GZMA、GZMB、TSP-1、TLR和IL-1 5条细胞凋亡通路基因在肝再生中的表达变化

邢雪琨 刘振华 李梦华 徐存拴*

doi:10.16098/j.issn.0529-1356.2016.04.022

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解剖学报 ›› 2016, Vol. 47 ›› Issue (4) : 563-571. DOI: 10.16098/j.issn.0529-1356.2016.04.022

生物工程学

GZMA、GZMB、TSP-1、TLR和IL-1 5条细胞凋亡通路基因在肝再生中的表达变化

邢雪琨¹ 刘振华¹ 李梦华² 徐存拴^3*

作者信息 +

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

XING Xue-kun¹ LIU Zhen-hua¹LI Meng-hua² XU Cun-shuan^3*

Author information +

文章历史 +

摘要

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。

Abstract

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 promoted apoptosis 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.

导出引用

邢雪琨刘振华李梦华徐存拴. GZMA、GZMB、TSP-1、TLR和IL-1 5条细胞凋亡通路基因在肝再生中的表达变化[J]. 解剖学报. 2016, 47(4): 563-571 https://doi.org/10.16098/j.issn.0529-1356.2016.04.022

XING Xue-kun LIU Zhen-hua LI Meng-hua XU Cun-shuan. Expression changes of GZMA, GZMB, TSP-1, TLR and IL-1 five cell apoptosis pathways related genes in liver regeneration[J]. Acta Anatomica Sinica. 2016, 47(4): 563-571 https://doi.org/10.16098/j.issn.0529-1356.2016.04.022

参考文献

［1］Jiang WW, Hao XX, Guo YQ, et al. Caspase signaling pathway regulates hepatocyte apoptosis in rat liver regeneration ［J］. Acta Anatomica Sinica, 2012, 43(3): 359-365. (in Chinese)

蒋文文, 郝晓霞, 郭雅琼, 等. Caspase信号通路调控大鼠再生肝肝细胞的凋亡［J］. 解剖学报, 2012, 43(3): 359-365.

［2］Shen YH, Lu Q, Yang YJ, et al. JNK signaling pathway regulates proliferation and apoptosis of eight liver cell types during rat liver regeneration ［J］. Acta Anatomica Sinica, 2012, 43(2): 189-197. (in Chinese)

申亚慧, 陆琼, 杨彦杰, 等. JNK信号通路调控大鼠再生肝8种细胞的增殖和凋亡［J］. 解剖学报, 2012, 43(2): 189-197.

［3］Lu F, Lamontagne J, Sun A, et al. Role of the inflammatory protein serine protease inhibitor Kazal in preventing cytolytic granule granzyme A-mediated apoptosis ［J］. Immunology, 2011, 134(4): 398-408.

［4］Jacquemin G, Margiotta D, Kasahara A, et al. Granzyme B-induced mitochondrial ROS are required for apoptosis ［J］. Cell Death Differ, 2015, 22(5): 862-874.

［5］El Btaouri H, Morjani H, Greffe Y, et al. Role of JNK/ATF-2 pathway in inhibition of thrombospondin-1 (TSP-1) expression and apoptosis mediated by doxorubicin and camptothecin in FTC-133 cells ［J］. Biochim Biophys Acta, 2011, 1813(5): 695-703.

［6］Panneerselvam P, Ding JL. Beyond TLR signaling-the role of SARM in antiviral immune defense, apoptosis & development ［J］. Int Rev Immunol, 2015, 34(5): 432-444.

［7］Yang B, Elias JE, Bloxham M, et al. Synthetic small interfering RNA down-regulates caspase-3 and affects apoptosis, IL-1 beta, and viability of porcine proximal tubular cells ［J］. J Cell Biochem, 2011, 112(5): 1337-1347.

［8］Xu C, Yang Y, Yang J, et al. Analysis of the role of the integrin signaling pathway in hepatocytes during rat liver regeneration ［J］. Cell Mol Biol Lett, 2012, 17(2): 274-288.

［9］Sakahira H, Enari M, Nagata S. Corrigendum: Cleavage of CAD inhibitor in CAD activation and DNA degradation during apoptosis ［J］. Nature, 2015, 526(7575): 728.

［10］Park JH, Lee SW, Yang SW, et al. Modification of DBC1 by SUMO2/3 is crucial for p53-mediated apoptosis in response to DNA damage ［J］. Nat Commun, 2014, 5: 5483.

［11］Du CP, Tan R, Hou XY. Fyn kinases play a critical role in neuronal apoptosis induced by oxygen and glucose deprivation or amyloid-beta peptide treatment ［J］. CNS Neurosci Ther, 2012, 18(9): 754-761.

［12］Williams ME, Lu X, McKenna WL, et al. UNC5A promotes neuronal apoptosis during spinal cord development independent of netrin-1 ［J］. Nat Neurosci, 2006, 9(8): 996-998.

［13］Andzinski L, Wu CF, Lienenklaus S, et al. Delayed apoptosis of tumor associated neutrophils in the absence of endogenous IFN-beta ［J］. Int J Cancer, 2015, 136(3): 572-583.

［14］Smith MR, Jin F, Joshi I. Milatuzumab and veltuzumab induce apoptosis through JNK signalling in an NF-kappa B dependent human transformed follicular lymphoma cell line ［J］. Br J Haematol, 2014, 165(1): 151-153.

［15］Grenda DS, Murakami M, Ghatak J, et al. Mutations of the ELA2 gene found in patients with severe congenital neutropenia induce the unfolded protein response and cellular apoptosis ［J］. Blood, 2007, 110(13): 4179-4187.

［16］Paul-Samojedny M, Pudelko A, Kowalczyk M, et al. Knockdown of AKT3 and PI3KCA by RNA interference changes the expression of the genes that are related to apoptosis and autophagy in T98G glioblastoma multiforme cells ［J］. Pharmacol Rep, 2015, 67(6): 1115-1123.

［17］Islam TC, Smith CI. The cellular phenotype conditions Btk for cell survival or apoptosis signaling ［J］. Immunol Rev, 2000, 178: 49-63.

［18］Zhou X, Li H, Chai Y, et al. Leptin inhibits the apoptosis of endometrial carcinoma cells through activation of the nuclear factor kB-inducing kinase/lkB kinase Pathway ［J］. Int J Gynecol Cancer, 2015, 25(5): 770-778.