The Research On The Mechanism Of Shuhuiwan Inhibiting HIF-1? To Improve The Microenvironment Hypoxia And Immune Escape Of Liver Cancer

Objective: This study starts from two aspects of improving tumor microenvironment hypoxia and immune escape,to investigate the effect and mechanism of Shuyuwan Decoction on the treatment of hepatocellular carcinoma.Methods: The subcutaneous liver cancer model was established,Male BALB/c-nu/nu mice(n=24,5 weeks old),mice were randomly divided into the following four groups(n=7 per group): Model group(administered 0.9%normal saline 0.2ml/d),S group(0.4g/d Shuyuwan Decoction),S+H group(0.4g/d Shuyuwan Decoction + weekly intraperitoneal dose of 5mg/kg DDP),H group(weekly intraperitoneal dose of 5mg/kg DDP).The gavage dose of each nude mouse was 0.2ml/d.The tumor volume was measured every two days,and the subcutaneous tumor was removed after 14 days of continuous intervention.The mitochondrial structure and number of tumor tissues were observed under electron microscope,and the mitochondrial proteins were quantitatively detected;Western Blot was used to detect the protein expression of HIF-1?,p53,PD-1,PD-L1,CD4 and CD8 in subcutaneous transplanted tumors of nude mice of human liver cancer,and RT-q PCR was used to detect PD-1,PD-L1,HIF-1? and p53 in real-time.The expression of CD4 and CD8 cells was observed by immunofluorescence method.Results: Compared with the model group,Shuyuwan Decoction could inhibit the growth of subcutaneous transplanted tumors in nude mice of human liver cancer,increase the amount of mitochondrial protein and protect the mitochondrial structure.At the same time,the expression of the oncogene HIF-1? and the negative immune regulatory factor PD-1/PD-L1 were inhibited,and the expression of the tumor suppressor gene p53,immune lymphocyte CD4 and CD8 were up-regulated,and the effect was enhanced after combined with chemotherapy.Meanwhile,HIF-1? was positively correlated with the expression of PD-1/PD-L1,and negatively correlated with the expression of CD4 and CD8.Conclusion: Shuyuwan Decoction can inhibit the growth of human hepatocellular carcinoma subcutaneously in nude mice.By improving the chemotherapy resistance caused by hypoxia in tumor microenvironment,the anti-tumor effect is significantly enhanced when used in combination with chemotherapy.On the one hand,its mechanism of action is to promote the inactivation of HIF-1? and the activation of p53,protect the mitochondrial structure and increase its number,and improve the glycolytic metabolism caused by hypoxia in the tumor microenvironment.On the other hand,the inhibition of negative immune regulators PD-1 and PD-L1 is mediated through the HIF-1? signaling pathway,and promoted the expression of CD4 and CD8 in tumor lymphocytes which can killing tumors,to avoid the occurrence of immune escape of tumor cells.Therefore,by inhibiting HIF-1? signaling pathway,Shuyuwan Decoction could improve the hypoxia and immune escape of hepatocellular carcinoma.