双歧杆菌DNA对小鼠腹腔巨噬细胞分泌和杀瘤功能的影响

Effect of Bifidobacterium DNA on Cytokine Production and Cytotoxicity in Peritoneal Elicit Macrophage of Mice

[目的]探讨双歧杆菌DNA对小鼠腹腔巨噬细胞的激活作用。[方法]纯化提取双歧杆菌基因组DNA ,并鉴定DNA制品CpG基序甲基化程度。收集双歧杆菌DNA(10μg/ml)体外诱导培养24h的小鼠腹腔巨噬细胞培养上清 ,采用ELISA法检测培养上清中IL 1β、IL 6的含量 ,用Griess试剂检测其NO的含量 ;采用MTT间接法观察了双歧杆菌DNA诱导的巨噬细胞体外杀伤活性的变化。[结果]实验组(双歧杆菌DNA组)巨噬细胞产生IL 1β、IL 6及NO的水平分别为(270.12±31.51)pg/ml,(578.81±47.30)pg/ml和(11.12±1.01)μmol/L,其巨噬细胞杀伤活性为(31.02±1.91)% ;对照组(PBS组)分别为(138.12±9.24)pg/ml、(78.80±10.21)pg/ml、(2.14±0.82)μmol/L和(18.69±1.70) % ,两组间各项指标比较均有显著性差异(均P<0.01)。[结论]双歧杆菌DNA能激活巨噬细胞 ,可增强巨噬细胞IL 1β、IL

To explore the active effect of Bifidobacterium DNA on peritoneal Elicit macrophages in mice(PEM Φ). The DNA of bifidobacteria was purified, and the degree of unmethylated CG dinucleotides (CpG motifs) was tested. The culture supernatant of PEM Φ which had exposed to Bifidobacteria DNA(10μg/ml) for 24hr was collected, and then the IL-1β, IL-6 were detected by ELISA method and the content of nitric oxide(NO)was explored by Griess reagent. The cytotoxicity of the PEM Φ was detected by indirect method of MTT. The levels of IL-1β, IL-6 , NO and the cytotoxicity rate of the PEM Φ in the experimental group (bifidobacteria DNA group) were (270.12±31.51)pg/ml,(578.81±47.30)pg/ml,(11.12±1.01)μmol/L and (31.02±1.91)% respectively, and those in the control group (PBS group) were (138.12±9.24) pg/ml,(78.80±10.21) pg/ml,(2.14±0.82) μmol /L and (18.69±1.70)% respectively. The differences were statisfically significant(P<0.01).[Conclusion]Bifidobacteria DNA can activate PEM Φ to secrete a lot of IL-1β, IL-6 , NO and can enhance its cytotoxicity.