Effects Of The Combination Of Potassium Sorbate And Saccharin Sodium On The Growth And Development Of Mice Small Intestinal Epithelial Cells

Food additive safety valuation standards are based on the toxic effects of a single food additive,whereas in the industry,a product often contains several additives.A variety of food additives together in the human body may have a combined effect of synergy,antagonism,which poses new challenges to the toxicity assessment of traditional food toxicology and food additives.In this study,the Caco-2 cell model and the mini-gut intestinal organ model were used to evaluate the toxicity of some common food additives application in the food industry.The main research contents and results are as follow:(1)The Caco-2 cell model was used as the research object,and the effects of five kinds of food preservatives such as potassium sorbate,sodium sulfite anhydrous,sodium pyrosulfite,sodium benzoate and dehydroacetic acid sodium salt and five artificial sweeteners such as sodium saccharin,sucralose,acesulfame K,sodium cyclamate and aspartame were evaluated.The cell proliferation inhibition toxicity was found to exhibit a dose positive correlation.The order of toxicity was: sodium pyrosulfite > sodium sulfite > potassium sorbate > dehydroacetic acid sodium salt > sodium benzoate,sodium saccharin > acesulfame K > aspartame > cyclamate > sucralose.Potassium sorbate and saccharin sodium,which are more commonly used in the food industry,were selected for joint experiments.The toxicity of cell proliferation was a synergistic combination through MTT method and 2×2 factorial analysis method.Using high content analysis system and qPCR technology,it was found that the combination of potassium sorbate and sodium saccharin could significantly reduce the number of cells,increase the release of reactive oxygen species,the fluorescence intensity of necrotic cells in the potassium sorbate group,the sodium saccharin group and the combined group were 125.72±7.35%,130.11±2.82%,and 176.41±8.54%,respectively.The total fluorescence intensity of ROS was 161.51%±7.47% and 144.16%±2.83,respectively.%,196.60% ± 6.91%.Moreover,it increased cell membrane permeability and decrease mitochondrial membrane potential,and highly express the mRNA levels of Bax,Caspase-3,Caspase-12,P53 and Cytochrome,low expression of Bcl-2 mRNA levels.At the same time,the sodium sulfite and cyclamate with antagonistic combination were used for comparison verification.(2)The toxicity of these ten food additives were detection based on the mini-gut cell model.They were found to reduce the area growth rate,budding rate,number of sprouts and cell viability of mini-gut to varying degrees.Among all,potassium sorbate,sodium sulfite,sodium dehydroacetate,sucralose,acesulfame,cyclamate and aspartame had a significant dose-positive toxicity to the cell viability of mini-gut,with dose increase cell viability reduction.The morphology and some biological characteristics and cell viability of mini-gut were examined.It was found that the combination of potassium sorbate and sodium saccharin could significantly inhibit the growth of mini-gut.Potassium sorbate and sodium saccharin,sodium sulfite and cyclamate affected the differentiation of mini-gut cells,resulted in increasing stem cells,decreasing endocrine cells,and increasing the absorption of cells.The level of oxidative stress in Mini-gut also changed,in which the activity of superoxide dismutase and the activity of glutathione peroxidase decreased.At the same time,the mRNA expression of Bcl-2l11,caspase-3 and Fas was significantly increased by qPCR.The oxidative stress pathway may also be a signal pathway for food additives to affect mini-gut growth.Potassium sorbate and sodium saccharin have synergistic effects,and sodium sulfite and cyclamate may be antagonistic effects.This study showed that mini-gut based on intestinal organs is a good model for evaluating the toxicity of food additives,especially joint effects.Potassium sorbate and sodium saccharin alone and in combination can cause cytotoxicity to intestinal cells and induce apoptosis.The combination of potassium sorbate and sodium saccharin was a synergistic toxic effect,and the combination of sodium sulfite and cyclamate exhibited an antagonistic toxic effect.