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复方苦芩对感染猪传染性胃肠炎病毒的PK-15细胞中Bcl-2/BAX mRNA转录的影响
  • ISSN号:0578-1752
  • 期刊名称:《中国农业科学》
  • 时间:0
  • 分类:S852.659.6[农业科学—基础兽医学;农业科学—兽医学;农业科学—畜牧兽医]
  • 作者机构:[1]西南大学荣昌校区兽医科学研究中心中药创新研究室,重庆402460
  • 相关基金:国家公益性行业(农业)科研专项(201303040-05)
中文摘要:

【目的】探讨中药复方苦芩对感染猪传染性胃肠炎病毒(transmissible gastroenteritis virus of swine, TGEV)的猪肾细胞(PK-15)中凋亡基因Bcl-2/BAX mRNA转录的影响。【方法】体外扩增TGEV,用TGEV感染PK-15细胞的模型进行体外试验研究,试验中设有复方苦芩组、空白组、模型组及黄芪多糖组,在测定了病毒半数感染细胞量(TCID50),复方苦芩及黄芪多糖药液对PK-15细胞的最大无毒浓度后,制作细胞爬片采用瑞士染色法染色观察PK-15细胞的形态变化;按试剂盒操作提取各试验组RNA并及时反转录为cDNA,采用实时荧光定量PCR法检测细胞中Bcl-2和BAX mRNA转录情况。【结果】与空白组相比,模型组的PK-15细胞发生了明显的细胞病变效应(cytopathic effect,CPE),细胞变细长,细胞间成网状,细胞核着色较浅,出现核浓缩、固缩、碎裂等现象;与模型组比,复方苦芩组与黄芪多糖组细胞病变得到改善,而复方苦芩组效果更加明显,大部分细胞核结构清晰,染色质着色均一,少部分细胞核变大或碎裂;与空白组相比,模型组Bcl-2 mRNA转录量减少(P<0.01),复方苦芩组Bcl-2基因转录量增加,为模型组的1.403倍(P<0.01)、黄芪多糖组的1.078倍(P>0.05);与空白组相比,模型组BAX mRNA转录量明显升高,而其他3组BAX基因转录差异不明显(P>0.05),模型组BAX基因转录量为空白组的1.440倍、复方苦芩组1.290倍、黄芪多糖组的1.490倍,差异极显著(P<0.01),复方苦芩组、黄芪多糖组BAX基因转录量分别为空白组的1.116倍、0.966倍,差异不显著(P>0.05);与空白组相比,模型组Bcl-2/BAX mRNA转录比值降低,差异极显著(P<0.01);与模型组相比,复方苦芩组、黄芪多糖组Bcl-2/BAX mRNA转录比值均增加,分别为模型组1.809倍和1.940倍,差异极显著(P<0.01),复方苦芩组Bcl-2/BAX mRNA转录比值分

英文摘要:

[Objective] The objective of this study is to investigate the effect of Kuqin compound on the transcription of Bcl-2/BAX mRNA in porcine kidney cells (PK-15) infected with transmissible gastroenteritis virus (TGEV) . [Method] To study the drug (Kuqin compound) test in vitro by TGEV infected PK-15 cell model, the test was divided into Kuqin compound group, blank control group, model group, and astragalus polysaccharide group, respectively. The morphology change was observed in PK-15 cells by the Swiss staining method and the transcription of Bcl-2 and BAX mRNA was tested by RT-PCR method after determination of tissue culture infective dose (TCID50) of TGEV and the maximum of no toxic concentration of Kuqin compound and astragalus polysaccharide on PK-15 cells. [Result]The following results were obtained from the observation. Compared with blank control group, the PK-15 cells in model group had obvious cytopathic effects (CPE) including elongated cell shape, intercellular drawing into mesh, and lighter staining in nucleus. The nucleus enrichment, pyknosis and fragmentation were also observed. On the other hand, compared with the model group, CPE in Kuqin compound and astragalus polysaccharide group was improved and the Kuqin compound had better effect with most nucleus structure clear, uniform staining in chromatin and fewer nucleuses became larger or fragmentized. The transcription amounts were also considered as an effect of compound Kuqin on infected PK-15 cells. Compared with blank, the transcription amount of Bcl-2 mRNA in model group was less (P〈0.01). However, the transcription amount of Bcl-2 mRNA in compound Kuqin group was increased by 1.403 times compared with the model group (P〈0.01), by 1.078 times with the Astragalus polysaccharide group (P〉0.05). For the BAX mRNA transcription, the model group had an obvious increase (increased by 1.440 times, 1.290 times and 1.490 times compared with the blank group, compound Kuqing group and the astragalus polysaccharide grou

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期刊信息
  • 《中国农业科学》
  • 中国科技核心期刊
  • 主管单位:中华人民共和国农业部
  • 主办单位:中国农业科学院 中国农学会
  • 主编:万建民
  • 地址:北京中关村南大街12号中国农业科学院图书馆楼4101-4103室
  • 邮编:100081
  • 邮箱:zgnykx@caas.cn
  • 电话:010-82109808 82106279
  • 国际标准刊号:ISSN:0578-1752
  • 国内统一刊号:ISSN:11-1328/S
  • 邮发代号:2-138
  • 获奖情况:
  • 中国期刊方阵“双高”期刊,第三届中国出版政府奖提名奖
  • 国内外数据库收录:
  • 美国化学文摘(网络版),英国农业与生物科学研究中心文摘,波兰哥白尼索引,英国动物学记录,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),英国食品科技文摘,中国北大核心期刊(2000版)
  • 被引量:85620