化癥回生丹及其拆方对Lewis肺癌模型小鼠IL-4、IL-12及Tregs细胞的影响

Effect of Huazheng Huisheng Dan and its disassembled prescription on IL-4,IL-12and Tregs cells in Lewis Lung Cancer Model mice

目的:探讨化癥回生丹及其拆方影响肿瘤免疫微环境的作用机制。方法:将112只C57BL/6小鼠按照体重随机分为化癥回生丹全方组、活血化瘀组、软坚散结组、行气止痛组、补气补血组、模型组、顺铂组及空白组,每组12只。除空白组外,其余小鼠于臀背部构建Lewis小鼠肺癌模型。各中药组每日予以0.4mL各中药组汤剂(生药剂量为5.8g/kg)灌胃,模型组及空白组每日予以等量生理盐水灌胃,顺铂组将顺铂注射液用生理盐水稀释至0.002g/kg,每只小鼠腹腔注射0.4mL,每周两次,并且每天予以等量生理盐水灌胃。14d后,处死小鼠,取肿瘤组织计算抑瘤率;取内眦静脉血,ELISA法检测IL-4,IL-12表达,流式细胞技术检测CD4^(+)Tregs、CD4^(+)CD25^(+)Foxp3^(+)Tregs表达。结果:与模型组比较,全方组、活血化瘀组、软坚散结组、补气补血组肺癌小鼠外周血IL-4水平明显降低,数据差异具有统计学意义(P<0.05);与模型组比较,化癥回生丹全方组、活血化瘀组、软坚散结组、补气补血组肺癌小鼠外周血IL-12水平明显回升,高于模型组IL-12水平,数据差异具有统计学意义(P<0.05);与模型组比较,化癥回生丹全方组、活血化瘀组、软坚散结组、补气补血组可显著降低外周血中CD4^(+)CD25^(+)Foxp3^(+)Tregs/CD4^(+)Tregs细胞比例,差异具有统计学意义(P<0.05)。结论:化癥回生丹及其拆方可能通过下调IL-4表达,促进IL-12分泌增加,降低CD4^(+)CD25^(+)Foxp3^(+)Tregs细胞比例,对肿瘤免疫微环境产生影响。

Objective:To study the mechanism of anti-tumor immune micro-environment.Method:According to their body weight,112C57BL/6mice were randomly divided into three groups:Huazheng Huishengdan group,Huoxue Huayu group,Ruanjian Sanjie group,qi analgesia group,tonifying qi and tonifying blood group,model group,cisplatin group and blank group with 12mice in each group.Except for the blank group,the lung cancer model of Lewis mice was established in the gluteal back of the other mice.Each traditional Chinese medicine group was given 0.4ml decoction(5.8g/kg)every day,the model group and blank group were given the same amount of saline every day,and the DDP injection was diluted to 0.002g·kg^(-1) in the platinum group,and 0.4ml was injected intraperitoneally in each mouse.After 14days,the mice were killed and the tumor tissue was taken to calculate the tumor inhibition rate.The expression of IL-4,IL-12was detected by ELISA method and the expression of CD4Tregs,CD4CD25Foxp3Tregs was detected by flow cytometry.Results:Compared with the model group,the levels of IL-4in peripheral blood of lung cancer mice in the whole prescription group,Huoxue Huayu group,Ruangjian Sanjie group and tonifying qi and tonifying blood circulation group were significantly lower than those in the model group(P<0.05).Compared with the model group,Huazheng Huishengdan whole prescription group,Huoxue Huayu group The level of IL-12in peripheral blood of lung cancer mice in Xuehuayu group,Ruangjian Sanjie group and invigorating qi and tonifying blood group was significantly higher than that in model group,and the differences in data were statistically significant(P<0.05).Compared with the model group,Huazheng Huisheng Dan group,Huoxue Huayu group,Ruanjian Sanjie group and tonifying qi and tonifying blood group could significantly reduce the proportion of CD4CD25Foxp3Tregs/CD4Tregs cells in peripheral blood,the difference was statistically significant(P<0.05).Conclusion:Huazheng Huisheng Dan and its disassembled prescription may promote the secretion of IL-12by down-regulating the expression of IL-4.Increasing and decreasing the proportion of CD4CD25Foxp3Tregs cells has an effect on tumor immune microenvironment.