| Objective:The percentages of people with type2diabetes have rapidly increased throughout our Country. Researchers proposed that high morbidity of the type2diabetes is associated with catch-up growth. It is known that diet induced GLP-1secretion is impaired in obesity and type2diabetes whose blood free fatty acids are higher than non-obese people. People and animals with catch-up growth show disproportion lipid accumulation、elevated free fatty acids and impairment of nutrition stimulated GLP-1secretion. Elevated free fatty acids can impair cell function and promote cell death by its toxic effects which play an important role in abnormal glucose metabolism. Palmitic acid is a common substance used to research lipotoxicity effects on cell in vitro. Here we reported the impact of palmitic acid on the viability and enzymes involved in lipid metabolism of human intestinal endocrine L cell line (NCI-H716).Methods:The NCI-H716cells was cultured in RPMI1640medium supplemented with10%FBS、100IU/ml penicillin and100μg/ml streptomycin. Before the experiments, cell medium was refreshed with total medium. The day before the experiment cells were seeded in the culture plates coated with matrigel. Twenty-hours later, the cells were divided into four groups:1cell viability of the NCI-H716cells under the conditions of different concentration of palmitic acid and different treatment time:NC group (normal control group)、PL group (with0.125mM palmitic acid)、PM group (with0.25mM palmitic acid)、PH group (with0.5mM palmitic acid). The cell viability was detected by cell counting kit-8(CCK-8)after treatment for four hours、twenty-four hours or forty-eight hours, separately.2After forty-eight hours treatment with different concentrations of palmitic acid, the mRNA expression of NCI-H716cells regarding lipid metabolism such as fatty acid synthase (FAS), acetyl-CoA carboxylase(ACC)、 diacylglycerol acyltransferase-1(DGAT-1)、 carnitine palmitoyltransferase-1(CPT-1) were detected by RT-PCR.Results:1Treatment of NCI-H716cells with palmitic acid resulted in a significant decrease in cell proliferation in a concentration dependent manner:compared with control group, cell viability was decreased by13%(P<0.05) or53%(P<0.01) in the PM or PH group after four hours treatment with palmitic acid. There was no statistically significant in the PL group when compared with NC group. Compare with the NC group, stimulation with palmitic acid in PL group for twenty-four or forty-eight hours, the cell viability was decreased by53%or68%; the PM group decreased by72%or76%and the PH group decreased by78%or79%. All of the P-value<0.01.2The RT-PCR showed:Treatment of NCI-H716cells with palmitic acid for forty-eight hours, the ACC mRNA was decreased by11%(P>0.05)、18%(P<0.05)、37%(P<0.01) in the PL、PM or PL group compared with NC group. As in the same conditions, mRNA of the CPT-1was increased by28%(P>0.05)、77%(P<0.01) or100%(P<0.01),separately. The expressions of the FAS mRNA have the trend to decrease along with the level of the palmitic acid, but there is no statistically significant between the four groups. The expression of the DGAT1among the NC group and the palmitic acid groups had no obviously change.Conclusion:Palmitic acid stimulation can impair the viability of the NCI-H716cells in a concentration dependent manner. The expression of the enzymes mRNA involved in the lipid metabolism were changed by the palmitic acid dispose, which may play a vital role in palmitic acid induced cell damage. |
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