Studies On Differential Metabolites Of Glutamine And Glutamate On Mouse Fibroblasts And Human Cervical Adenocarcinoma Cells

The growth of cells requires a large supply of nutrients.As one of the nutrients,glutamine plays a crucial role in the growth and metabolism of cells.The use of glutamine in mouse fibroblast cells(L)and human cervical adenocarcinoma cells(HeLa)can be traced back to the 1950 s.Studies have shown that there are differences in the utilization of glutamine and glutamate between the two cells.This article examines the effects of glutamine and glutamate on L cells and HeLa cells,and establishes a method for cellular metabolomics analysis.This method was used to analyze the characteristics of metabolites and the changing rules of L cells and HeLa cells under culture conditions of glutamine or glutamate,and further explaining the reasons for the difference in the use of glutamine or glutamate.This article firstly validated the differences in the use of glutamine or glutamate in growth and proliferation of L cells and He La cells.The study found that the normal growth and proliferation of L cells and He La cells can only be maintained when the concentration of glutamine in the medium reaches 2 mmol/L or more.The replacement of glutamine with monosodium glutamate could not maintain the growth of L cells when the concentration is more than 10 times that of glutamine,but can maintain the growth of HeLa cells.Subsequently,the intracellular amino acid levels of both cells were determined and it was found that there were significant differences in the 11 kinds of amino acids contents between the two types of cells.Furthermore,at the level of enzyme activity,the effect of replacing glutamine with monosodium glutamate on the metabolism of glutamine and glutamate-related enzymes was investigated.The activity of glutamine metabolism-related enzymes in HeLa cells increased,and that of L cells had no significant change in corresponding enzyme activity.Second,this article established a cellular metabolomic analysis method.The linear range(0.02 ~ 200 mg/L),correlation coefficient(0.9949 ~ 0.9997),detection limit(0.001 ~ 0.002 mg/L)and the method recoveries(88.7% ~ 102.6%)of the five representative compounds were examined.The results showed that the method has high sensitivity and accuracy,and reliable information can be obtained when used in cellular metabolomics analysis.Lastly,the metabolomic characteristics and changes of L cells and HeLa cells under glutamine or glutamate culture conditions were studied by using cellular metabolomics analysis.Culture time was set to 0 h,12 h,24 h,48 h,and 72 h.The analysis results showed that there were significant differences in the intracellular metabolic profiles of L cells and HeLa cells under the three culture conditions of glutamine deficiency,glutamine addition,and monosodium glutamate.Through multivariate statistical methods,differential metabolites of L cells and HeLa cells under the above three culture conditions were screened out,mainly including sugars,bases,and organic acids.