Study On The Inhibition And Mechanism Of Sanghuang Extract On Subcutaneous Transplanted Tumor Of Cervical Cancer In Mice

Phellinuslinteus is a traditional Chinese medicine for the treatment of fungal diseases.Due to its numerous pharmacological components,such as polysaccharides,flavonoids,pyranones,and various triterpenoid compounds,it has great application prospects in the field of tumor treatment.According to the different extraction methods,the extracts of Sanghuang are also different,mainly divided into alcohol extract and water extract.Among them,yellow berry polysaccharides are the main components extracted from water,while alcohol is a derivative mainly containing flavonoids and trisaccharides.A large number of scholars have found that Sanghuang shows extraordinary potential in inhibiting tumor growth,regulating immunity,and inhibiting body oxidation.It can achieve anticancer therapeutic effects by stimulating host immunity and preventing cancer cell metastasis,while reducing the side effects of other anticancer drugs.Cervical cancer is one of the malignant tumors that have long plagued women around the world,and women with cervical cancer are more likely to die.which is a severe test for women ’s healthy life and life safety.Today,cervical cancer remains one of the most important public health issues in the world.Many traditional Chinese medicines have been widely used in the field of anti-cancer,but there are few reports on the anti-cervical cancer effect of Sanghuang in vivo,and its mechanism is still unclear.In this paper,Objective: To establish a cervical cancer tumor mouse model by subcutaneous injection of Kunming female mouse U14 cervical cancer cells,and to normalize its immune function.After the completion of subcutaneous injection,from the second day on a daily basis for Sanghuang extract(dose from high to low : 400 mg / kg,400 mg / kg and 200 mg / kg)gavage,Bajo el control del grupo,en el muestreo físico realizado en el área de vía,hay programas de audio o televisión claros.No hay un modelo de apoyo financiero,siempre que haya una estructura organizativa estable,no hay una tendencia a obstaculizar la movilidad de la población.The anti-cancer effect of Sanghuang extract was compared and analyzed by calculating the tumor inhibition rate.The safety of Sanghuang extract was analyzed by analyzing the changes of body weight and tissue structure of mice.At the same time,Illumina Hiseq 4000 high-throughput RNA-seq sequencing technology was used to perform transcriptome-level sequencing and bioinformatics-related analysis on the tumor tissues of mice in the intervention group and the model group of Phellinus igniarius extract,to screen out differentially expressed genes,in-depth bioinformatics discussion,and initially clarify the inhibitory mechanism of Phellinus igniarius extract on cervical cancer.Meanwhile,a 16 second rDNA sequence was used to evaluate the changes in the number and structure of plants in the intestine of tumor bearing mice after intervention with yellow blueberries extracted from water.The main experimental results are as follows :(1)Study the effect of Huangraspberry extract on tumor growth in mice.The water extraction effect of the three light group is higher than that of the 5-FU group,and the anti-tumor effect of Sanghuang water extract medium dose was the best,reaching67.54 %.The weight of the control group and another group of mice was significantly lower than that of the model control group,which may be related to an increase in tumor weight.This is a staining of mouse tumor tissue,indicating that the nuclear cells of the intervention group mice are relatively low,a large number of cancer cells have disintegrated,and a large number of cancer cells cannot see their nuclei.Tumor necrosis zone.The results showed that the device treated the trade of mouse liver,spleen,and kidneys more closely to the blank group than the model of extracting alcohol from yellow berries,which means extracting water from yellow berries.It has a protective effect on mouse liver function,has no obvious toxic side effects on mouse kidneys,and has a protective effect on mouse spleen and immune function.(2)Transcriptome results showed that there were a large number of Differentially Expressed Genes(DEGs)between the tumor tissues of the control group and the tumor tissues of the middle dose group after treatment,which could be further analyzed for functional enrichment.The gene function of cervical cancer cells will change under the intervention of Sanghuang aqueous extract,mainly involving the expression of genes involved in cellular processes and metabolic processes.After treatment,the expression of key genes involved in autophagy pathway,such as GABARAP,VMP1,VAMP8 and STX17,was regulated.In addition,the expression of genes involved in insulin signaling pathway,such as SLC2a4(GLUT4),RS2 and PRKAA2(AMPK),The expression is also changing.Through analysis of the Numenon gene,we found that the mechanisms of action of these three types of light are related to autophagy,apoptosis,and insulin signaling pathways.The above results were also confirmed by testing the Western version of LC3Ⅱ/LC3Ⅰ.It was found that the middle dose group of Sanghuang aqueous extract had an increasing trend in autophagy of cervical cancer after intervention.RT-PCR verification showed that the gene expression levels related to insulin signaling pathway,autophagy and cell signaling pathway were consistent with the expression levels in the transcriptome.It was further pointed out that the anti-tumor effect of Sanghuang aqueous extract in this study was mainly through affecting the glycolysis process,resulting in autophagy and apoptosis of cancer cells.(3)By analyzing the DNA sequence of intestinal microbiota 16 S,it was demonstrated that extracting water from yellow berries can alter the composition of intestinal microbiota in cancer mice.Each group of intestinal bacteria is mainly composed of three types of bacteria,with bacteria being the most important,followed by bacteria,Firmicutes.The abundance of Akkermansia increases with the extraction of yellow blueberries with moderate doses of water.Akkermansia has attracted much attention due to its potential benefits in metabolic regulation and cancer immunotherapy.Some studies have found that Sanghuang polysaccharide can regulate insulin signaling pathway by modifying intestinal flora.In this study,the changes of intestinal flora in Verrucomycota provide a basis for Sanghuang polysaccharide to intervene in cancer treatment through intestinal flora.(4)By analyzing the correlation,we found an important relationship between gut microbiota and host metabolism,which is related to the body weight coefficient and tumor weight of mice,as well as genes related to autophagy metabolism in tumor tissue.Conclusion:In vivo studies revealed that Morus alba extract significantly inhibited the growth of cervical tumours in U14 tumour-bearing mice by inducing tumour autophagy.The transcriptomic results suggest that the mechanism may be related to the effect of Morus alba extract on tumour metabolic reprogramming,particularly the inhibition of insulin-mediated glucose uptake and utilisation,and that intestinal flora may mediate the disturbance of glucose metabolism in tumours.