【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转录比值分别为空白组的0.749倍（P＜0.01）、黄芪多糖组的0.933倍（P＞0.05）。【结论】复方苦芩可通过上调PK-15细胞Bcl-2 mRNA转录，下调BAX mRNA的转录，从而抑制TGEV引起的PK-15细胞凋亡。%[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 (P0.05). 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 group) while no obvious changes among the other three groups (compound Kuqin was 1.116 larger than blank and astragalus polysaccharide was 0.966 of the blank). Moreover, the transcription ratio of Bcl-2/BAX in model group had a huge decrease compared with the blank group (P<0.01). This ratio in compound Kuqin and astragalus polysaccharide groups was much larger compared with the model group (P<0.01) with the number of 1.809 and 1.904 times, respectively;this number was 0.749 (P<0.01) and 0.933 times (P<0.05) of blank, respectively.[Conclusion]Kunqin compound could inhibit apoptosis in PK-15 cells infected by TGEV by up-regulating transcription of Bcl-2 mRNA and down-regulating transcription of BAX mRNA.
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