Study Of Huangqin Decoction On Alleviating Intestinal Toxicity Of Irinotecan By Protecting Intestinal Mucosal Barrier

Background:Irinotecan(CPT-11)is an important chemotherapeutic drug in tumor treatment with strong inhibition effect on Topoisomerase I,an important enzyme involved in DNA replication.However,several side effects including delayed diarrhea,vomiting and nausea always occur with its clinical treatment,which limit its wide-spread applications.It has been reported that CPT-11 could cause injuries to the intestinal mucosa barriers with focus on epithelial cells damages,mucosa,proteins decreases,and microflora disturbance,which further lead to the aggravation of the intestinal toxicity.Huangqin Decoction(HQD)is a Chinese ancient prescription with a history of nearly two thousand years,and it is widely used in diarrhea and dysentery treatments.It was firstly recorded in "Shanghan Lun" written by Zhongjing Zhang,and the prescription consists of Astragalus,Licorice,Peony and Jujube.It has been published that HQD could restore the intestinal epithelium by promoting the regeneration of intestinal progenitor or stem cells,and it exhibited anti-inflammatory effects in mice by decreasing the infiltration of neutrophils or macrophages.According to the literature and our previous experiments,we proposed the following questions whether HQD can alleviate the delayed diarrhea through the protection of intestinal mucosal barrier injury caused by CPT-11.Thus,this study focused on the protection effects of HQD on intestinal mucosal barriers in the delay diarrhea induced by CPT-11.Restore effects of mucosal barriers consisting of intestinal epithelium,mucosal protein and intestinal flora were observed with the help of bioinformatics and immunological techniques in mice delay diarrhea model,the mechanism of reducing intestinal toxicity induced by CPT-11 by Huangqin Decoction was emphatically disclosed from the perspective of protecting intestinal mucosal barriers.The results will be helpful for clinical application of HQD in adjuvant therapy for tumor treatment with irinotecan.Objective: 1.To observe the recovery effects of HQD on intestinal epithelium damage in mice delayed diarrhea induced by CPT-11.2.To observe the adjust effects of HQD on mucosal protein decrease in mice delayed diarrhea induced by CPT-11.3.To observe the modulation effects of HQD on intestinal microflora disorder in mice delayed diarrhea induced by CPT-11.4.To reveal the mechanism of reducing intestinal toxicity induced by CPT-11 by HQD from the perspective of protecting intestinal mucosal barriers Methods: 1.The HQD extract was obtained by decocting method following freeze drying,and then resolved into different concentrations.2.The BALB/C mice were employed for mice delayed diarrhea model construction,mice were randomly divided into four groups: normal group(NC),irinotecan group(CPT-11),Huangqin decoction and irinotecan group(HQD+CPT-11)and Huangqin decoction Group(HQD).Mice of CPT-11 and HQD+CPT-11 groups were intraperitoneally injected with CPT-11(150mg/kg/day)from day 4 to day 7,while HQD+CPT-11 and HQD groups were treated with HDQ through gavage(3g/kg/day)from day 0 to day 9.The body weight was measured and the diarrhea symptom was observed daily.On day 9,mice were sacrificed,and the blood,small intestine,and feces were collected for the following experiments.3.Intestinal damage was detected by H&E staining,serum cytokine levels were detected by ELISA,TUNEL assay was used to detect apoptosis of intestinal mucosal cells,immunohistochemical staining and immunofluorescence were applied to detect intestinal mucosal barrier related proteins.4.The fecal flora was analyzed by PCR-DGGE assay followed 16 S rRNA sequencing,and the changes of flora between the groups were compared.Results: 1.HQD could effectively alleviate delayed diarrhea and other side effects(melena and weight loss)caused by CPT-11.2.HQD could reduce the intestinal damage by maintaining expression of the MUC2 and tight junction protein.3.HQD could modulate the intestinal microflora disorder induced by CPT-11 through adjusting the ratio of relative abundance of Proteobacteria and Lactobacillus.Conclusions:It could be concluded that the Huangqin Decoction can alleviate the intestinal toxicity by protection effects on intestinal mucosal barriers,which included restore of intestinal epithelium damage,maintaining expression of the MUC2 and tight junction protein,as well as adjustment of the intestinal microflora.The results will be helpful for clinical application of HQD in adjuvant therapy for tumor treatment with irinotecan.