二烯丙基二硫化物诱导MDCC-MSB-1细胞的凋亡及Fas/FasL信号通路调控作用  

Diallyl Disulfide-induced Apoptosis of MDCC-MSB-1 Cells and Regulation of Fas/FasL Signaling Pathway

作　　者：王宜平 周鹏飞 叶正钦 李真诚 唐清秀 纪春晓[1,2] WANG Yi-ping;ZHOU Peng-fei;YE Zheng-qin;LI Zhen-cheng;TANG Qing-xiu;JI Chun-xiao(College of Veterinary Medicine,Hunan Agricultural University,Changsha,Hunan,410128,China;Hunan Engineering Technology Research Center of Veterinary Drugs,Changsha,Hunan,410128,China)

机构地区：[1]湖南农业大学动物医学院,湖南长沙410128 [2]湖南省兽药工程技术研究中心,湖南长沙410128

出　　处：《动物医学进展》2021年第8期7-13,共7页Progress In Veterinary Medicine

基　　金：湖南省自然科学基金面上项目(2020JJ4350);湖南省教育厅优秀青年项目(19B251);国家自然科学基金项目(31402258)。

摘　　要：为探究二烯丙基二硫化物(DADS)对马立克氏病肿瘤细胞系(MDCC-MSB-1细胞)的增殖抑制作用及其信号通路的调控机制,以MDCC-MSB-1细胞作为研究对象,采用CCK-8法检测不同浓度DADS分别作用MDCC-MSB-1细胞24,48,72 h后对细胞增殖抑制的影响。用0、80、160、240μmol/LDADS预处理MDCC-MSB-1细胞24 h后,光学显微镜观察细胞形态变化;Hoechst 33342核酸染色法检测细胞形态变化;caspase-8活性试剂盒检测细胞中caspase-8蛋白酶活性;并用荧光定量PCR法检测Fas/FasL信号通路关键因子mRNA的转录水平。结果显示,40μmol/L〜240μmol/L的DADS抑制MDCC-MSB-1细胞的增殖呈时间剂量依赖性;随着浓度的升高,细胞形态逐渐出现细胞核核膜消失,染色体降解为多个片段并且边缘化,形成凋亡小体;240μmol/L的DADS可诱导MDCC-MSB-1细胞caspase-8蛋白酶活性及Fas、FADD、caspase-8、Bid和caspase-3mRNA的转录水平极显著升高(P<0.01)。结果表明,DADS可诱导MDCC-MSB-1细胞的凋亡,并活化其Fas/FasL死亡受体凋亡信号通路,可为鸡马立克氏病的治疗提供参考。In order to explore the inhibitory effect of diallyl disulfide(DADS) on the proliferation of Marek's disease tumor cell line(MDCC-MSB-1 cells) and its signal pathway regulation mechanism, this study used MDCC-MSB-1 cells as the study subjects, the CCK-8 method was used to detect the effects of different concentrations of DADS on the inhibition of cell proliferation after treating MDCC-MSB-1 cells for 24,48,72 h.After pretreatment of MDCC-MSB-1 cells with 0,80,160,240 μmol/L DADS for 24 h, the morphological changes of the cells were observed under an optical microscope;Hoechst 33342 nucleic acid staining method was used to detect the morphological changes of the cells;Caspase-8 activity kit was used to detect the cell Caspase-8 protease activity;fluorescence quantitative PCR method was used to detect the transcription level of the key factor mRNA of Fas/FasL signaling pathway.The results showed that 40 μmol/L-240 μmol/L DADS inhibited the proliferation of MDCC-MSB-1 cells in a time-dose-dependent manner;with the increase of the concentration, the cell morphology gradually disappeared, and the chromosomes were degraded into multiple fragment and marginalization to form apoptotic bodies;240 μmol/L DADS can induce Caspase-8 protease activity in MDCC-MSB-1 cells and the transcription levels of Fas, FADD,Caspase-8,Bid and Caspase-3 mRNA are extremely significant increase(P<0.01).The results showed that DADS can induce the apoptosis of MDCC-MSB-1 cells and activate the Fas/FasL death receptor apoptosis signaling pathway.It can provide reference for the treatment of chicken Marek's disease.

关 键 词：二烯丙基二硫化物 鸡马立克氏病 Fas/FasL信号通路 凋亡 

分 类 号：S852.3[农业科学—基础兽医学]