| ?Objective?In this study,colonic stem cell nest cell polarity was lost during the transformation of colitis cancer,which induced colonic crypt aberration to the point of entry.Choosing the AOM/DSS-induced mouse colitis cancer transformation model as the research object,on the basis of clarifying the compatibility of Wu-zhu-yu and Huang-lian to prevent the delay of mice colitis cancer transformation and the effect on colon distortion crypt foci,observe the different compatibility ratios.For mice,intestinal tissues,middle inflammation phases,related factors,sub-IL-6 and TNF-? expressions,and effects on Wnt-1/?-catenin signaling pathways are controlled.Node.Intestinal stem cell nest cell polarity,reducing the number of aberrant crypt foci from the perspective of the role of Wu-zhu-yu and Huang-lian to prevent the delay of colitis cancer transformation mechanism.?Methods?1.Establishment of AOM/DSS colitis cancer transformed mouse model: C57BL/6 mice were intraperitoneally injected with AOM at a dose of 10 mg / kg on the first day,and after 7 days were given 2.5% DSS solution for 7 consecutive days followed by sterile drinking water Continuous feeding for 14 days is a cycle,repeating 3 cycles.The colon tissues of mice were taken at the end of cycles 1,2,and 3,respectively,while monitoring the general status,diet,weight,stool,etc.of the mice every day.Observing the general shape of colon tissues after taking the materials,and obs.ervin.g the pat.holo.gy and dist.ort.ion of col.on tis.sues by H&E stain The n.um.ber of cry.pt foci.The establishment of the model was determined by the pathological staining of mice at 1,2,and 3 cycles and the number of crypt foci.2.The effect of Wu-zhu-yu and Huang-lian compatibility on AOM/DSS induced colitis cancer transformation model and ACF in mic.e: the m.ice w.ere ra.ndomly divi.ded in.to bla.nk gr.oup,mod.el gro.up,Yu-Li 1:1 group,Yu-Li 1:6 group,Yu-Li Even 6:1 groups,24 in e.ach g.roup.Exce.pt for th.e bla.nk gro.up,the mice in the other groups were given AOM/DSS to induce modeling.Each group began to give the corresponding drug intervention 7 days after the injection of AOM,and the drug was continuously administered for 3 cycles.Materials were collected at the end of each cycle to monitor the general status,weight change,DAI score,tumor formation,etc.of the mice.By observing the changes of colon histopathological morphology and ACF in each cycle,the efficacy of Wu-zhu-yu and Huang-lian combination was evaluated and screened Out of the best compatibility ratio of cornelian compatibility.3.The mechanism of Wu-zhu-yu and Huang-lian Compatibility preventing the delay of AOM/DSS-induced colitis cancer transformation in mice: part of the colon tissue of the mouse was cut,and the expression of inflammation-related factors IL-6 and TNF-? in the tissue was detected by ELISA.Real-time quantitative fluorescence PCR was used to det.ect the expre.ssi.on of W.nt-1 and ?-cat.en.in m RNA in c..olon ti.ssue,and Western-blot was used to detect the expression of Wnt-1 and ?-catenin protein in colon tissue.?Results?1.AOM / DSS was successfully used to induce the establishment of a stable mouse model of colitis cancer transformation: after drinking DSS,the mice began to lose weight,become mentally depressed,lose their hair,slow acti.vity,pe.rian.al blo.od stains,a.nd inc.rea.se t.he DAI sco.re.H&E pathological tissue staining showed that in the model group,at the end of the first cycle,tissue specimens: colon monolayer columnar epit.het.liu.m and la.mina pro.pria,exten.sive infl.ammato.ry cell inf.iltr.ation and inflamee.mat.ion form.ation were seen.Tissue specimens at the end of the second cycle: deformed glandular shape,disordered arrangement,division,partial distortion of crypt foci,and polymorphic moderate dysplasia in the colon of some mice.Tissue specimens at the end of the third cycle: atypical hyperplasia is very obvious,some areas of severe hyperplasia extend to the muscularis mucosa,most of the colon can see the distortion of the gland structure,the cell nuclei are deep stained,the proportion of nucleus and cytoplasm is significantly increased,and the colon tumor form.2.The 1:1 combination of Wu-zhu-yu and Huang-lian can indeed prevent the delay of AOM / DSS-induced colitis cancer transformation model in mice,and can reduce the formation of ACF: the general condition,mental state,activity,etc.of the Yu-Lian 1:1 group of mice The model is good,the percentage of weight gain is signi.fica.ntly increa.sed comp.ared wit.h the mo.del gro.up(p<0.01),and the tumor formation rate is lower than that of the other groups.Except for the blank group,the DAI scores of the other groups increased significantly during the period of DSS feeding,and the increase rate of DAI scores of the 1: 1 group of Yu-Lian was smaller than that of the other groups,There is no significant difference between the 6:1,1:6 group of Yu-lian and the model group.H&E pathological tissue staining showed that the destruction and hyperplasia of colonic crypts and colonies in the 1: 1 group of Yu-Lian at the end of the first,second,and third cycles were lighter than those of the other models,The pathological changes of Yu-lian 1:6,6:1 group were slightly better.The effect of Cornelian 1: 1 group on the ACF formation at the end of the second cycle was significantly reduced compared with the model group,the difference was significant(p<0.01),There is no significant difference in the groups of Yu-lian 6:1,1:6.3.Effects of Yu-lian 1:1 compatibility on IL-6,TNF-?,Wnt-1/?-catenin in colon tissue of mice with AOM/DSS-induced colitis cancer transformation: IL-6,TNF-? in mouse colon Content: Compared with the blank group,the model group mice.Serum IL-6,TNF-? rose.High,the difference was statistically significant(p<0.05);compared with the model group,Yu-lian 1:1 group,The content of IL-6 and TNF-? decreased,and the difference was statistically significant(p<0.05).It showed that the 1:1 compatibility of Yu-lian can down-regulate IL-6 and TNF-? in mouse serum.The expression of Wnt-1 and ?-catenin m RNA in the colon tissue of mice: Compared with the blank group of mice,the expression of Wnt-1 and ?-catenin m RNA in the colon tissue of the model group and Yu-lian 1:1 group were significantly increased(p<0.05).Compared with the model group mice,the expression of Wnt-1 m RNA and protein in colon tissue of Yu-lian 1:1 group was decreased(p<0.05);the difference between ?-catenin m RNA and protein was not statistically significant(p>0.05).?Conclusions?Compared with the 1:6 and 6:1 groups,the yu-lian 1:1 group has a better retarding effect in reducing the formation of ACF and preventing the delay of the AOM/DSSinduced transformation of mouse colitis to colon cancer.Its reduction in the number of ACF is related to the reduction of inflammation-related factors in the colon and intestine tissues of model mice,IL-6,TNF-?,and inhibition of Wnt-1/?-catenin signaling pathway. |
PDF Full Text Request