华蟾素辅助顺铂化疗肝癌移植瘤小鼠的抑瘤作用及对肠道菌群、外周血淋巴细胞亚型的影响

Antitumor effect of cinocobalamin-assisted cisplatin chemotherapy in mice with liver cancer transplantation tumor and its effect on intestinal flora and peripheral blood lymphocyte subtypes

目的研究华蟾素对肝癌移植瘤小鼠中对顺铂抗肿瘤作用的影响,并探讨其对顺铂治疗的肝癌小鼠肠道菌群及外周血淋巴细胞亚型的影响。方法45只6~8周龄,雌雄不限,Balb/c小鼠通过腋下注射H22肝癌细胞建立肝癌移植瘤小鼠模型,随机数字表法分为模型组、顺铂组和华蟾素+顺铂组3组,各15只。顺铂组小鼠给予腹腔注射顺铂治疗,华蟾素+顺铂组给予腹腔注射顺铂和华蟾素治疗,模型组腹腔注射等量溶剂。2周后,安乐死各组小鼠,分离各组小鼠肿瘤组织、肠道组织和外周血,比较各组小鼠肿瘤大小、肠道益生菌含量、肠道菌群丰度以及外周血淋巴细胞亚型。结果华蟾素+顺铂组小鼠经治疗后的抑瘤率为(56.38±12.91)%,显著高于顺铂组小鼠[(40.43±11.82)%],差异有统计学意义(P<0.05)。顺铂组和华蟾素+顺铂组小鼠肠道益生菌(双歧杆菌+乳酸杆菌)含量、菌群丰度Shannon指数和Chao指数均低于模型组小鼠,并且华蟾素+顺铂组小鼠显著高于顺铂组小鼠,差异均有统计学意义(P<0.05)。顺铂组和华蟾素+顺铂组小鼠肠道肠道屏蔽功能指标DAO、内毒素和细菌异位率均高于模型组小鼠,并且华蟾素+顺铂组小鼠显著低于顺铂组小鼠,差异均有统计学意义(P<0.05)。此外,顺铂组和华蟾素+顺铂组小鼠CD4^(+)和CD8^(+)细胞比例均显著低于模型组,而CD4^(+)/CD8^(+)值均显著高于模型组,差异均有统计学意义(P<0.05);华蟾素+顺铂组小鼠外周血CD4^(+)细胞比例均显著高于顺铂组小鼠,并且CD4^(+)/CD8^(+)值均显著低于顺铂组小鼠,差异均有统计学意义(P<0.05)。结论华蟾素可以在体内增强顺铂抗肝癌作用,并且可以有助于提高顺铂治疗肝癌小鼠肠道益生菌含量、肠道菌群丰度、肠道屏蔽功能以及外周血细胞免疫功能。

Objective To study the effect of cisplatin on the anti-tumor effect of cisplatin in mice with liver cancer transplanted tumors,and to explore its effect on the intestinal flora and peripheral blood lymphocyte subtypes of liver cancer mice treated with cisplatin.Methods Forty-five 6~8 weeks old,male or female,Balb/c mice were injected with H22 hepatocellular carcinoma cells into the axillary to establish a liver cancer xenograft mouse model.They were divided into three groups:model group,cisplatin group,and cininobufosin+cisplatin group according to the random number table,each with 15 mice.Mice in the cisplatin group were treated with intraperitoneal injection of cisplatin,the cininobufosin+cisplatin group were treated with cisplatin and cisplatin,and the model group was intraperitoneally injected with the same amount of solvent.Two weeks later,each group of mice was euthanized,and the tumor tissue,intestinal tissue and peripheral blood of each group of mice were separated to compare the tumor size,intestinal probiotic content,intestinal flora abundance and peripheral blood lymphocytes of each group of mice subtype.Results After treatment,the tumor inhibition rate of mice in the cinobufacin+cisplatin group was(56.38±12.91)%,which was significantly higher than that in the cisplatin group[(40.43±11.82)%],and the difference was statistically significant(P<0.05).The content of probiotics(Bifidobacterium+Lactobacillus),Shannon index and Chao index of the intestinal tract of mice in the cisplatin group and the cinobufosin+cisplatin group were lower than those in the model group,the above indexes of mice in the cinobufacin+cisplatin group were significantly higher than those in the cisplatin group,and the differences were statistically significant(P<0.05).The DAO,endotoxin,and bacterial ectopic rate of the intestinal shielding function indexes of the mice in the cisplatin group and the cinobufosin+cisplatin group were higher than those in the model group,and the above indexes of mice in the cinobufacin+cisplatin group were significantly lower than those in the cisplatin group,and the differences were statistically significant(P<0.05).In addition,the ratio of CD4^(+) and CD8^(+) cells in the cisplatin group and the cinobufosin+cisplatin group was significantly lower than that in the model group,while the CD4^(+)/CD8^(+) were significantly higher than that in the model group,the differences were statistically significant(P<0.05);the ratio of CD4^(+) cells in the peripheral blood of the cinobufosin+cisplatin group was significantly higher than that of the cisplatin group,and the CD4^(+)/CD8^(+) values were significantly lower than those of the cisplatin group,the differences were statistically significant(P<0.05).Conclusion Cinobufosin can enhance the anti-liver cancer effect of cisplatin in vivo,and can help increase the intestinal probiotic content,intestinal flora abundance,intestinal shielding function and peripheral blood cell immune function of cisplatin-treated liver cancer mice.