Study On The Mechanism Of Xiayuxue Decoction In Regulating ROS Level Through Cyclin-CDK Signaling Pathway To Prevent Liver Cance

OBJECTIVE: To detect the interventional effect of Xia Yu Xue decoction on liver cancer and to explore the potential efficacy and target mechanism of Xia Yu Xue decoction against liver cancer.METHODS: A subcutaneous transplantation tumor model of liver cancer in nude mice was established,and the content of the main components of Xia Yu Xue decoction was determined by high performance liquid chromatography.At 14 days of gavage,the nude mice were executed and weighed,and the pathological changes were observed by HE staining;apoptosis in the tissues was detected by TUNEL;the level of ROS in the tissues was detected by the kit probe,and the expression of Nrf2 in the nuclei of the tumor tissues was detected by immunofluorescence to analyze the alteration of the antioxidant system program in the tumor tissues;RT-q PCR and Western Blot were used to predict the bioinformatics of the intervention.The targets CHEK1,CDK1,CDK4,cyclin B1,cyclin B2,cyclin E1 transcripts and protein expression levels.RESULTS: Significant decrease in tumor volume(P < 0.05),decrease in tumor weight(P <0.05),and increase in apoptosis in subcutaneous transplantation tumor experiments in nude mice with hepatocellular carcinoma intervened with Xia Yu Xue decoction;accompanied by elevated ROS in tumor tissues(P < 0.01)and decreased levels of antioxidant defense protein Nrf2 into the nucleus;bioinformatically predicted targets of intervention,the LXYXD groups CHEK1(P <0.05),cyclin B1(P < 0.05),cyclin B2(P < 0.05),cyclin E1(P < 0.05),CDK1(P < 0.05),CDK4(P< 0.05)transcripts were significantly reduced in the LXYXD group,and in the HXYXD group CHEK1(P < 0.05),cyclin B1(P < 0.05),cyclin B2(P < 0.05),cyclin E1(P < 0.05),CDK1(P <0.05),CDK4(P < 0.05)transcripts were significantly reduced in the LXYXD group,and the expression of CHEK1(P < 0.001),CDK1(P < 0.001),CDK4(P < 0.05),cyclin B2(P < 0.05)proteins were reduced in the LXYXD group.In the HXYXD group,CHEK1(P < 0.001),CDK1(P < 0.001),CDK4(P < 0.05),and cyclin B2(P < 0.05)protein expression was reduced,and cyclin B1 and cyclin E1 protein expression did not show statistically different changes.CONCLUSION: In this study,combining bioinformatics and high performance liquid chromatography,animal experiments showed that the intervention of Xia Yu Xue decoction in hepatocellular carcinoma produced an inhibitory effect on the proliferation of Hep G2 hepatocellular carcinoma cells,increased apoptosis,accompanied by an increase in ROS levels in hepatocellular carcinoma cells,as well as a decrease in Nrf2 entry into the nucleus,and inhibition of CHEK1 and cyclin-CDK cell proliferation signaling pathways.