解剖学报

解剖学报 ›› 2015, Vol. 46 ›› Issue (1): 25-31.doi: 10.16098/j.issn.0529-1356.2015.01.005

• 神经生物学 • 上一篇    下一篇

13-甲基十四烷酸对氧反常诱导大鼠胚脑皮质神经元凋亡和形态学损伤的保护作用

余涓1,胡桂芳1,翁绳美2,巢苹1   

  1. 1.福建医科大学基础医学院生理学与病理生理学系; 2.药学院药理学系， 福州 350004
  • 收稿日期:2014-09-26 修回日期:2014-11-07 出版日期:2015-02-06 发布日期:2015-02-06
  • 通讯作者: 余涓 E-mail:tune9@163.com
  • 基金资助:

    福建省科技厅计划项目资助基金;福建医科大学教授基金资助项目

Protective effects of 13-methyl tetradecanoic acid on the apoptosis of rat embryonic cortical neurons and the morphological damage induced by oxygen paradox

YU Juan 1* HU Gui-fang1 WENG Sheng-mei2 CHAO Ping1   

  1. 1. Department of Physiology and Pathophysiology of Basic Medical College;2. Department of Pharmacology of Pharmacy College, Fujian Medical University, Fuzhou 350004, China
  • Received:2014-09-26 Revised:2014-11-07 Online:2015-02-06 Published:2015-02-06
  • Contact: YU Juan E-mail:tune9@163.com

摘要：

目的 探讨13-甲基十四烷酸(13-MTD)对氧反常诱导大鼠胚脑皮质神经元凋亡和形态学损伤的保护作用。 方法 原代培养大鼠胚脑皮质神经元，并以神经元特异性烯醇化酶（NSE）免疫荧光法鉴定。将神经细胞随机分为正常对照组、模型组和不同剂量13-MTD组，每组设6个复孔。氧糖剥夺3h/再复氧糖24h(OGD3h/R24h)方法制备神经元氧反常模型，于再复氧糖即刻分别给予13-MTD 5、10、20、40mg/L干预。倒置相差显微镜下观察神经细胞形态改变；磺酰罗丹明B（SRB）法测定神经细胞存活率； 吖啶橙/溴乙啶（AO/EB）染色法观察神经细胞凋亡；透射电子显微镜下观察神经元超微结构的改变。结果 与正常组比较，模型组神经元呈现病理改变，神经细胞存活率显著下降(P<0.01)，细胞凋亡显著增多(P <0.01)，神经元超微结构损伤明显，可见核内染色质边集或凝聚成块状，胞质内细胞器明显减少甚至消失，线粒体肿胀、嵴断裂甚至消失等；与模型组比较，不同剂量的13-MTD可有效改善上述变化(P<0.05, P<0.01)，使神经元形态及其超微结构损伤明显恢复，且存在剂量依赖关系，以13-MTD 20mg/L 改善更显著(P<0.01)。 结论 13-MTD对氧反常诱导的大鼠胚脑皮质神经元损伤具有明显保护作用，其可能通过改善神经细胞形态和线粒体超微结构损伤，减少神经元凋亡，提高细胞存活率。

关键词: 13-甲基十四烷酸, 氧反常, 超微结构损伤, 皮质神经元, 免疫荧光, 大鼠

Abstract:

Objective To investigate the protective effect of 13-methyltetradecanoic acid(13-MTD) on the apoptosis and morphological damage of primary cultured rat embryonic cortical neurons induced by the oxygen paradox. Methods Primary culturing rat embryonic cortical neurons were identified by NSE immunefluorescence. They were randomly divided into normal control group, model group and the different doses of 13-MTD groups. Each group contained 6 subholes. The model of oxygen paradox of oxygenglucose deprivation for 3 hours/reperfusion for 24 hours (OGD 3h/R 24h) was made. 13MTD 5mg/L, 10mg/L, 20mg/L, 40mg/L were administered immediately after reperfusion. An inverted phase contrast microscope was used to observe the danamic morphology of the cells. Sulforhodamine B (SRB) assay was applied to detect the cellular survival rate. Acridine orange/ethidium bromide(AO/EB) double staining was adopted to observe the apoptotic morphology of neurons. The transmission electron microscope was adopted to observe the changes of ultrastructure in primary cultured rat embryonal cerebral cortical neurons.Results Compared with the normal control group, the model group showed that the pathological damage was apparent, the nerve cell survival rate decreased significantly(P<0.01), the neuron apoptosis rate was increased dramatically (P<0.01), and the ultrastructure of neurons was injured seriously. The nuclear chromatin was focused on the surrounding or condensed into squares, the cytoplasmic organelles reduced or even disappeared significantly, mitochondria was swelling and its crests was break and even disappeared. Compared with the model group, different doses of 13-MTD improved the above changes significantly(P<0.05, P<0.01), the neuronal morphology and its ultrastructural damage were restored with a dose-dependent manner, and 13-MTD 20μg/ml was improved more significant(P<0.01). Conclusion 13-MTD has obvious protective effects on the rat embryonic cortical neuron damage induced by oxygen paradox, which may act through the improvement of nerve cell morphology and mitochondrial ultrastructure damage, thereby decreasing neuronal apoptosis and increasing the survival rate of neurons.

Key words: 13-Methyl tetradecanoic acid, Oxygen paradox, Ultrastructure injury, Cortical neuron, Immunofluorescence, Rat