Protective effects of linoleic acid against E. coli lipopolysaccharide-induced oxidative and nitrosative toxicity in buffalo luteal cell culture.

backgroundThe current study aimed to investigate the varying concentrations of E. coli lipopolysaccharide (LPS) on the mitochondrial function of buffalo luteal cells (LCs), and evaluate the protective effect of linoleic acid (LA) concurrent with toxic doses of LPS on LC performance during primary culture.

methodsOvaries containing a corpus luteum (n = 21) were transported to the laboratory within 1-2 h after slaughter. After isolating LCs and verifying their viability with trypan blue staining, cells were pre-incubated. The first experiment was performed by exposure of LCs to varying amounts of LPS (0.25, 0.5, 1, 2, 4, 8, 16, and 32 µg/mL) in the 24-well culture plate. Mitochondrial function was assessed using the MTT assay at 0, 24, and 48 h of incubation. In the second experiment, LA (as conjugated with bovine serum albumin) at different concentrations (0.125, 0.25, 0.50, and 1 mM) were added to LCs media concurrent with constant toxic dose of LPS (8 µg/mL). Mitochondrial function, total antioxidant capacity (TAC), amounts of lipid hydroperoxide (LHPO), malondialdehyde (MDA), and total nitrate-nitrite (TNN) were assessed at 0, 24 and 48 h. Experiments 1 and 2 consisted of four independent repetitions and a quadruplicate design for each experimental group.

resultsCo-administration of LA, particularly at concentrations of 0.25 and 0.50 mM, with LPS enhanced mitochondrial function and increased TAC levels at 24 and 48 h (P < 0.05). Total LHPO was not affected by LA administration. The TNN index was significantly reduced by LA treatment at doses of 0.25, 0.50, and 1 mM at 48 h (P < 0.001). Similarly, the MDA index was significantly reduced by all doses of LA at both 24 and 48 h (P = 0.038).

conclusionLA could ameliorate the harmful effects of E.coli's LPS on buffalo LCs via the reduction of nitrosative and peroxidative toxicity, and upregulating the antioxidant levels during primary culture. These findings suggest that LA-enriched feed warrants investigation for its effects on buffalo cow conception during breeding periods.