Study On The Anti-tumor Efficacy And Mechanism Of Bruceajavanica Oil Emulsion

In 2022,the national cancer statistics released by the National Cancer Center showed that malignant tumor deaths account for 24.09% of all causes of death among residents.Cancer incidence and mortality have been on the rise in the past decade,and the annual medical expenses due to malignant tumors exceed 220 billion yuan,Bruceajavanica oil emulsion(BJOE)is a new and targeted pure Chinese herbal anti-cancer preparation with definite efficacy and low toxic side effects.It is a widely used anti-tumor traditional Chinese medicine in clinical practice and has more advantages over commonly used chemotherapy drugs in improving the quality of life and prolonging the survival of patients with advanced cancer.This product has the functions of inhibiting the proliferation of cancer cells and enhancing the immunity of the body.It is mainly used clinically in the treatment of lung cancer,digestive tract cancer,and melanoma.However,due to the lack of studies on its mechanism of action,especially the scientific elaboration on the mechanism of action of its combination,it is urgent to conduct in-depth studies on its anti-tumor efficacy and mechanism of action,especially the evaluation of the efficacy of its combination with anti-PD-1,for better guiding clinical application.The main contents and results of this study are as follows:The material basis,functional target,pathway,and molecular mechanism of melanoma prevention and treatment were discussed by network pharmacology.15 active components were selected,251 target genes were predicted,1475 disease targets were obtained,and 85 potential targets to inhibit melanoma were identified.Key genes were obtained,including AKT1,EGFR,JUN,etc.GO analysis mainly enriched signal transduction,protein phosphorylation,ATP binding,and protein kinase activity,KEGG pathway analysis indicated that pathways in cancer,Endocrine resistance,VEGF signaling pathway,PI3K-Akt signaling pathway,and IL-17 signaling pathway,and other related pathways were involved.The improvement of tumor microenvironment(TME)and the enhancement of anti-tumor effect in combination with anti-PD-1 of BJOE was investigated.the results of MTT assay showed that the anti-tumor mechanism of BJOE may not be the direct killing of tumor cells in the B16 and CT26 tumor animal model.The effect of BJOE on improving the TME and on immune organs was further explored in the B16 tumor animal model,and the results indicated that BJOE could down-regulate the IL-10 level in the TME.When combined with anti-PD-1,the experimental animals showed a higher survival rate and better tumor growth inhibition effect than the saline group.Compared with the saline group,the number of CD3(+)cytotoxic T lymphocytes in both spleen and lymph nodes of mice increased after treatment with PD-1and BJOE,respectively,and this effect was more significant in the anti-PD-1 combined with BJOE treatment group.Also,in mouse lymph nodes,it was found that BJOE could promote the proliferation of helper/induced CD4+ T lymphocytes.In the CT26 tumor mouse model,the combined drug group was comparable to the BJOE group and anti-PD-1 group in terms of tumor growth inhibition,and its difference with the saline group was gradually significant after 15 days of treatment.This suggested that BJOE has certain anti-tumor activity in the CT26 tumor model.In terms of survival rate improvement,the BJOE alone and anti-PD-1 combined with BJOE treatment groups had comparable effects,both better than the anti-PD-1 treated and saline groups.In the 4T1 tumor mouse model,it was found that BJOE had good biosafety,and BJOE alone also had an anti-tumor effect.CD31+ staining results showed that BJOE had a significant inhibitory effect on angiogenesis.H&E staining of tumor showed that the tumor tissue in the BJOE-treated group was vacuolated,indicating that its structure was destroyed.Those results revealed that BJOE could achieve anti-tumor effects through anti-angiogenesis.A FRET system was constructed to predict the release behavior of BJOE in vivo,and the results showed that it was mainly released at tumor sites.Using LC-MS/MS method and oleic acid as the detection index,it was found that the main distribution organs of BJOE were: stomach > lung > intestine > fat > liver > ovary >muscle > spleen.and the distribution concentrations and trends of BJOE in the three batches from different manufacturers were similar at 0.5h,2h,and 6h,with no significant difference by statistical analysis.Accelerated(0,1,2,3,6 months)stability investigation was carried out on 3 batches of BJOE according to the proposed quality standards,including hemolysis and coagulation test,antihypertensive substance test,allergy test,pyrogen test,and abnormal toxicity test.All the test results were in line with the regulations.