CAJ | 학술논문

背景:神经元本身因缺血引起损伤以及缺血后再灌注导致神经元损伤是缺血性脑损伤的两大原因,二者均伴有神经元胞质内钙离子浓度增高,这种变化的重要意义备受研究者的关注,可能是各种损伤因素作用的结果,也可能是造成脑缺血损伤的各种因素最后作用的共同通路,即各种因素最后是通过增高的钙离子来产生神经毒性.目的:探讨缺血性神经元的损伤与细胞内钙离子的关系及氟桂利嗪的治疗意义.设计:完全随机设计,对照实验研究.地点和材料:解放军第四军医大学西京医院神经内科实验室和第四军医大学电镜中心.实验材料为孕15 d昆明种二级孕鼠由本校实验动物中心提供,清洁级.干预:将培养的神经元随机分为缺血组,氟桂利嗪预处理组以及对照组,利用Ca2+指示剂Flu-3/AM作为细胞内钙离子的荧光探针负载培养的神经元,共聚焦技术检测细胞内荧光强度的变化,利用四唑蓝(四唑蓝)比色试验检测缺血性神经元的损伤程度.缺血组神经元给予类缺血处理10 min,氟桂利嗪组2 min预处理后,类缺血处理10 min.对照组给予含糖的人工脑脊液,混合气体为50 mL/L CO2的氧气,10 min后进行实验.干预者为作者本人.主要观察指标:观察各组神经元经相应处理后,神经元胞浆内钙离子浓度的变化,细胞活性变化.结果:给予神经元类缺血处理10 min后,缺血组神经元胞质内钙离子浓度(平均荧光强度变化为17.525±3.167)和细胞损伤程度(吸光度为0.175±0.021)明显高于氟桂利嗪预处理组(P＜0.01).结论:氟桂利嗪可明显抑制神经元胞质内钙离子的升高,减轻由此引发的缺血性神经元的损伤程度.
배경:신경원본신인결혈인기손상이급결혈후재관주도치신경원손상시결혈성뇌손상적량대원인,이자균반유신경원포질내개리자농도증고,저충변화적중요의의비수연구자적관주,가능시각충손상인소작용적결과,야가능시조성뇌결혈손상적각충인소최후작용적공동통로,즉각충인소최후시통과증고적개리자래산생신경독성.목적:탐토결혈성신경원적손상여세포내개리자적관계급불계리진적치료의의.설계:완전수궤설계,대조실험연구.지점화재료:해방군제사군의대학서경의원신경내과실험실화제사군의대학전경중심.실험재료위잉15 d곤명충이급잉서유본교실험동물중심제공,청길급.간예:장배양적신경원수궤분위결혈조,불계리진예처리조이급대조조,이용Ca2+지시제Flu-3/AM작위세포내개리자적형광탐침부재배양적신경원,공취초기술검측세포내형광강도적변화,이용사서람(사서람)비색시험검측결혈성신경원적손상정도.결혈조신경원급여류결혈처리10 min,불계리진조2 min예처리후,류결혈처리10 min.대조조급여함당적인공뇌척액,혼합기체위50 mL/L CO2적양기,10 min후진행실험.간예자위작자본인.주요관찰지표:관찰각조신경원경상응처리후,신경원포장내개리자농도적변화,세포활성변화.결과:급여신경원류결혈처리10 min후,결혈조신경원포질내개리자농도(평균형광강도변화위17.525±3.167)화세포손상정도(흡광도위0.175±0.021)명현고우불계리진예처리조(P＜0.01).결론:불계리진가명현억제신경원포질내개리자적승고,감경유차인발적결혈성신경원적손상정도.
BACKGROUND: Ischemia and ischemia-reperfusion, two main causes for ischemic cerebral impairment, contribute to neuronal injury and are always accompanied by intraneuronal calcium elevation. Thereby significance of intracellular calcium change gains more attention from researchers. Either as the result of multiple nosogenesis or as the last common pathway of cerebral ischemic impairment, elevated intracellular calcium is believed to exert neurotoxic function.OBJECTIVE: To study the changes of intracellular free calcium and cell activity of ischemic neurons, so as to investigate the relationship between ischemic neuronal impairment and intracellular calcium, as well as the therapeutic significance of flunarizin.DESIGN: A completely randomized control study.SETTING and MATERIALS: Neurological Laboratory and Electroscopic Center of Xijing Hospital, Fourth Military Medical University of Chinese PLA. Kunming 15-day pregnant rats, grade two and cleanness grade, were provided by the Experimental Animal Center.INTERVENTION: In vitro cultured neurons were randomly divided into inschemic group, flunarizn treatment group and control group. Flu-3/AM of Ca2 + indicator used as intracellular fluorescent probe was loaded to cultured neurons, and confocal laser scanning technique was adopted to detect the change of fluorescent intensity. Meanwhile, impairment of ischemic neurons was graded by colorimetry. Neurons of ischemic group received ischemic-like treatment for 10 minutes and additional 2-minute pretreatment of flunarizin in flunarizin group. Neurons in control group were cultured in artificial CSF composed of glucose and mixed oxygen of 50 mL/L CO2 for 10 minutes. The authors completed all these interventions.MAIN OUTCOME MEASURES: Changes in intracellular Ca2 + concentration and neuronal activity after corresponding treatments.RESULTS: Significantly higher intracellular Ca2 + (mean fluorescent intensity = 17.525 ±3. 167) and more severe neuronal impairment(Absorbency =0. 175±0.021) were found in ischemic group than in flunarizin pretreatment group( P ＜ 0. 01).CONCLUSION: Ftunarizin can remarkably inhibit neuronal intracellular Ca2 + elevation, consequently attenuating ischemic neuronal impairment.