Zhao Ya-Ning, Wang Hong-Yang, Li Jian-Min, Chen Bao-Yuan, Xia Guo, Zhang Pan-Pan, Ge Yan-Lei
Department of Rehabilitation, Affiliated Hospital of Hebei United University, Tangshan, Hebei 063000, P.R. China.
Department of Respiratory Medicine, Affiliated Hospital of Hebei United University, Tangshan, Hebei 063000, P.R. China.
Mol Med Rep. 2016 Jan;13(1):137-45. doi: 10.3892/mmr.2015.4505. Epub 2015 Nov 5.
Obstructive sleep apnea syndrome (OSAS), characterized by intermittent hypoxia/re‑oxygenation, may impair the cerebral system. Although mitogen‑activated protein kinase (MAPK) signaling was observed to have a key role in hypoxia‑induced brain injury, the intracellular events and their underlying mechanisms for intermittent hypoxia/re‑oxygenation-associated damage to hippocamal MAPKs, including extracellular signal‑regulated kinase (ERK)1/2, P38MAPK and c‑Jun N‑terminal kinase (JNK) remain to be elucidated and require further investigation. A total of five rats in each sub‑group were exposed to intermittent hypoxia or continued hypoxia for 2, 4, 6 or 8 weeks. Histological, immunohistochemical and biological analyses were performed to assess nerve cell injury in the hippocampus. Surviving CA1 pyramidal cells were identified by hematoxylin and eosin staining. The levels of phosphorylated ERK1/2, P38MAPK and JNK were detected by western blotting. B‑cell lymphoma 2 (Bcl‑2) and Bcl‑2‑associated X protein (Bax) in neural cells were examined by immunohistochemistry. The malondialdehyde (MDA) contents and superoxide dismutase (SOD) activities were measured by thiobarbituric acid and xanthine oxidation methods, respectively. Under continued hypoxia, the levels of phospho‑ERK1/2 peaked at the fourth week and then declined, whereas phospho‑P38MAPK and JNK were detected only in the late stages. By contrast, under intermittent hypoxia, ERK1/2, P38MAPK and JNK were activated at all time-points assessed (2, 4, 6 and 8 weeks). The levels of phospho‑ERK1/2, P38MAPK and JNK were all higher in the intermittent hypoxia groups than those in the corresponding continued hypoxia groups. Bcl‑2 was mainly increased and reached the highest level at six weeks in the continued hypoxia group. Of note, Bcl‑2 rapidly increased to the peak level at four weeks, followed by a decrease to the lowest level at the eighth week in the intermittent hypoxia group. Bax was generally increased at the late stages under continued hypoxia, but increased at all time-points under the intermittent hypoxia conditions. The two types of hypoxia induced an increase in the MDA content, but a decrease in SOD activity. Marked changes in these two parameters coupled with markedly reduced surviving cells in the hippocampus in a time‑dependent manner were observed in the intermittent hypoxia group in comparison with the continued hypoxia group. OSAS‑induced intermittent hypoxia markedly activated the MAPK signaling pathways, which were triggered by oxidative stress, leading to abnormal expression of downstream Bcl‑2 and Bax, and a severe loss of neural cells in the hippocampus.
阻塞性睡眠呼吸暂停综合征(OSAS)以间歇性缺氧/复氧为特征,可能会损害脑系统。尽管有研究观察到丝裂原活化蛋白激酶(MAPK)信号通路在缺氧诱导的脑损伤中起关键作用,但间歇性缺氧/复氧相关的海马MAPK损伤(包括细胞外信号调节激酶(ERK)1/2、P38MAPK和c-Jun氨基末端激酶(JNK))的细胞内事件及其潜在机制仍有待阐明,需要进一步研究。每个亚组共有5只大鼠暴露于间歇性缺氧或持续性缺氧环境中2、4、6或8周。进行组织学、免疫组织化学和生物学分析以评估海马中的神经细胞损伤。通过苏木精和伊红染色鉴定存活的CA1锥体细胞。通过蛋白质印迹法检测磷酸化ERK1/2、P38MAPK和JNK的水平。通过免疫组织化学检测神经细胞中的B细胞淋巴瘤2(Bcl-2)和Bcl-2相关X蛋白(Bax)。分别采用硫代巴比妥酸法和黄嘌呤氧化法测定丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性。在持续性缺氧条件下,磷酸化ERK1/2水平在第4周达到峰值,然后下降,而磷酸化P38MAPK和JNK仅在后期被检测到。相比之下,在间歇性缺氧条件下,ERK1/2、P38MAPK和JNK在所有评估时间点(2、4、6和8周)均被激活。间歇性缺氧组中磷酸化ERK1/2、P38MAPK和JNK的水平均高于相应的持续性缺氧组。在持续性缺氧组中,Bcl-2主要升高,并在第6周达到最高水平。值得注意的是,在间歇性缺氧组中,Bcl-2在第4周迅速升高至峰值水平,随后在第8周降至最低水平。在持续性缺氧后期,Bax总体上升高,但在间歇性缺氧条件下,Bax在所有时间点均升高。两种类型的缺氧均导致MDA含量增加,但SOD活性降低。与持续性缺氧组相比,间歇性缺氧组中这两个参数的显著变化以及海马中存活细胞数量随时间的显著减少均被观察到。OSAS诱导的间歇性缺氧显著激活了MAPK信号通路,该通路由氧化应激触发,导致下游Bcl-2和Bax异常表达以及海马中神经细胞的严重损失。