Pan Qin, Zhang Zhong-Bing, Zhang Xin, Shi Jian, Chen Yue-Xiang, Han Ze-Guang, Xie Wei-Fen
Department of Gastroenterology, Changzheng Hospital, Second Military Medical University, 415 Fengyang Road, Shanghai 200003, PRC.
Dig Dis Sci. 2007 Oct;52(10):2591-600. doi: 10.1007/s10620-006-9676-1. Epub 2007 Mar 16.
Our aim was to gain insight into the gene expression profile during hepatic fibrosis autoreversal. Spontaneous recovery from hepatic fibrosis was created in SD rats by CCl(4) exposure for 8 weeks and then withdrawal for 6 weeks. Then differentially expressed genes during regression of fibrosis were analyzed using cDNA microarray. Results obtained were further subjected to hierarchical clustering and validated by semiquantitative RT-PCR. Expression of Mapk1 and Rps6ka1, which are critical members of the mitogen-activated protein kinase (MAPK) signaling pathway, was also investigated by Northern blot and immunohistochemistry. Microarray hybridization identified 254 genes differentially expressed throughout resolution of fibrosis. Being verified by RT-PCR, up- or down-regulated genes were classified into various groups according to clustering and function: (1) metabolic enzymes, (2) facilitated diffusion proteins/transporters/symporters, (3) gastrointestinal hormones/receptors, (4) lipoproteins/fatty acid binding proteins, (5) transcription factors/nuclear factors, and (6) the MAPK signaling pathway. The mRNA level of Mapk1 increased greatly as hepatic fibrosis reversed. Meanwhile Mapk1 and Rps6ka1 were proven to be expressed in hepatocytes and absent from mesenchymal cells. Six groups of genes exhibit a close relation to the recovery of CCl(4)-induced hepatic fibrosis. The MAPK signaling-dependent pathway, representing one of the gene groups, may contribute to the reversal of hepatic fibrosis.
我们的目的是深入了解肝纤维化自动逆转过程中的基因表达谱。通过对SD大鼠进行8周的四氯化碳（CCl₄）暴露，然后停药6周，诱导肝纤维化自发恢复。然后使用cDNA微阵列分析纤维化消退过程中差异表达的基因。对获得的结果进行层次聚类，并通过半定量逆转录聚合酶链反应（RT-PCR）进行验证。还通过Northern印迹和免疫组织化学研究了丝裂原活化蛋白激酶（MAPK）信号通路的关键成员Mapk1和Rps6ka1的表达。微阵列杂交鉴定出254个在纤维化消退过程中差异表达的基因。经RT-PCR验证后，上调或下调的基因根据聚类和功能分为不同组：（1）代谢酶，（2）易化扩散蛋白/转运蛋白/同向转运体，（3）胃肠激素/受体，（4）脂蛋白/脂肪酸结合蛋白，（5）转录因子/核因子，以及（6）MAPK信号通路。随着肝纤维化的逆转，Mapk1的mRNA水平大幅增加。同时，Mapk1和Rps6ka1被证明在肝细胞中表达，而在间充质细胞中不表达。六组基因与CCl₄诱导的肝纤维化的恢复密切相关。代表基因组之一的MAPK信号依赖性途径可能有助于肝纤维化的逆转。
