Antiapoptotic and Antiautophagic Effects of Glial Cell Line-Derived Neurotrophic Factor and Hepatocyte Growth Factor After Transient Middle Cerebral Artery Occlusion in Rats
Antiapoptotic and Antiautophagic Effects of Glial Cell Line-Derived Neurotrophic Factor and Hepatocyte Growth Factor After Transient Middle Cerebral Artery Occlusion in Rats
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DOI：
10.1002/jnr.22373
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发表时间：
2010-08-01
影响因子：
4.2
通讯作者：
Abe, Koji
中科院分区：
文献类型：
--
作者：
Shang, Jingwei;Deguchi, Kentaro;Abe, Koji
胶质细胞源性神经营养因子(GDNF)和肝细胞生长因子(HGF)是一类强神经营养因子，具有抗细胞凋亡的作用。然而，GDNF和HGF通过抗自噬作用改善缺血性脑损伤的神经保护作用尚未得到检验。因此，我们研究了GDNF和HGF对大鼠大脑中动脉短暂性闭塞(TMCAO)后脑梗塞面积和抗细胞凋亡、抗自噬作用的影响。用HE染色计算TMCAO后24小时的脑梗塞面积，判断缺血性脑损伤的程度。用末端脱氧核苷酸转移酶介导的dUTP-生物素原位缺口末端标记法(TUNEL)评价其抗细胞凋亡作用。用Western印迹分析微管相关蛋白1轻链3(LC3)，免疫荧光分析LC3和磷酸化Nitor/Ser(2448)(p-mTOR)来评价其抗自噬作用。GDNF和HGF可显著缩小脑缺血后的脑梗塞面积。TMCAO后Lc3-I和Lc3-II(相对于p-微管蛋白)的含量显著增加，而GDNF和HGF显著降低。GDNF和HGF可显著增加p-mTOR阳性细胞数。GDNF和HGF可显著减少TUNEL-、LC3-和LC3/TUNEL双阳性细胞数。Lc3/TUNEL双阳性细胞约占Lc3+TUNEL阳性细胞的34.3%。提示GDNF和HGF的保护作用不仅与抗细胞凋亡有关，而且与抗自噬作用密切相关，可能是同一细胞同时发生两种类型的细胞死亡，GDNF和HGF能够改善这两种途径。(C)2010年Wiley-Liss公司
Glial cell line-derived neurotrophic factor (GDNF) and hepatocyte growth factor (HGF) are strong neurotrophic factors, which function as antiapoptotic factors. However, the neuroprotective effect of GDNF and HGF in ameliorating ischemic brain injury via an antiautophagic effect has not been examined. Therefore, we investigated GDNF and HGF for changes of infarct size and antiapoptotic and antiautophagic effects after transient middle cerebral artery occlusion (tMCAO) in rats. For the estimation of ischemic brain injury, the infarct size was calculated at 24 hr after tMCAO by HE staining. Terminal deoxynucleotidyl transferase-mediated dUTP-biotin in situ nick end labeling (TUNEL) was performed for evaluating the antiapoptotic effect. Western blot analysis of microtubule-associated protein 1 light chain 3 (LC3) and immunofluorescence analysis of LC3 and phosphorylated niTOR/Ser(2448) (p-mTOR) were performed for evaluating the antiautophagic effect. GDNF and HGF significantly reduced infarct size after cerebral ischemia. The amounts of LC3-I plus LC3-II (relative to p-tubulin) were significantly increased after tMCAO, and GDNF and HGF significantly decreased them. GDNF and HGF significantly increased p-mTOR-positive cells. GDNF and HGF significantly decreased the numbers of TUNEL-, LC3-, and LC3/TUNEL double-positive cells. LC3/TUNEL double-positive cells accounted for about 34.3% of LC3 plus TUNEL-positive cells. This study suggests that the protective effects of GDNF and HGF were greatly associated with not only the antiapoptotic but also the antiautophagic effects; maybe two types of cell death can occur in the same cell at the same time, and GDNF and HGF are capable of ameliorating these two pathways. (C) 2010 Wiley-Liss, Inc.