Munc18抗体致痫机制的体外实验研究:诱导海马神经细胞的损伤  

The mechanism of Anti-Munc18 antibody induced epilepsy:antibody induced injury of hippocampal neuron in vitro

作　　者：陈功[1] 江澄川[1] 杨茹[2] 

机构地区：[1]复旦大学附属华山医院神经外科,上海200040 [2]复旦大学神经生物实验室&国家重点实验室,上海200032

出　　处：《中华神经外科疾病研究杂志》2007年第6期491-495,共5页Chinese Journal of Neurosurgical Disease Research

基　　金：国家自然科学基金资助项目(30470536);华山医院基金资助项目(院级-80)

摘　　要：目的前期研究发现将突触前膜胞内蛋白(Munc18)抗体注射到大鼠的海马,大鼠呈痫样发作,故研究此抗体对海马神经细胞的损伤作用。方法取SD新生鼠海马组织,以此培养的原代海马神经细胞作为靶细胞。抗体在不同的作用时间(20h和40h)和稀释倍数下,通过细胞存活率和乳酸脱氢酶(LDH)释放量来测定抗体对神经细胞的毒性;脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(TUNEL)测定神经细胞的凋亡。结果Munc18抗体对神经细胞具有细胞毒性和促凋亡作用,并与抗体的滴度和作用时间呈剂量依赖性。即使在抗体低滴度的情况下(×1000和×2000),Munc18抗体仍可诱导凋亡,且与对照组比较有显著性差异(P<0.01或P<0.05)。结论Munc18抗体的致痫机制与其诱导神经细胞的损伤作用(可能主要是经凋亡途径)有关。Objective Primary study indicated that rat showed epileptic seizure on by injected anti-Munc18 antibody into hippocampus. So, this paper would investigate the effect of anti-Munc18 antibody induced injury of neuron in hippocampus. Methods Original hippocampal neurons, from hippocampal cortex of neonatal SD rat, were cultured as target cells. Anti-Munc18 antibody was carried out at two times window （20 h and 40 h） and at different dilutions. The survival of neurons and the release of lactate dehydrogenase were used to evaluate cytotoxicity, and TUNEL staining was applied to measure the apoptosis of nerve cell induced by anti-Munc18 antibody. Results Anti-Munc18 antibody could induce both cytotoxicity and apoptosis on original hippocampal neurons, which were concentration-dependent and time-dependent. Even though under low titer （ × 1 000 or × 2 000）, anti-Munc18 antibody could induce apoptosis, which was significantly higher than that in control group （ P 〈0. 01 or P 〈0. 05 ）. Conclusion The mechanism of anti-Munc18 antibody inducing epilepsy may be related to the injury of neuron with the main way of apoptosis.

关 键 词：突触前膜胞内蛋白 抗体 细胞毒性 凋亡 

分 类 号：R742.1[医药卫生—神经病学与精神病学]