| Background and purposeAs one of the "four major national medicines",Chinese medicine rhubarb has a high clinical frequency in clinical practice.The diarrhea effect of rhubarb mainly depends on the ingredients of anthraquinones.In recent years,studies have shown that long-term use of anthraquinones can lead to colon damage and colonic melanosis,and further investigation is needed to determine whether the cancer risk is present.Intestinal microbes are one of the important components of the microenvironment in the intestine tract and the destruction of intestinal microbial homeostasis promotes the development of intestinal inflammation and hyperplasia in intestinal epithelium.There are many articles showing the use of rhubarb altered the composition of the microbes.Therefore,this study selected the representative anthraquinones emodin in rhubarb and studied the role of emodin in the development of colitis and colonic hyperplasia from the perspective of intestinal flora.Research contents and resultsIn this study,C57BL/6 mice were used for the experiment.The drug-administered group was given 100 mg/kg emodin for 12 weeks,the AOM/DSS model control group was modeled with AOM,and all animals were intraperitoneally injected with AOM(10 mg/kg).One week after AOM injection,2%DSS was given for 5 consecutive days for modeling and rest for 10 days.The same DSS administration mode was repeated three times in total.The DSS model control group was given 2%DSS for 5 days for modeling and rest 10 In the same day,the same DSS administration mode was repeated three times.Animal feces were taken weekly for 16SrDNA detection and fecal inflammation index LCN2.Animals were weighed once a week.After 12 weeks of animal administration,the animals were treated.The animals were sacrificed before sacrifice.The animals were sacrificed and the liver was weighed.The spleen was weighed and taken,colonized,HE stained,CD45 immunohistochemical staining,β-catenin immunohistochemical staining,Ki67 And TUNEL immunofluorescence staining,using Luminex liquid chip detection technology to detect inflammatory factors in the blood,taking mesenteric lymph nodes,flow cytometry test for Th1,Th2 and Th17.The results showed that the emodin/AOM/DSS group had more severe inflammation than the model group,and there was a significant difference in serum CXCL1 and CXCL2,but at the same time compared with the model,the intestinal abnormality of the emodin group.There was a significant decrease in proliferation and a decrease in proliferation-related indicators,and there was no significant difference in Th1,Th2,and Th17.The above results show that long-term use of emodin promotes inflammation of AOM/DSS,but at the same time inhibits abnormal proliferation.At the same time,the DSS model was compared with the emodin administration group and the blank group.LCN2 results,HE staining,CD45 staining and serum inflammatory factors,and flow cytometry were used to detect Th1,Th2 and Th17,and the results showed that emodin was given.Significant colonic inflammation can be caused by the drug,in which CXCL1,CXCL2,TNF-α and IFN-y are significantly increased,and Thl is significantly different.Since emodin can cause inflammation in the colon,what role does intestinal microbe play in this process?The results of 16SrDNA sequencing of fecal samples taken during the experiment showed that there was a significant difference in the composition and diversity of the intestinal microbial flora in the emodin-administered group and the blank group,and the sequencing results were further excavated.The abundance changes of the flora were flodchange>2 and p-value<0.5 as the screening conditions.The results of screening the AOM/DSS module and emodin+AOM/DSS were compared with the results of the blank group and the emodin group.The desulfovibrio strains were screened out.The relevant data showed that the ectopicity of Vibrio cholerae was highly correlated with inflammatory factors such as CXCL1 and CXCL2.At the same time,the analysis results of the blank group and the emodin administration group showed polymorpha and rumenococcus.The abundance of short-chain fatty acids related to the production of short-chain fatty acids such as bacillus was significantly reduced,and the results of short-chain fatty acid experiments also showed a significant decrease in the yield of short-chain fatty acids.The results show that emodin has no effect on epithelial inflammatory factor secretion and proliferation.Based on previous results,short-chain fatty acids are important nutrients for intestinal epithelial cells and their production is decisive for the integrity of the intestinal epithelial mucosa.The dextran was used for the leakage test.The results of the in vivo imaging experiment showed that the mice in the emo-administered group showed obvious intestinal leakage,and the intestinal epithelial tight junction-associated proteins Claudin-1 and E-cadherin were significantly reduced.Finally,the immunological hybridization experiment showed that the emodin administration group showed an obvious ectopic flora.The above results show that the mucous layer of the intestinal epithelium is damaged.Before reviewing the results,emodin promotes inflammation and significantly inhibits the abnormal proliferation of intestinal epithelium,considering the important role of phosphorylation of STAT3 in the development of abnormal proliferation of colonic epithelial cells,and as an upstream protein of STAT3.β-catenin showed significant down-regulation in previous studies.Western blot and immunohistochemistry were used to detect p-STAT3.The results showed that emo significantly inhibited STAT3 phosphorylation,and q-PCR was used to correlate transcriptional factor Bcl2 downstream of STAT3.The results of Bcl211,Cyclin D1 and c-Myc showed that the expression and content of Bcl2,Bcl211,Cyclin D1 and c-Myc were significantly decreased.Finally,C57BL/6 nude mice were used for subcutaneous transplantation experiments to observe the inhibitory effect of emodin on the proliferation of colon cancer cells HT29.The results showed that the volume of subcutaneous xenografts in the emodin group was significantly smaller than that in the blank control group.The western blot results showed STAT3 phosphorylation and β-catenin.There is a significant decrease in β-catenin,and by these results we can obtain that emodin has a significant inhibitory effect on the proliferation of colon cancer cells.Conclusion and significanceBased on the above results,we found that emodin can promote the intestinal inflammation of the AOM/DSS model,and this process inhibits the abnormal proliferation of intestinal epithelial cells.Moreover,emodin can also directly cause colonic inflammation in animals,which is mainly caused by the alteration of intestinal microbial homeostasis and destruction of intestinal mucosal barrier.The inhibition of emodin proliferation is mainly through the phosphorylation of STAT3.From the perspective of gut microbiota,this topic clarifies that long-term use of emodin is a significant result of promoting the development of colitis.It also explains the mechanism by which emodin inhibits abnormal proliferation,the exploration of rhubarb medicinal properties,and the clinical use of TCM.It is of great significance. |
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