消退素D1对原位肝癌的抗癌作用及其对Th1/Th2平衡的影响

The anticancer effect of melatonin D1 on in situ liver cancer and Th1/Th2 balance in mice

目的探讨消退素D1对原位肝癌小鼠的抗癌作用及其对Th1/Th2平衡的影响。方法60只6~8周龄SPF级ICR雄性小鼠,随机分为对照组、模型组、低剂量组和高剂量组,每组15只。通过将H22细胞接种于小鼠肝左叶构建原位肝癌小鼠模型,对照组仅注射等量生理盐水,低剂量和高剂量组小鼠在模型组的基础上,分别每天腹腔注射2.5 mg/kg和10 mg/kg的消退素D1,连续3周。收集并称量小鼠肝质量,全自动生化仪检测小鼠血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)和总胆红素(TBil)含量。免疫组化法检测肝癌组织Ki67表达,流式细胞术检测小鼠肝癌组织Th1和Th2表达。RT-PCR法检测小鼠肝癌组织T-bet和Gata-3 mRNA表达。Western blot法检测小鼠肝癌组织T-bet和Gata-3蛋白表达。ELISA法检测小鼠血清IL-2、干扰素-γ(IFN-γ)、IL-4和IL-10含量。结果与对照组比较,模型组小鼠肝癌组织体积、Ki67表达、血清ALT、AST和TBil的含量均明显升高(P<0.05),肝癌组织中Th1比例、T-bet蛋白和mRNA表达、血清IL-2和IFN-γ含量显著降低(P<0.05),Th2比例、Gata-3蛋白和mRNA表达、血清IL-4和IL-10含量明显升高(P<0.05)。与模型组相比,低剂量组和高剂量组小鼠肝癌组织体积、Ki67表达、血清ALT、AST和TBil的含量均明显降低(P<0.05),肝癌组织中Th1比例、T-bet蛋白和mRNA表达、血清IL-2和IFN-γ含量显著升高(P<0.05),Th2比例、Gata-3蛋白和mRNA表达、血清IL-4和IL-10含量明显降低(P<0.05)。结论消退素D1通过促进Th1/Th2向Th1分化,抑制肝癌的生长和转移。

To investigate the role of resolvin D1 in hepatoma mice and its influence on Th1/Th2 balance,60 ICR male mice were recruited and randomly divided into control group,model group,low-dose group and high-dose group,with 15 mice in each group.The mice in model group were inoculated with H22 cells under the left liver lobe;the mice in the low-dose and high-dose groups were intraperitoneally injected with 2.5 mg/kg and 10 mg/kg of resolvin D1 every day for 3 weeks.The liver of mice was collected and weighed,and the contents of serum alanine aminotransferase(ALT),aspartate aminotransferase(AST)and total bilirubin(TBil)in mice were detected by an automatic biochemical analyzer.The expression of Ki67 was detected by immunohistochemistry and the expression of Th1 and Th2 were detected by flow cytometry.RT-PCR was used to detect the expression of T-bet and Gata-3 mRNA in mouse liver cancer tissues.The expression of serum IL-2,interferon-γ(IFN-γ),IL-4 and IL-10 were detected by ELISA.Compared with the control group,the liver carcinoma volume,Ki67 expression,and the serum content of ALT,AST and TBil in the model group were significantly increased(P<0.05),while the proportion of Th1,the protein and mRNA expression of T-bet,and the serum content of IL-2 and IFN-γwere significantly decreased(P<0.05),and the proportion of Th2,the protein and mRNA expression of Gata-3,and the serum content of IL-4 and IL-10 were significantly increased(P<0.05).Compared with the model group,the liver carcinoma volume,the serum content of ALT,AST and TBil in the low-dose group and high-dose group were significantly decreased(P<0.05),the proportion of Th1,the protein and mRNA expression of T-bet,and the serum content of IL-2 and IFN-γwere significantly increased(P<0.05),while the proportion of Th2,the protein and mRNA expression of Gata-3,and the serum content of IL-4 and IL-10 were significantly decreased(P<0.05).In conclusion,Resolvin D1 inhibits the growth and metastasis of liver cancer by promoting Th1/Th2 to differentiate into Th1.