HBD Inhibits The Development Of Colitis-Associated Cancer In Mice Via The IL-6R/STAT3 Signaling Pathway

ObjecitveColitis-associated cancer(CAC)is a malignant disease of the colon that is caused by recurrent episodes of chronic intestinal inflammation.Huangqi Baizhu Decoction(HBD)is a classic prescription comprised of Radix Astragali and Rhizoma Atractylodis,which are usually used to treat digestive conditions,such as peptic ulcers,colitis or colorectal carcinoma in clinics.Therefore,we want to explore whether Huangqi Baizhu Decoction can slow down or reverse the occurrence and development of CAC by down-regulating IL-6R/STAT3 pathway.MethodsAOM was injected into abdominal cavity once,combined with 3%DSS free drinking for three cycles to make model in C57BL/6 mice using and treated with Huangqi Baizhu Decoction.After the end of modeling,the expression of IL-6R/STAT3 pathway-related proteins GP130,STAT3,p-STAT3,IL-6R,Cyclin D1 and Survivin protein were detected by Western-blot,and the expression position and expression level of p-STAT3,ki-67 and Jak2 were analyzed by immunohistochemistry.The mRNA changes of inflammatory factors TNF-a,IL-1? and IL-6 were analyzed by fluorescence quantitative PCR.MTT assay was used to detect the inhibitory effect of astragaloside IV and atractylenolide II on the proliferation of human colon cancer epithelial cells HCT-116 cells,and stimulated HCT-116 and mouse-derived macrophages RAW264.7 cells with IL-6.Astragaloside+Atractylenolide II was used to analyze changes in IL-6R/STAT3 pathway-related proteins using Western-blot.ResultsCAC mice developed symptoms such as lack of energy,reduced food intake,body weight loss,weight loss,and bloody diarrhea.At the end of modeling,the body weight of the model group was significantly reduced compared with the normal group;compared with the model group,the HBL,HBH and SASP groups significantly improved the survival rate of CAC mice;the mass/length of the colon of the model group mice The value was significantly higher than the normal group(P<0.01);the quality/length value of the HBL group decreased(P<0.01);the quality/length value of the SASP group decreased(P<0.05);the data showed that the average size of colon tumors in the model group and The number increased significantly,while the values in the HBL and SASP groups were significantly lower(P<0.05).Morphological observation of colonic tissue in CAC model(8w)showed that the mucosal glands were disordered,the goblet cells were significantly reduced,the inflammatory cells infiltrated in the mucosa and submucosa,and the moderately severe dysplasia in the distal mucosa of the colon.The drug group can significantly reduce the severity of glandular hyperplasia and the degree of inflammatory cell infiltration in the mucosa,which can significantly alleviate the symptoms caused by AOM/DSS.Immunohistochemistry results show that HBD can significantly inhibit proliferation of tumor cells,and decreased expression of Jak2 and p-STAT3-related proteins in IL-6R/STAT3 pathways;Western-blot results showed that HBD exerts anti-inflammatory effects by inhibiting IL-6R/STAT3 pathway,and by reducing expression of Cyclin D1 and Survivin inhibited the proliferation of tumor cells;The results of real-time PCR showed that the mRNA expression of TNF-a and IL-1? was significantly decreased after administration.In vitro,The MTT assay showed that Astragaloside+Atractylenolide II can significant inhibited cell viability in time-and dose-dependent manners.In addition,Astragaloside IV and Atractylenolide II notably suppressed the protein expression of IL-6R,STAT3,Cyclin D1 and Survivin induced by IL-6 in HCT-116 and RAW 264.7.ConclusionHuangqi Baizhu Decoction can exhibit anti-inflammatory and anti-proliferative effects by down-regulating the expression of IL-6R/STAT3 pathway protein,which can inhibit or slow the occurrence and development of CAC,and can be used as a drug for the treatment and prevention of CAC.