| Background: hepatocellular carcinoma(HCC)is a common malignancy with nearly 900,000 new cases of liver cancer and 830,000 deaths due to liver cancer annually worldwide,and liver cancer has seriously affected people’s living standards and health.China is a country with a high incidence of liver cancer,and the number of patients with liver cancer accounts for approximately 55% of the total liver cancer population worldwide.The early symptoms of the onset of liver cancer are insidious and difficult to detect,and advanced liver cancer is difficult to cure by surgical resection,causing severe mental and economic pressure on the patient’s family and society.Therefore,it is vital to find effective means of combating liver cancer.Clinical results found that HCC patients are often accompanied by the deactivation of the body’s immune function.The body’s immune system is composed of immune organs,immune cells,and immune molecules.T cells are the primary anti-tumor immune executive cells,which play the role of directly killing tumor or indirectly killing tumor cells by secreting immune molecules through specifically recognizing tumor antigens,developing into effector T cells.Therefore,maintaining the stability of T cell antitumor immune function is particularly important for the body to exert antitumor responses.Traditional Chinese medicine(TCM)has a long history of application and a significant therapeutic effect on diseases.Early TCM had shown some understanding of liver cancer symptoms,and many Chinese ancient books had detailed descriptions of liver cancer symptoms.Studies have shown that TCMs can exert anti-inflammatory,anti-viral,and tumor-killing effects by regulating the body’s immune system metabolism.Therefore,suitable anti-liver cancer drugs can be screened from TCM.Purpose: To observe the effect of combination of beauverine and cinnamaldehyde in the treatment of liver cancer and the therapeutic effect on the impairment of mitochondrial function in peripheral blood T cells induced by hepatocarcinoma,and to explore the specific mechanism of combination of beauverine and cinnamaldehyde in the treatment of liver cancer.Methods:(1)we first established a mouse liver cancer model by tail vein high-pressure injection of H22 liver cancer cells to examine the effects of different cell injection amounts on the preparation of a liver cancer model and the mouse immune system.(2)The mitochondrial damage model of T cells induced by benzene exposure was established to observe the repair effect of bassiamine and cinnamaldehyde on T cells with damaged mitochondria.Using a mouse model of liver cancer,the anti-hepatoma effects of beauverine and cinnamaldehyde and their effects on the mitochondrial function of peripheral blood T cells were investigated.Next,we obtained liver cancer-specific T cells by presenting tumor antigens by DC cells,and established a T cell and tumor cell co-culture model in vitro,which was used to further investigate the mode of action of beauverine combined with Cinnamaldehyde to enhance T cell antitumor.(3)We chose BALB/c nude mice to prepare a liver cancer model,in vitro beauverine combined with cinnamaldehyde pretreatment of T cells,and then T cell anti-liver cancer effect in mice was verified by T cell reinfusion.Meanwhile,the effects of beauverine and cinnamaldehyde on T cell apoptosis and mitochondrial morphology were evaluated by transmission electron microscopy.Finally,in vitro experiments were performed to explore the specific action pathways of Beauvericin and cinnamaldehyde in regulating mitochondrial function by regulating mitochondrial-related functional protein expression in T cells using ZLN-005,mdivi-1.Results:(1)A mouse model of hepatocellular carcinoma was prepared by injecting H22 hepatocellular carcinoma cells into caudal vein.At the same time,the tissue structure of thymus and spleen of liver cancer mice was significantly damaged,the proportion of late apoptotic cells in peripheral blood T cells increased,and the levels of mitochondrial membrane potential and ATP decreased significantly.(2)In vitro exposure of benzene established a T-cell mitochondrial damage model,and CCK-8 results found that Beauverine and cinnamaldehyde could promote the proliferation of T cells,elevate T-cell mitochondrial membrane potential and ATP levels,and reduce T-cell mitochondrial function damage.The results of cell co-culture showed that beauveria bassiana combined with cinnamaldehyde could effectively improve the ability of T cells to secrete immune factors perforin and granulocase B,and induce tumor cell apoptosis.(3)The results of T cell transfusion experiment in nude mice showed that beauveristin combined with cinnamaldehyde could increase the number of T cell infiltration in tumor tissue and significantly reduce the number of liver cancer nodules in mice with liver cancer.The results of transmission electron microscopy showed that the number of T cell mitochondria in hepatocellular carcinoma mice was decreased compared with that in normal mice.The morphology of T cell was dumbbell type and apoptosis occurred.The T cell mitochondria were restored to normal state after treatment with Beauversiacin combined with cinnamaldehyde.(4)The mitochondrial synthesis and division proteins of T cells were detected,and the expressions of PGC-1α,NRF1,NRF2 and TFAM in the combination treatment group were significantly increased compared with those in the liver cancer group,while the expressions of p-DRP1 and DNM2 were significantly decreased,and the apoptosis of T cells was decreased.In addition,pretreatment with PGC-1α activator ZLN-005 and DRP1 inhibitor mdivi-1 showed that increased PGC-1α expression inhibited p-DRP1 expression,both of which inhibited T cell apoptosis.Conclusions: in summary,we successfully constructed a BALB / c mouse model of liver cancer by tail vein injection of H22 liver cancer cells and found that the immune system of mice with liver cancer was damaged and the mitochondrial function of peripheral blood T cells was defective.Combination treatment with beauverine and cinnamaldehyde leads to the activation of PGC-1α/drp1 pathway regulates T cell mitochondrial synthesis and division processes,enhances T cell mitochondrial function,and inhibits T cell apoptosis in hepatoma-bearing mice to exert anti-hepatoma effects.In this study,we investigated the effect of beauverine combined with cinnamaldehyde on the anti-liver cancer and its immunomodulatory mechanism,which may provide important reference value for TCM combined treatment of liver cancer. |
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