| Food additives are widely used in various foods to maintain food taste,color and other qualities.However,if used improperly or over the standard,it will cause harm to the human body.Today,sulfite is often added to food to prevent the oxidation of nutrients and inhibit the growth of microorganisms,and has long been used as a bleaching agent in the food industry.The aim of this study was to study the toxic effects of sodium sulfite as a food additive,liver cells have detoxification effect in human body,and their damage is also harmful to human health,in this study,human normal hepatocytes?L02?were used to analyze the damage of sodium sulfite to hepatocytes from a cellular perspective,and the genes differentially expressed in L02hepatocytes after ingestion of sodium sulfite were analyzed by transcriptomics,from these differentially expressed genes,the toxic mechanisms of sodium sulfite damage to human hepatocytes are elucidated.The main findings are as follows:1.Effects of Na2SO3 on morphology and proliferation of L02 cell?1?Effects of Na2SO3 on the morphological changes of L02 cells:Treatment of L02 cells with Na2SO3 at concentrations of 0 M,10-55 M,10-44 M,10-33 M,and 10-22 M for 24 h,48 h,and 72 h,respectively.The cell morphology of the control group was observed under an inverted microscope,however,the cells treated with different concentrations of Na2SO3 showed different degrees of clustering,membrane thinning and even cell fragmentation,and this phenomenon became more and more obvious with the increase of the treatment days.The higher the concentration of Na2SO3,the shorter the survival time of the cells and the stronger the toxicity to the cells.?2?Effect of Na2SO3 on L02 cell activity:Treat L02 cells with Na2SO3 at concentrations of0 M,10-55 M,10-44 M,2.5×10-44 M,5×10-44 M,5×10-33 M,and 10-22 M,and the activity of L02 cells treated with different concentrations of Na2SO3 for 72 hours was detected,the half inhibitory concentration?IC50?of Na2SO3 was 4.68×10-44 M.?3?The effect of Na2SO3 on the proliferation of L02 cells:different concentrations of Na2SO3?0,10-55 M,10-44 M,5×10-44 M,5×10-33 M,and 10-22 M?were used to treat L02 cells for 0 h,24 h,48 h and 72 h respectively,and with the increase of time,the more obvious inhibition effect,and low concentration of Na2SO3 treatment on L02 cells had no obvious effect.2.Effect of Na2SO3 on apoptosis of L02 cellL02 cell were treated with different concentrations of Na2SO3?0,10-55 M,10-44 M,10-33 M,and10-22 M?for 24 h,48 h and 72 h respectively.The apoptosis of L02 cells was observed by AO/EB double staining.The results showed that the number of normal cells with green fluorescence decreased gradually,while the number of apoptotic cells with orange fluorescence and red fluorescence increased greatly.In addition,L02 cells were treated with Na2SO3 at concentrations of 0 M,10-55 M,10-44 M,5×10-44 M,and 10-22 M for 72 h,and L02 cells were treated with half lethal concentration of Na2SO3 for different time.The apoptosis of L02 cells was detected by flow cytometry,the results showed that with the increase of Na2SO3 concentration and the treatment days,the number of normal cells decreased,but the number of early apoptosis,late apoptosis and cell necrosis rate increased significantly.3.Effect of Na2SO3 on differentially expressed genes in L02 cellThe cells treated with semi-lethal Na2SO3 for 72 hours?IC50 in the treatment group?and the cells treated without Na2SO3 for 72 hours?MOCK in the control group?were analyzed by transcriptomics.There were 2152 genes specifically expressed in the treatment group and 5075genes specifically expressed in the control group.There were 97 differentially expressed genes in the volcano map,p-adjust<0.05.Go analysis showed that most of the differentially expressed genes were classified as cell processes,single tissue processes,and metabolic processes.The results of KEGG analysis showed that most of the differentially expressed genes were closely related to metabolic pathway,cancer,signal transduction,apoptosis and inflammation.The result of qRT-PCR was consistent with that of transcriptome analysis. |
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