GZMA、GZMB、TSP-1、TLR和IL-15条细胞凋亡通路基因在肝再生中的表达变化

Expression changes of GZMA,GZMB,TSP-1,TLR and IL-1 five cell apoptosis pathways related genes in liver regeneration

目的探讨颗粒酶A(GZMA)、颗粒酶B(GZMB)、血小板反应蛋白-1(TSP-1)、Toll样受体(TLR)和白细胞介素-1(IL-1)5条细胞凋亡通路基因在肝再生(LR)过程中的表达变化。方法大鼠随机分为33组,每组6只,用Rat Genome 230 2.0芯片检测大鼠部分肝切除(PH)后不同恢复时间点5条细胞凋亡通路基因的表达情况,采用Real-time PCR和Western blotting技术对芯片结果进行验证,并用生物信息学方法对凋亡通路基因在肝再生中的表达变化进行分析。结果 Real-time PCR和Western blotting均与Rat Genome 230 2.0芯片的检测结果趋势一致;GZMA、GZMB、TSP-1、TLR和IL-1 5条细胞凋亡通路中9、8、24、31和34个基因与肝再生相关。它们主要在肝再生启动阶段起始表达,在细胞增殖阶段表达的基因数最多。表达的相似性分为均上调、上调占优势、均下调、下调占优势、上调和下调相近等5类,大多数基因表达加强,少数基因表达降低。它们表达的时间相关性分为13组。基因协同作用模型(Et)分析表明,GZMA介导的细胞凋亡通路在肝再生后期促进细胞凋亡;TLR介导的细胞凋亡通路几乎均在整个肝再生中促进细胞凋亡;GZMB、TSP-1和IL-1介导的细胞凋亡通路可能在肝再生中不发挥细胞凋亡作用。结论 GZMA和TLR两条通路调控肝再生中的细胞凋亡。

Objective To investigate the expression changes of five cell apoptosis pathway genes,granzyme A（ GZMA）,granzyme B（ GZMB）,thrombospondin-1（ TSP-1）,Toll-like receptors（ TLR） and interleukin-1（ IL-1）,in the process of regeneration liver（ LR）. Methods Rats were randomly divided into 33 groups with 6 rats in each group.Genome Rat 230 2. 0 was used to detect the expression of genes in four cell apoptosis pathways in rats after partial hepatectomy（ PH）. Real-time PCR and Western blotting technology were used to verify the results of the chip,and the expression changes of apoptosis pathway genes in liver regeneration were analyzed by bioinformatics methods. Results Both Real-time PCR and Western blotting were in agreement with the test results of Genome Rat 230 2. 0 chip; 9,8,24,31 and 34 genes were associated with liver regeneration in the four cell apoptosis pathways. They were mainly expressed at the start stage of liver regeneration. The maximum number of expressed genes were in cell proliferation phase. The similarity of their expression was totally up regulation,up regulation advantage,totally down regulation,down regulation advantage,up regulation equal to down regulation of 5 categories. The expression of most genes in liver regeneration was enhanced,and the expression of few gene was decreased. Time correlation was divided into 13 groups. Analysis of the role of apoptosis pathway related genes in liver regeneration by using the gene synergy model（ Et）,GZMA pathway promotedapoptosis in the late stage of liver regeneration; TLR pathway promotes cell apoptosis almost in the whole liver regeneration;GZMB,TSP-1 and IL-1 pathways did not play the role of apoptosis in liver regeneration. Conclusion Two apoptosis pathways,GZMA and TSP-1,regulate apoptosis in liver regeneration.