| Objective Liver cancer is one of the important causes of cancer death.Chinese medicine has obvious advantages in the treatment of primary liver cancer.To provide new ideas and methods for the diagnosis and treatment of liver cancer in traditional Chinese medicine,and to explore the effectiveness of traditional Chinese medicine in the treatment of liver cancer by poisoning phlegm and stasis spleen deficiency.Of outpatients with primary liver cancer conducted data mining,and conducted CCK-8 experiments to summarize the medication rules and compound prescriptions in the diagnosis and treatment of primary liver cancer;From the Wnt signaling pathway,the mechanism of Dutanyududutanyupi deficiency formula in promoting liver cancer cell apoptosis and inhibiting migration and invasion of liver cancer cells was clarified.Methods Part I: Collect the primary liver cancer patient data of Professor Wu Shoushan and Professor Sheng Guoguang,extract,cleanse and convert the data,and establish a database of Chinese medicine prescriptions in Excel 2019.The statistical method uses SPSS 26.0 software,uses description analysis to perform frequency statistics on traditional Chinese medicine prescription data,and uses systematic clustering method to perform cluster analysis on traditional Chinese medicine prescription data.Using IBM SPSS Modeler 14.1 analysis software,Apriori algorithm was used to analyze association rules of traditional Chinese medicine prescription data,and clarified the rules of Wu Shoushan and Sheng Guoguang’s diagnosis and treatment of liver cancer.CCK-8 cytotoxicity experiment was conducted on 23 single Chinese herbs for the treatment of liver cancer,and the Chinese herbal compound for the treatment of primary liver cancer was discussed.Part II: Compare the effects of Dutanyufang and Dutanyupifang on apoptosis,cell migration and cell invasion in human hepatocellular carcinoma cells,as well as β-catenin,c-Myc,Cyclin D1 and GSK in cells-3β and MMP2 genes were detected by real-time fluorescence quantitative PCR,which clarified the scientific basis for the treatment of primary liver cancer with Dutanyuyupi prescription.Statistical methods were analyzed using SPSS 26.0 software.All data were expressed as mean ± standard deviation(`x±s),comparison among multiple groups was performed by single factor analysis of variance,and comparison between groups was performed by LSD test.All the statistical tests used the two-sided test,P<0.05 was considered statistically significant.Result Part I: A total of 180 patients with primary liver cancer were included in the study.Poria,Yinchen,Licorice,Salvia miltiorrhiza,Radix Paeoniae Rubra,and Xiangfu had the highest frequency of drug use and the strongest association rule analysis.Cluster analysis classifies Chinese medicine into 8 categories.The two professors reused spleen-invigorating Chinese medicines in the diagnosis and treatment of primary liver cancer.The treatments often used spleen-invigorating and qi-invigorating,heat-clearing and detoxifying,dampening phlegm,nourishing yin and nourishing kidneys,and qi and stagnation.The CCK-8 experiment on a single Chinese herbal medicine,according to the proliferation rate from low to high,they are: Phyllanthus urinaria,Bindweed flower,Saponaria officinalis,Scutellaria barbata,Salvia miltiorrhiza,Hedyotis diffusa,which has a great influence on the proliferation rate of H22 cells.The single Chinese herbs are mainly detoxification,phlegm and blood circulation drugs.Part II: In the apoptosis experiment,both Dutanyufang group and Dutanyuyupi group can induce apoptosis of liver cancer cells,but Dutanyuyupi group is better than Dutanyu group.In the cell migration and invasion experiments,it was observed that the two Chinese herbal compound can inhibit the invasion and migration of liver cancer cells,and the effect of Dutanyuyupi group was better than Dutanyufang group.In the PCR experiment,both the Dutanyuyufang group and the Dutanyuyupi group reduced the gene expression of β-catenin,c-Myc,Cyclin D1,GSK-3β,and MMP2,but the effect of the Dutanyuyupi group was better than Dutan Yufang group.The prescription of poisoning phlegm and spleen deficiency can promote the apoptosis of liver cancer cells and inhibit the migration and invasion of liver cancer cells by reducing the gene expression of β-catenin,c-Myc,Cyclin D1,GSK-3β,MMP2.The mechanism of Fang’s anti-hepatoma may be related to blocking Wnt signaling pathway.Conclusion 1.According to the theory of poisoning phlegm and blood stasis,the data of patients with primary liver cancer in Professor Sheng Guoguang and Professor Wu Shoushan’s outpatient clinics from 2016 to 2017 were found.Poria,Yinchen,licorice,salvia,red peony,and Xiangfu were treated as primary treatments.Commonly used drugs for liver cancer,and the combination of Liuwei Chinese medicine is the most relevant.The two professors treat primary liver cancer and reuse spleen-invigorating Chinese medicine.Starting from the cytotoxicity of the CCK-8 experiment,the single Chinese traditional medicine for the treatment of liver cancer was optimized,and it was found that Phyllanthus indicus,Convolvulus,Saponaria,Scutellaria barbata,Salvia miltiorrhiza,and Hedyotis diffusa could effectively inhibit the proliferation of liver cancer cells.2.Through genomics research,it was found that the prescription of Dutanyuyupi can effectively promote the apoptosis of liver cancer cells,inhibit the migration and invasion of liver cancer cells,and its mechanism of action is mainly related to the Wnt signaling pathway. |
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