Enhanced anti-apoptosis and gut epithelium protection function of acidic fibroblast growth factor after cancelling of its mitogenic activity
Enhanced anti-apoptosis and gut epithelium protection function of acidic fibroblast growth factor after cancelling of its mitogenic activity
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DOI：
10.3748/wjg.v10.i24.3590
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发表时间：
2004-12-15
影响因子：
4.3
通讯作者：
Sheng, Zhi-Yong
中科院分区：
文献类型：
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作者：
Fu, Xiao-Bing;Li, Xiao-Kun;Sheng, Zhi-Yong
目的：成纤维细胞生长因子（FGF）的促有丝分裂和非促有丝分裂活性与一系列生物学功能相关，从细胞增殖和分化到许多疾病的发生。近年来研究表明酸性成纤维细胞生长因子（aFGF）具有较强的抗凋亡作用，可能对肠缺血再灌注损伤具有潜在的治疗作用。然而，这种功能是否取决于其促有丝分裂或非促有丝分裂活性仍不清楚。方法：扩增aFGF 28 -154基因片段，亚克隆入pET-3c载体，在BL 21（DE 3）细胞中表达，经M2琼脂糖亲和柱纯化，获得aFGF 28 -154基因突变体。该突变体aFGF 28 -154保持其非促有丝分裂活性并丧失其促有丝分裂活性。采用地塞米松（dexamethasone，DEX）诱导的小鼠胸腺细胞凋亡模型，研究aFGF 28 -154的抗凋亡作用。此外，进行体内研究以进一步证实aFGF 28 -154是否可以显著减少大鼠肠缺血-再灌注损伤后肠上皮细胞的凋亡。结果：在地塞米松（dexamethasone，DEX）诱导的小鼠胸腺细胞凋亡模型中，aFGF 28 -154的抗凋亡作用较野生型aFGF明显增强。体内研究进一步证实，aFGF 28 -154显著减少大鼠肠缺血-再灌注损伤后肠上皮细胞的凋亡。aFGF 28 -154的抗凋亡作用机制不依赖于其促有丝分裂活性，主要与其非促有丝分裂活性有关，包括保护细胞内钙离子平衡、维持ERK 1/2激活和细胞周期平衡。这些发现强调了aFGF的非促有丝分裂作用的重要性，并暗示其在预防由缺血-再灌注损伤引发的组织和内部器官中的细胞凋亡和其它损伤中的治疗用途。
AIM: Mitogenic and non-mitogenic activities of fibroblast growth factor (FGF) are coupled to a range of biological functions, from cell proliferation and differentiation to the onset of many diseases. Recent reports have shown that acidic fibroblast growth factor (aFGF) has a powerful antiapoptosis function, which may have potentially therapeutical effect on gut ischemia and reperfusion injuries. However, whether this function depends on its mitogenic or non-mitogenic activity remains unclear. In this study, we identified the source of its anti-apoptosis function with a mutant, aFGF28-154 and observed its effect on reducing gut ischemia and reperfusion injury.METHODS: aFGF28-154 was generated by amplification of appropriate DNA fragments followed by subcloning the products into pET-3c vectors, then they were expressed in BL21 (DE3) cells and purified on an M2 agarose affinity column. This mutant aFGF28-154 maintained its non-mitogenic activity and lost its mitogenic activity. With a dexamethasone (DEX)-induced mouse thymocyte apoptosis model in vitro and in vivo, we studied the anti-apoptotic function of aFGF28-154. Also, in vivo study was performed to further confirm whether aFGF28-154 could significantly reduce apoptosis in gut epithelium after gut ischemia-reperfusion injury in rats. Based on these studies, the possible signal transduction pathways involved were studied.RESULTS: With a dexamethasone (DEX)-induced mouse thymocyte apoptosis model in vitro and in vivo, we found that the anti-apoptotic function of aFGF28-154 was significantly enhanced when compared with the wild type aFGF. In vivo study further confirmed that aFGF28-154 significantly reduced apoptosis in gut epithelium after gut ischemia-reperfusion injury in rats. The mechanisms of anti-apoptosis function of aFGF28-154 did not depend on its mitogenic activity and were mainly associated with its non-mitogenic activities, including the intracellular calcium ion balance protection, ERK1/2 activation sustaining and cell cycle balance.CONCLUSION: These findings emphasize the importance of non-mitogenic effects of aFGF, and have implications for its therapeutic use in preventing apoptosis and other injuries in tissues and internal organs triggered by ischemia-reperfusion injury.