Clinical Trial And Mechanism Study Of Xingxiao Pills In The Treatment Of Advanced Non-small Cell Lung Cance

Background:Non-small cell lung cancer is currently the leading cancer killer with extremely high morbidity and mortality.For advanced NSCLC patients,most of them are treated with chemotherapy drugs,but they probably lead to adverse reactions such as nausea,vomiting,and pain.Xing xiao Pill has the functions of warming yang and dissipating stasis,promoting blood circulation and removing blood stasis,detoxifying and relieving pain.It is a classic anticancer formula of traditional Chinese medicine.The application of Xing xiao Pills in malignant tumors has been published in books,but it has not been supported by clinical evidence,and the specific molecular mechanism is still unclear.Therefore,this study designed clinical and basic research to evaluate the clinical efficacy of Xing xiao Pills in the treatment of advanced NSCLC after chemotherapy,and to provide powerful proof evidence for its clinical application.Purposes:1 This research designed the clinical efficacy observation of Xing xiao Pill in the treatment of stage ⅢB-Ⅳ non-small cell lung cancer after chemotherapy,through which can evaluate the safety of the treatment of patients with traditional Chinese medicine syndromes,clinical efficacy.2 Through network pharmacology research,this research explored the targets and signal pathways of Xing xiao Pills in the treatment of lung cancer,aiming to support further research.3 This research designed an experimental study including animal and cell experiment.In vivo experiment aimed to explore the intervention effect and potential mechanism of Xing xiao Pill on lung cancer transplanted mice.And In vitro experiments were conducted to explore the water extract of Xing xiao Pill based on PI3K/AKT/mTOR pathway regulated fatty acid metabolism of A549 cells,thereby affecting the biological processes such as proliferation,migration and apoptosis of A549 cells..Method:1 Clinical observational research:The research was designed as a randomized controlled study with a trial scheme,recruited subjects in the trial accordance with the established inclusion and exclusion criteria.The trial was randomly divided into treatment group and control group.The control group was given routine treatment after chemotherapy,and the treatment group was given Xing xiao Pill orally for 2 weeks on the basis of the control group.The total observation period was one month,with the every about fourteen days of follow-up period.In addition,the trial observed the baseline data,traditional Chinese medicine syndrome,quality of life scale,WHO anti-cancer toxicity scale and safety indicators.Finally,SPSS 20.0 was used for statistical analysis.2 Network pharmacology research:The research obtained the main chemical components and targets of Xing xiao Pills through BATMAN-TCM database,screened effective compounds according to Swiss ADME,and predicted their targets through Swiss Target Prediction.Through Gene Cards and others,the trial obtained the main targets of lung cancer and integrates them.The research obtained the intersection targets of Xing xiao Pills in the treatment of lung cancer through Venn,used the String platform to obtain the intersection targets,and BisoGenet3.0 was used to screen the core targets of Xing xiao Pills in the treatment of lung cancer.Moreover,the research used the Metasacpe database to analyze the intersection targets,and the biological processes and pathways involved in the dots were analyzed.Cytoscape 3.7.2 software was used to construct the network of "active ingredientstarget-pathway" of Xing xiao Pills.Finally,Autodock was used for molecular docking verification.3 Experimental study:(1)In vivo experiment:The Lewis lung cancer xenograft mouse model was constructed and divided into Xing xiao Pill low-dose,medium-dose,high-dose,and cisplatin groups.Each group was given fourteen days of intragastric administration,and measuring the mouse tumor tissue every three days,including their volume and visceral coefficient.Besides,HE was responsible for observing the pathological changes of tumor tissue in each group,WB were in charge of detecting the effect of Xing xiao Pill regulating PI3K/AKT/mTOR signaling pathway,transcriptome sequencing was performed on the tumor tissues of mice in the highdose intervention and control groups of Xing xiao Pills,which is to analyze the differential genes,main biological functions and signaling pathways of Xing xiao Pills on lung cancer mice,and integrated with the results of network pharmacology research.(2)In vitro experiments:CCK-8,cell apoptosis,cell cycle and transwell test were in charge of detecting the inhibition rate,the effects on apoptosis and cycle and the consequence on the migration of A549 cells.Oil red O staining,TC,TG,HDL-C,LDL-C and free fatty acid kits were responsible for checking the outcome on lipid droplets,measuring the assay of TC,TG,LDL-C,HDL-C and free fatty acid in A549 cells intervened by Xing xiao Pills.Otherwise,WB were responsible for evaluating the results of A549 cells on the protein contents of PI3K/AKT/mTOR/SREBP1 pathway and FASN contents in A549 cells.Result:1 Observational study of clinical efficacy:(1)Efficacy evaluation:Traditional Chinese medicine syndrome score showed that compared with conventional treatment alone,after taking Xing xiao Pill orally for two weeks,the total traditional Chinese medicine syndrome score had a markedly decrease,and symptoms such as aversion to cold,cold extremities and other symptoms were significantly reduced;After one month,the treatment group significantly improved the total score of traditional Chinese medicine syndromes,relieved symptoms such as fear of cold,chills,constipation,chest tightness and chest pain,dark purple tongue,and the curative effect was better than that of the control group.Besides,the effective rate of traditional Chinese medicine syndrome was significantly improved in the treatment group.Quality of Life Scale score found that after two weeks of treatment,the overall healthy living status of treatment group was better than that of those who only received conventional treatment,moreover,after one month,the overall healthy living status,symptoms and specific symptom scores were all obtained improve and enhance.WHO anticancer drug acute and subacute side effects showed that after two weeks,compared to the control group,the constipation and pain of the treatment group were improved,and the other targets did not change significantly.(2)Safety evaluation:There were no obvious variety diversity in stool routine,blood,coagulation,urine,biochemistry and electrocardiogram examinations between the two groups during the trial.2 Network pharmacology research:The main pathways of Xing xiao Pill in the treatment of lung cancer included PI3K/AKT/mTOR,MAPK,MicroRNA,etc.The core targets were AKT1,SIRT7,MDM2,mTOR,etc.3 Experimental study:(1)In vivo experiment:Xing xiao Pill high-dose and cisplatin group could inhibit tumor growth in mice with lung cancer,and could make tumor tissue necrotic.Moreover,it could down-regulate the protein content of p-AKT.Transcriptome sequencing showed that the main signaling pathways of Xing xiao Pill in the treatment of lung cancer mice involved estrogen signaling pathway,PI3K/AKT/mTOR,PPAR,AMPK,aerobic glycolysis,fatty acid metabolism and other pathways.Integrated with the results of network pharmacology,they jointly focused on the PI3K/AKT/mTOR pathway and lipid metabolism way.(2)In vitro experiments:The IC50 group of Xing xiao Pills water extract made a role in inhibiting the proliferation and migration of A549 cells,abducted their apoptosis,and arrested the cell cycle in S phase.It could cut down lipid droplets,TC,LDL-C,TG and free fatty acid in cells.The mechanism involved is that it can down-regulated the protein expression levels of p-PI3K,AKT,p-AKT,p-mTOR,SREBP1 and FASN.Analysis conclusion:1 Observational study of clinical efficacy:(1)Efficacy evaluation:Xing xiao Pill could effectively improve the sensitivity to cold,cold limbs,constipation,chest tightness and chest pain,and dark purple tongue in advanced NSCLC after chemotherapy;Besides,it significantly improved the overall life and health status,symptoms and specific symptoms of patients;Moreover,it could improve constipation and pain of patients.(2)Safety evaluation:Xing xiao Pill has good safety in the treatment of NSCLC patients after chemotherapy.2 Network pharmacology research:The study initially revealed that Xing xiao Pills in the treatment of lung cancer may involve tumor angiogenesis,metastasis,metabolic reprogramming and other mechanisms.The main targets are AKT,mTOR,SIRT7,and the pathways involved include PI3K/AKT/mTOR,MAPK,etc.,which provided some frame of mind for future clinical application and basic research.3 Experimental study:(1)In vivo experiment:Xing xiao Pill had the effect of inhibiting tumor growth in mice with lung cancer transplanted tumor,and can accelerate tumor tissue necrosis.And also the expression of PI3K/AKT/mTOR can be down-regulated.Transcriptome sequencing and network pharmacology research showed that the main signaling pathways of Xing xiao Pill in the treatment of lung cancer is PI3K/AKT/mTOR,and fatty acid metabolism is the main mechanism.(2)In vitro experiments:Xing xiao Pills could inhibit the proliferation and migration of A549 cells,could block their cell cycle in S phase and expedite their apoptosis.Besides,Xing xiao Pill could reduce the content of lipid droplets,TC,TG,LDL-C,free fatty acid.The possible mechanism involved was that Xing xiao Pill inhibited fatty acid metabolism of A549 cells by down-regulating the expression level of PI3K/AKT/mTOR/SREBP1 pathway and FASN,which can inhibit the proliferation and migration of A549 cells,induce its apoptosis,and arrest its cycle in S phase.