Metabonomic Characteristics Of Fecal Supernatant In Patients With Ulcerative Colitis And Its Effect On Colonic Epithelial Cells

Objective:The effects of fecal supernatants in patients with ulcerative colitis on colonic epithelial cells were studied in vitro,and the metabolomic characteristics of fecal supernatants in patients with ulcerative colitis were analyzed.Methods:Forty patients with UC and forty healthy volunteers who met the criteria were enrolled into the UC group and the HC group,and fecal specimens were collected and FSN was prepared.Colonic epithelial cells NCM460 were treated with UC group and HC group FSN respectively.The effects of FSN on colonic epithelial cells NCM460 in vitro were evaluated by observing changes in cell morphology,measuring changes in cell viability,and detecting apoptosis.Ten cases of FSN in UC group and ten cases of FSN in HC group were selected respectively,and LC-MS detection was performed in both positive and negative modes to analyze the metabolomics characteristics of the FSN in UC patients.Results:FSN in patients with UC has a significant damaging effect on colonic epithelial cells NCM460.Intervention of NCM460 cells with FSN in UC group with a concentration of30% for 30 minutes caused significant cell contraction,separation and shedding.Compared with the HC group,the activity of NCM460 cells after FSN intervention in the UC group was significantly decreased(0.63 vs 0.89,P < 0.05).The damage of NMC460 cells in UC group was rapid and concentration-dependent.The higher the concentrationof FSN in UC group,the more significant the damage to NCM460 cells.However,the damage effect of FSN in UC group with a concentration of 30% on NCM460 cells was time-dependent.The longer the intervention time,the more serious the damage of NCM460 cells.FSN in patients with UC can induce apoptosis in NCM460 cells.Compared with mixed FSN in HC group,the percentage of apoptosis in NCM460 cells treated with mixed FSN in UC group after 30 min was significantly increased(56.37% vs2.37%,P < 0.05).From the visual comparison of the TIC chart,it can be found that there are some differences in the peak spectrum between the FSN in the UC group and FSN in the HC group.Multivariate statistical analysis was used to mine and identify differential metabolites between them.The results showed that there were more than 100 metabolites between the FSN in the UC group and the FSN in the HC group.Compared with the FSN in HC group,the FSN in the UC group has some harmful metabolites such as2-naphthylamine,glycocholic acid,and various forms of lysoPC(15:0,16:0,18:0,18:1[11Z] and 20:1 [11Z])and other content increased significantly.At the same time,?-linolenic acid,stearidonic acid,prostaglandin E2,riboflavin,pantothenic acid,nicotinic acid,coenzyme Q4,ubiquinone(Q2),adenosine and other metabolites with protective effects of the body are significantly reduced.Conclusion:FSN in patients with UC can cause damage to colonic epithelial cells,and serum may reduce its damage.At the same time,the metabolomics characteristics of FSN in patients with UC were significantly different from those in healthy people,and the content of some harmful metabolites increased significantly.