香草扶正合剂对H22荷瘤小鼠免疫功能及肿瘤组织HuR、BIRC3、NF-κB P65蛋白表达的影响OA

Objective:To investigate the effects of Xiangcao Fuzheng Mixture(XCFZ)on the immune function and the expression of HuR,BIRC3,and NF-κB P65 proteins in tumor tissues of H22 tumor-bearing mice.Methods:A mouse model of liver cancer with tumor implantation was established using the H22 liver cancer cell line through the transplantation method.The mice were randomly divided into model group,cisplatin group(DDP),the low,medium and high doses of XCFZ groups,the combined group,with 10 mice in each group.From the second day after modeling,the low,medium and high doses of XCFZ were 9.3,18.6 and 37.2 g/kg respectively;the cisplatin group was intraperitoneally injected with 5 mg/kg cisplatin solution,and the same amount of double distilled water was gavaged simultaneously.The combined group was given XCFZ medium dose gavage and cisplatin intraperitoneal injection simultaneously.The model group was given the same amount of 0.9%sodium chloride solution once a day for 14 days.The body weight,tumor weight,spleen and thymus weight of the mice were measured,and the tumor inhibition rate,spleen-thymus index were calculated.The tumor tissue was stained with HE;the cell structure of the tumor tissue was observed under transmission electron microscope;the number of CD3+,CD4+and CD8+positive cells was detected by flow cytometry and the CD4+/CD8+ratio was calculated;the expression levels of serum cytokines(INF-γ,TNF-α,IL-2,IL-10)in mice were detected by ELISA;RT-PCR and Western blot were used to detect the mRNA and protein expressions of human antigen R(HuR),baculoviral IAP repeat containing 3(BIRC3),nuclear transcription factor-κB(NF-κB)in tumor tissue.Results:Compared with the model group,the combination group had the highest tumor inhibition rate and was significantly better than the other groups.The body weight of the mice in the cisplatin group was the lightest after tumor removal,and the combination group was better than the cisplatin group alone.Compared with the cisplatin group,the body weight of the mice in the XCFZ groups at all doses was significantly improved(P<0.05 or P<0.01).Compared with the model group,the medium-dose XCFZ group had a better improvement in the spleen-thymus index than the other XCFZ groups(P<0.05),and the combination group had a better spleen-thymus index than the cisplatin group(P<0.05).In the model group,the tumor cells were distributed disorderly,with many mitotic and atypical tumor cells;in the XCFZ groups,mitosis and atypical cells were rare,with many isolated cells and intercellular cavities,and even nuclear pyknosis and extensive tumor cell necrosis;in the cisplatin group,many fragmented and pyknotic nuclei and intercellular cavities were observed,with many autophagosomes and a large number of necrotic cells.In the XCFZ groups at all doses,the cytoplasm was reduced and many vacuoles appeared in the cells.Compared with the cisplatin group,the CD3+and CD4+contents in the XCFZ groups at all doses and the combination group were increased(P<0.05).Compared with the model group,the CD4+/CD8+ratio in the XCFZ groups at all doses was significantly increased(P<0.05 or P<0.01),and it was significantly decreased in the cisplatin group(P<0.05).Compared with the model group,the expression of inflammatory factors in the medium-dose XCFZ group was the most significantly decreased(P<0.05),and the combination group significantly reduced the expression of inflammatory factors more than the other treatment groups(P<0.01).Compared with the cisplatin group,the combination group significantly improved and reduced the expression of INF-γ,TNF-α,IL-2,and IL-10(P<0.01).Compared with the cisplatin group,the expression of HuR,BIRC3,and NF-κB P65 mRNA in the combination group was decreased(P<0.05),and the expression in the other XCFZ groups at all doses was more significantly decreased(P<0.05 or P<0.01).Compared with the model group,the expression of HuR,BIRC3,and NF-κB P65 in the medium-dose XCFZ group and the cisplatin group was significantly decreased(P<0.05 or P<0.01).Compared with the cisplatin group,the combination group had a better inhibitory effect on the expression of HuR,BIRC3,and NF-κB P65(P<0.05 or P<0.01).Conclusion:Xiangcao Fuzheng mixture can effectively inhibit tumor growth in H22 liver cancer-bearing mice,improve the body weight and immune organ function of the mice,and enhance T lymphocyte-mediated anti-tumor immunity.The mechanism may be related to the inhibition of HuR,BIRC3 and NF-κB P65 protein expression.