Chromatography, High Pressure Liquid; D004042:Dietary Fats, Unsaturated / Q000009:adverse effects / Q000378:metabolism*; D019587:Dietary Supplements; D004281:Docosahexaenoic Acids / Q000009:adverse effects / Q000737:chemistry / Q000378:metabolism; D015118:Eicosapentaenoic Acid / Q000009:adverse effects / Q000737:chemistry / Q000378:metabolism; D005230:Fatty Acids, Nonesterified / Q000009:adverse effects / Q000737:chemistry / Q000378:metabolism; D015525:Fatty Acids, Omega-3 / Q000009:adverse effects / Q000737:chemistry / Q000378:metabolism*; D043371:Fatty Acids, Omega-6 / Q000009:adverse effects / Q000737:chemistry / Q000378:metabolism*; D008929:Mitochondria, Heart / Q000378:metabolism*; D008958:Models, Molecular; D015394:Molecular Structure; D008970:Molecular Weight; D032446:Myoblasts / Q000378:metabolism*; D009994:Osmolar Concentration; D051381:Rats; D053719:Tandem Mass Spectrometry
chemical_list: D002308:Cardiolipins; D004042:Dietary Fats, Unsaturated; D005230:Fatty Acids, Nonesterified; D015525:Fatty Acids, Omega-3; D043371:Fatty Acids, Omega-6; D004281:Docosahexaenoic Acids; D015118:Eicosapentaenoic Acid
keywords: Cardiolipin; DHA and EPA; Lipidomics; Mass spectrometry; Mitochondria; Polyunsaturated fatty acid