单一延长应激诱导创伤后应激障碍大鼠海马内质网依赖性细胞凋亡。
Single-prolonged stress induces endoplasmic reticulum-dependent apoptosis in the hippocampus in a rat model of post-traumatic stress disorder.
机构信息
PTSD Laboratory, Department of Histology and Embryology, Institute of Pathology and Pathophysiology, China Medical University, Shenyang, China.
出版信息
PLoS One. 2013 Jul 19;8(7):e69340. doi: 10.1371/journal.pone.0069340. Print 2013.
BACKGROUND
Our previous research indicated that apoptosis induced atrophy in the hippocampus of post-traumatic stress disorder (PTSD) rats. Endoplasmic reticulum (ER) stress-induced apoptosis has been implicated in the development of several disorder diseases. The aim of this study was to investigate whether endoplasmic reticulum-related pathway is involved in single-prolonged stress (SPS) induces apoptosis in the hippocampus of PTSD rats by examining the expression levels of three important indicators in the ER-related apoptotic pathway: Glucose-regulated protein (GRP) 78, caspase-12 and Ca(2+)/CaM/CaMkinaseIIα (CaMkIIα).
METHODS
Wistar rats were sacrificed at 1, 4 and 7 days after SPS. SPS is a reliable animal model of PTSD. The apoptotic cells in the hippocampus were assessed by TUNEL method and transmission electron microscopy (TEM). Free intracellular Ca(2+) concentration was measured. GRP78 expression was examined by immunohistochemistry, western blotting and RT-PCR. mRNA of caspase-12 and CaM/CaMkIIα were determined by RT-PCR.
RESULTS
Our results showed that apoptotic cells were increased in the SPS rats. TEM analysis revealed characteristic morphological changes of apoptosis in these cells. We observed that GRP78 was significantly up-regulated during early PTSD, and then recovered at 7 days after SPS. By RT-PCR, we observed that the change in caspase-12 expression level was similar to that in GRP78. Moreover, the free intracellular Ca(2+) concentration was significantly higher at 1 day after SPS and decreased in 7 days. CaM expression increased significantly, while CaMKIIα expression decreased significantly in the hippocampus at 1 day after SPS.
CONCLUSION
SPS induced change in the expression levels of GRP78, caspase-12 and Ca(2+)/CaM/CaMkIIα in the hippocampus of PTSD rats indicated that the endoplasmic reticulum pathway may be involved in PTSD-induced apoptosis.
背景
我们之前的研究表明，创伤后应激障碍（PTSD）大鼠海马区的凋亡诱导萎缩。内质网（ER）应激诱导的细胞凋亡与多种疾病的发生发展有关。本研究旨在通过检测内质网相关凋亡途径中的三个重要指标的表达水平，探讨内质网相关途径是否参与单延长应激（SPS）诱导 PTSD 大鼠海马区细胞凋亡：葡萄糖调节蛋白（GRP）78、半胱天冬酶-12 和 Ca（2+）/CaM/CaMkinaseIIα（CaMkIIα）。
方法
SPS 后 1、4、7 天处死 Wistar 大鼠。SPS 是 PTSD 的可靠动物模型。通过 TUNEL 法和透射电镜（TEM）评估海马区凋亡细胞。测量细胞内游离 Ca（2+）浓度。通过免疫组化、Western blot 和 RT-PCR 检测 GRP78 表达。通过 RT-PCR 测定 caspase-12 和 CaM/CaMkIIα 的 mRNA。
结果
我们的结果表明，SPS 大鼠海马区凋亡细胞增多。TEM 分析显示这些细胞中存在凋亡的特征形态变化。我们观察到 GRP78 在早期 PTSD 中显著上调，然后在 SPS 后 7 天恢复。通过 RT-PCR，我们观察到 caspase-12 表达水平的变化与 GRP78 的变化相似。此外，SPS 后 1 天细胞内游离 Ca（2+）浓度显著升高，7 天降低。SPS 后 1 天，海马 CaM 表达显著增加，CaMKIIα 表达显著降低。
结论
SPS 诱导 PTSD 大鼠海马区 GRP78、caspase-12 和 Ca（2+）/CaM/CaMkIIα 表达水平的变化表明内质网途径可能参与 PTSD 诱导的细胞凋亡。
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