A Study Of The Effectiveness And Its Mechanism Of Shenling Baizhu Power In Treating Pyrotinib-induced Diarrhea In HER2-positive Breast Cancer

Objective: Pyrotinib is a novel epidermal growth factor receptor/human epidermal growth factor receptor-2(HER2)tyrosine kinase inhibitor that has demonstrated clinical efficacy in patients with HER2-positive breast cancer and HER2 mutant/amplified lung cancer.However,common adverse reactions,especially diarrhea,limit the clinical use of pyrotinib and need to be addressed urgently.Shenling Baizhu Powder is a traditional Chinese medicine compound,which has good therapeutic effect on gastrointestinal toxicity,but it has not been reported in the treatment of pyrotinib-related diarrhea.Therefore,we studied the therapeutic effect of Shenling Baizhu Powder on diarrhea caused by pyrotinib,and studied its possible mechanism,providing a theoretical basis for the clinical use of Shenling Baizhu powder in the treatment of pyrotinib-induced diarrhea.Methods: First,we compared the effects of different doses of pyrotinib(20,40,60,and 80 mg/kg)on Wistar rats in order to establish a suitable and repeatable animal model,and we used this model to test the enterotoxicity of pyrotinib.Then,appropriate dose was selected to construct the diarrhea model of rats induced by pyrotinib,and Shenling Baizhu Powder was given daily(3.6 g/kg/bid for 10 days).The weight,diarrhea rate and diarrhea index of rats were recorded to evaluate the efficacy of Shenling Baizhu Powder.We then used 16 S r RNA sequencing to detect the intestinal microbiota of rats,collected metabolic profiles using non-targeted metabolomics methods,and identified key metabolic pathways using Metabo Analyst 5.0.The antitumor effect of Shenling Baizhu Powder on pyrotinibtreated cells was determined by CCK-8 method.Finally,network pharmacology and molecular docking techniques were used to predict and verify the target and pathway of action of Shenling Baizhu powder in the treatment of pyrotinib-induced diarrhea.Results: Rats developed recurrent diarrhea when given a daily dose of 80mg/kg pyrotinib.Using the animal model of pyrotinib-induced diarrhea,we verified that Shenling Baizhu Powder can significantly relieve pyrotinib-induced diarrhea,with a therapeutic effect of 66.7%.Shenling Baizhu Powder can regulate the disturbance of pyrotinib-induced microflora.LEf Se studies showed that SBP may decrease the relative abundance of Escherichia,Helicobacter and Enterobacteriaceae,and increase the relative abundance of Lachnospiraceae,Bacilli and Lactobacillales.In addition,25-hydroxycholesterol,guanidine acid,5-hydroxyindolyl pyruvate and c AMP were selected as potential biomarkers for Shenling Baizhu Powder in the treatment of pyrrotinib-induced diarrhea.In addition,Spearman analysis showed a correlation between intestinal flora and metabolites:the decrease of 25-hydroxycholesterol in pyrotinib + Shenling Baizhu Powder group was negatively correlated with Lachnospiraceae,but positively correlated with Escherichia and Helicobacter.Meanwhile,Shenling Baizhu Powder did not affect the inhibitory effect of pyrotinib on BT-474 cells and Calu-3 cells in vitro.Network analysis further revealed that Shenling Baizhu Powder can treat pyrotinib-induced diarrhea through multiple pathways,including inflammatory bowel disease,IL-17 signaling pathway,pathogenic Escherichia coli infection and c AMP signaling pathway.In addition,we identified 17 central genes based on network pharmacology,which is partially consistent with the findings of metabolomics.Molecular docking results showed that luteolin and quercetin,the main components of Shenling Baizhu Powder,have high affinity for key targets HIF1 AN,PDE3A and UGT1A1,and can regulate metabolic processes through these targets.Conclusion: We report for the first time a replicable pyrotinib-induced animal model,through which we verify that Shenling Baizhu Powder has an effective control against pyrotinib-induced diarrhea.In this study,we reported that Shenling Baizhu Powder could effectively treat pyrotinib-induced diarrhea by regulating intestinal flora and its related metabolites through microbiota sequencing and non-targeted metabolomics,and used network pharmacology to explore the key targets and mechanisms of Shenling Baizhu powder in the treatment of pyrotinib-induced diarrhea.This study provides data and theoretical support for the in-depth study of this mechanism and lays a foundation for clinical application.However,further systematic molecular biology experiments are still needed to further explore the molecular processes related to Shenling Baizhu Powder.