The Research Of Material Basis And Mechanism Of Aconiti Lateralis Radix Praeparata Against NSCLC Based On Metabolic Reprogramming

Primary bronchial lung cancer(hereinafter referred to as lung cancer)has become the malignant tumor with the highest morbidity and mortality in my country,seriously endangering the health and lives of the general public.Non-small cell lung cancer(NSCLC)accounts for about 85%of all lung cancer patients,and the 5-year survival rate is only about 18%.Since surgery is not suitable for advanced patients,chemotherapy and radiotherapy have serious side effects,and targeted drug therapy is prone to drug resistance,conventional therapies have certain limitations in the clinical application of NSCLC.Traditional Chinese medicine classifies lung cancer into categories such as pulmonary accumulation and exhalation,and believes that it is based on yang deficiency,and when warming yang is the fundamental treatment,it is supplemented by regulating qi,eliminating phlegm,and removing blood stasis.Aconite is the first medicine for warming yang.It has the effects of regaining adversity,replenishing fire and assisting yang.The "Shen Nong’s Materia Medica" says that it can "break the symptoms and build up blood addiction." There have been studies and analyses in the Chinese and English literature on the treatment of malignant tumors with Wenyang prescriptions in the past 20 years.Among them,aconite is used the most frequently,but its antitumor mechanism is still unclear.In this project,we use liquid-mass spectrometry,network pharmacology,metabolomics,intestinal flora 16S rRNA sequencing and other technologies to explore the material basis and mechanism of action of Aconite against NSCLC from the perspective of metabolic reprogramming.The aconite used in this topic were all its decoction pieces Heishunpian,and the whole paper called it as aconite or aconite pieces.The HPLC fingerprint of the water extract of Aconite was firstly established in this subject,calibrating the components of 9 common peaks,and identifying the components of 5 chromatographic peaks.Since most alkaloids have no UV absorption,the subject further uses UPLC-Q-TOF-MS to identify and analyze the components in the liquid medicine,and uses UPLC-TSQ-MS to determine the content of 12 alkaloids,which are water extracts of aconite Establish quality control methods,and conduct a preliminary analysis of the material basis of Aconite against NSCLC.Secondly,the A549 tumor-bearing nude mouse model was used to study the anti-NSCLC pharmacodynamics of aconite,and the results showed that aconite can significantly inhibit the tumor volume and tumor mass of tumor-bearing nude mice,with a p value of less than 0.01 and no significant effect on body weight.Compared with the positive drug cisplatin,Aconite has no obvious side effects.Network pharmacology has shown that 62 active ingredients in Aconite have significant effects on PI3K-Akt-mTOR pathway,HIF-1 pathway and central carbon metabolism pathway through 29 core targets.All three pathways are related to the tricarboxylic acid(TCA)cycle.Related,we speculate that Aconite may play an anti-tumor effect by regulating the TCA cycle.Then nude mice feces were used to carry out microbial 16S rRNA sequencing.Through the analysis of intestinal flora diversity,community composition and 16S function prediction in nude mice,it is found that after modeling,the diversity of intestinal flora in nude mice is significantly reduced.The abundance of pathogenic bacteria increases and the abundance of probiotics decreases.Aconite can significantly improve the imbalance of intestinal flora,increase the diversity of the community,and increase the abundance of probiotics,especially Laospirillaceae.At the same time,it is found that Aconite can regulate the intestinal tract The abundance of the flora affects the anaerobic branching pathway of pentose phosphate.The results showed that Aconite can improve the intestinal flora and exert anti-tumor effects by regulating glycolytic pathways.Further metabonomics technology was used to study the mechanism of aconite inhibiting tumor growth in NSCLC tumor-bearing nude mice.The results of the non-targeted metabolomics test of tumor tissues indicate that Aconite may exert anti-tumor effcts through the branching glyoxylic acid and dicarboxylic acid cycles of the TCA cycle;the results of serum non-targeted metabonomics indicate that Aconite relies on the regulation of the TCA circulation pathway to achieve anti-tumor effects Effect:Serum targeted metabolomics results verify that Aconite can inhibit the growth of tumors by reducing the production of pyruvate,citric acid,α-ketoglutarate,succinic acid,fumaric acid,and malic acid,which are metabolites of the TCA cycle.Finally,the drug-containing serum of aconite was used to study its effect on the proliferation of NSCLC cells and its mechanism.The UPLC-Q-TOF-MS and UPLC-TSQ-MS technologies were used to qualitatively and quantify the relevant components in the medicinal serum of Aconite,combined with the preliminary network pharmacology prediction results show that the antiNSCLC material basis of Aconite is mostly monoester alkaloids and alcohol amines.Class ingredients.Tumor cell test results show that aconite medicated serum can significantly inhibit the proliferation of NSCLC cells,reduce mitochondrial membrane potential,increase intracellular ROS,induce apoptosis,and can also inhibit the expression of mTOR and PKM2 genes,and affect the upstream pathways of the TCA cycle.So as to play an anti-tumor effect.In summary,the monoester alkaloids and alcoholamine alkaloids in aconite may affect the mTOR signaling pathway,inhibit the TCA cycle metabolites,increase the abundance of the Laospirillaceae flora,regulate the glycolytic pathway,and induce tumor cell apoptosis.Die,exert anti-NSCLC drug effect.This project is the first to explore the material basis and mechanism of anti-NSCLC of Aconite from the perspective of metabolic reprogramming,providing a new scientific basis for the clinical treatment of NSCLC by Aconite,and providing new ideas for the anti-tumor research of warm-yang drugs.