Abstract:ObjectiveTo observe the influence of Jianpi Huoxue Qushi Decoction on nude mice bearing H22 hepatoma,and to investigate the possible mechanism of Jianpi Huoxue Qushi Decoction by interfering with Aquaporin 9 (AQP-9) protein target to regulate mitochondrial apoptosis pathway in hepatoma cells.MethodsH22 hepatoma cells were used to establish subcutaneous xenograft models,25 nude mice were selected.All the nude mice bearing H22 were randomly divided into blank group (normal saline gavage,0.2 ml/10 g),positive control group (cyclophosphamide intraperitoneal injection,30 mg/kg) and low-,medium- and high-dose groups (Jianpi Huoxue Qushi Decoction intragastric administration,25,50,100 mg/kg,respectively),with 5 rats in each group.nude mice bearing H22 were intervened once a day.The inhibition rate and spleen index of nude mice bearing H22 in each group were calculated.Fluorescent probe JC-1 and flow cytometry were used to detect the expression of mitochondrial membrane potential (ΔΨm).Western blot was used to detect the expression of B-cell lymphoma/leukemia-2 (Bcl-2) and B-lymphocyte tumor factor associated X protein (Bax).Immunohistochemistry was used to detect the expression of AQP-9.ResultsCompared with the blank group,the survival status of nude mice in the positive control group was worse,and the tumor volume,the spleen index and ΔΨm were significantly reduced,the tumor inhibition rate,the expression of Bcl-2 and Bax protein were significantly increased in the positive control group (P<0.05,P<0.01).Immunohistochemical results showed that the expression of AQP9 was increased in the positive control group.Compared with the positive control group,the tumor volume in the high-dose group significantly reduced,without significant chang in the tumor weight and tumor inhibition rate (P>0.05),the spleen index increased (P<0.05),the ΔΨm significantly decreased (P<0.05),the expression of Bcl-2 and Bax and the ratio of Bax/Bcl-2 significantly increased (P<0.05).The results of immunohistochemistry showed that AQP9 was widely expressed and stained dark brown.Some cells showed morphological changes of nucleus and increased cell vacuoles.ConclusionsJianpi Huoxue Qushi Decoction has obvious antitumor effect on nude mice bearing H22 hepatoma,and can improve the immune function of nude mice.The effect may be related to the regulation of AQP-9 intervention on mitochondrial apoptosis pathway of hepatoma cells.