摘要
目的从树突状细胞(DCs)的分子细胞水平初步探讨华蟾素(CBC)预防肿瘤的作用机制。方法将20只Wistar大鼠随机分成两组,华蟾素干预组和阴性对照组各10只,0.01%二乙基亚硝胺(DEN)饮水,供两组大鼠自由饮用造成肝癌模型,共连续饮12周后再更换为正常饮水至20周,同时从第1周对干预组大鼠进行腹腔注射华蟾素(每周2次,共20周)。阴性对照组给予同等剂量的0.9%氯化钠溶液。通过核磁共振(MRI)、大体观察肝脏病变;应用重组细胞因子诱导、培养两组大鼠骨髓来源的树突状细胞(DCs),并在倒置显微镜下观察DCs的形态特点;流式细胞术检测DCs表型;ELISA法检测两组大鼠血清IL-2、IL-10浓度。结果经MRI检测,两组大鼠肝脏T2加权像显示均有多个小的高密度结节。CBC干预组的结节数(84.3±9.3)与阴性对照组(121.7±15.6)相比,差异具有统计学意义(P<0.05)。诱导DCs后与阴性对照组相比,倒置显微镜下CBC干预组中诱导分化形成的DCs细胞较典型。经流式细胞术检测,CBC干预组中DCs表面的CD80、CD86,MHC-Ⅱ类的阳性表达率均有明显增高。与阴性对照组比较,CBC干预组的DCs表面标志物差异均有统计学意义(P<0.05)。ELISA检测结果显示,与阴性对照组相比,CBC干预组的大鼠血清IL-2和IL-10差异均有统计学意义(P均<0.05)。结论华蟾素通过上调DCs表面标志物的表达,促进DCs的成熟度,从而抑制肝癌生长。
Objective To investigate the role of cinobufacini （CBC） in anti tumor mechanism at the molecular level of dendritic ceils （DCs）. Methods Female Wistar rat model with hepatic cancer was induced by 0.01% diethylnitrosamine （DEN）. Twenty Wistar rats were randomly divided into 2 groups （CBC intervention group, negative control group） after 12 weeks of DEN treatment, and received CBC intraperitoneal injection （2 times/week × 20 weeks）in intervention group or equal dosage of 0. 9% saline in control group, respectively. Livers were checked at the time of 20th week by 3.0 T MRI. Using recombinant cytokine to induce and culture bone marrow derived DCs and observe the morphology characteristics under the inverted microscope. Flow cytometry was used to detect the phenotypes of DCs. ELISA was applied to test the levels of serum IL- 2,IL- 10. Results MRI showed that the number of tumor nodules in intervention group （84.3 ± 9.3） was significantly less than that of in control group （121.7 ± 15.6）（P〈0.05）. Compared with negative control, the morphology of DCs in CBC intervention group was more typically under the inverted microscope. Moreover, the positive expressions of phenotypes ERT IB （MHC-II ）, CD80,CD86] were increased in CBC intervention group. There were significantly differences in pheno types between 2 groups（P〈0.05）, ELISA data showed that the levels of serum IL- 2,IL-10 were signifi cantly differences in CBC intervention group, compared with negative group（P〈0.05）. Conclusion CBC can up-regulate the expressions of DCs phenotypes, promote the mature of DCs, and enhance the secretion of Th1 cytokines to suppress the growth of liver cancer.
出处
《肿瘤防治研究》
CAS
CSCD
北大核心
2012年第10期1183-1187,共5页
Cancer Research on Prevention and Treatment
基金
乌鲁木齐市科技局资助项目(Y111310030)
关键词
华蟾素
肝癌
树突状细胞
大鼠
预防
Cinobufacini
Liver cancer
Dendritic cells
Rat
Prevent
