Observation Of Iron Death Of Macrophages Mediated By ROS In UVAC Mice And Study On The Intervention Effect Of Sijunzi Decoction

Objective: By establishing a mouse model of ulcerative colitis associated carcinogenesis(UCAC),the phenomenon of ferroptosis in colon tissue macrophages and the changes of reactive oxygen species and ferroptosis-related protein PRXD6 were observed before and after Sijunzi decoction intervention.The influence of Sijunzi Decoction on the above changes provides experimental basis for the method of strengthening the spleen and nourishing Qi and the clinical application of Sijunzi Decoction.Methods: The 55 mice were divided into blank group,model group,western medicine group,low-dose Chinese medicine group,middle-dose Chinese medicine group,and high-dose Chinese medicine group by random number method.The UCAC mouse model was established by the1,2-dimethylhydrazine(1,2-dimethylhydrazine,DMH)/dextran sulfate sodium(DSS)method.The blank group and the model group were given equal doses of distilled water by gavage,and western medicine The group was given sulfasalazine by gavage,while the low,medium and high dose groups of traditional Chinese medicine were given corresponding doses(3.13g/kg,6.24g/kg,12.48g/kg)of Sijunzi Decoction by gavage for 4 consecutive weeks.After the experiment,fresh colon tissue was taken for HE staining to observe histopathology,transmission electron microscope to observe the ultrastructure of intestinal macrophages,ELISA to detect mitochondrial ROS content,Western-blot method to detect PRDX6 protein expression,RT-PCR method to detect PRDX6 m RNA expression.Results: 1.The UCAC mouse model was successfully established.The mice developed symptoms of diarrhea,mucus,pus and blood,and prolapse of the anus.Compared with the model group,the disease activity index(DAI)scores of each group decreased after treatment,and the DAI in the high-dose Chinese medicine group and the western medicine group The score decreased significantly(P<0.05).2.The pathology of the model group showed that the nuclei of the glandular cells in the mucosa were enlarged and hyperchromatic,and cell atypia was seen.After treatment,the pathological damage of colon tissue in the western medicine group and the traditional Chinese medicine groups were improved.In the tissue damage score,compared with the blank group,the tissue damage scores of each group were increased;compared with the model group,the tissue damage scores of each group were decreased;compared with the blank group,the tissue damage of the model group and the low-dose Chinese medicine group The academic score was significantly increased(P<0.05).3.Under the transmission electron microscope,ferroptosis in colon tissue macrophages can be observed in the model group,and the phenomenon of ferroptosis in colon tissue macrophages is reduced after adding western medicine and different doses of traditional Chinese medicine.4.ROS content: Compared with the blank group,the ROS content in the model group increased(P<0.05);compared with the model group,after adding western medicine and different doses of traditional Chinese medicine,the ROS content showed a downward trend,and the western medicine group had the most significant decrease(P<0.05).5.Compared with the blank group,the PRDX6 protein expression and gene expression in the model group were significantly up-regulated(P<0.05).Compared with the model group,after adding western medicine and different doses of traditional Chinese medicine,the PRDX6 protein expression and gene expression in the western medicine group and each dose group of traditional Chinese medicine showed a downward trend(P<0.05).Conclusion(s): 1.Sijunzi decoction can improve the symptoms of loose stool,mucus,pus and blood in UCAC mice to a certain extent;2.The content of ROS and the expression of PRXD6 in the colon tissue of UCAC mice increased,and Sijunzi decoction could reduce the content of ROS and the expression of PRXD6.3.ferroptosis can be observed in macrophages in colon tissue of UCAC mice,and Sijunzi decoction can inhibit the ferroptosis of macrophages.