中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2013年
11期
1229-1234
,共6页
蒋雷%王军%高蓉%肖杭%张劲松
蔣雷%王軍%高蓉%肖杭%張勁鬆
장뢰%왕군%고용%초항%장경송
甲基苯丙胺%4-AP敏感型钾通道%TEA敏感型钾通道%细胞凋亡%海马神经元
甲基苯丙胺%4-AP敏感型鉀通道%TEA敏感型鉀通道%細胞凋亡%海馬神經元
갑기분병알%4-AP민감형갑통도%TEA민감형갑통도%세포조망%해마신경원
Methamphetamine%4-AP sensitive K+ channel%TEA sensitive K+ channel%Cell apoptosis%Hippocampal neurons
目的 探讨甲基苯丙胺(Meth)对外向钾电流的影响及外向型钾通道在Meth引起的海马神经元损伤过程中的作用.方法 以分离出的怀孕18 d Sprague-Dawley大鼠胎鼠的海马神经元作为实验对象,分为对照组和Meth处理组,采用全细胞膜片钳的实验方法,分别记录Meth处理后外向4-AP和TEA敏感型钾电流大小的变化.通过MTT和TUNEL的方法,观察Meth作用后引起的电流变化对海马神经元活力和凋亡的影响.统计分析利用单因素方差分析结合LSD-t检验,以P＜0.05为差异具有统计学意义.结果 与对照组(n=10)比较[(87.4±12.5) pA/pF],Meth处理组(n=7)细胞外向4-AP敏感型钾电流密度明显增大[(120.1±19.6)pA/pF,P＜0.01],且呈剂量依赖性.MTT结果表明,Meth明显降低细胞的活力(48.72 ±4.38)％,与对照组(100.07±3.36)％比较,差异具有统计学意义,而钾通道抑制剂4-AP (61.39 ±3.15)％以及胞外高浓度K+溶液(78.25 ±9.42)％,能有效抑制Meth引起的细胞损伤.TUNEL实验进一步表明,Meth能引起海马神经元的凋亡,而4-AP和胞外高浓度K+溶液对Meth引起的细胞凋亡具有保护作用.结论 Meth引起外向4-AP敏感型钾电流增大,进而可能导致海马神经元的损伤.
目的 探討甲基苯丙胺(Meth)對外嚮鉀電流的影響及外嚮型鉀通道在Meth引起的海馬神經元損傷過程中的作用.方法 以分離齣的懷孕18 d Sprague-Dawley大鼠胎鼠的海馬神經元作為實驗對象,分為對照組和Meth處理組,採用全細胞膜片鉗的實驗方法,分彆記錄Meth處理後外嚮4-AP和TEA敏感型鉀電流大小的變化.通過MTT和TUNEL的方法,觀察Meth作用後引起的電流變化對海馬神經元活力和凋亡的影響.統計分析利用單因素方差分析結閤LSD-t檢驗,以P＜0.05為差異具有統計學意義.結果 與對照組(n=10)比較[(87.4±12.5) pA/pF],Meth處理組(n=7)細胞外嚮4-AP敏感型鉀電流密度明顯增大[(120.1±19.6)pA/pF,P＜0.01],且呈劑量依賴性.MTT結果錶明,Meth明顯降低細胞的活力(48.72 ±4.38)％,與對照組(100.07±3.36)％比較,差異具有統計學意義,而鉀通道抑製劑4-AP (61.39 ±3.15)％以及胞外高濃度K+溶液(78.25 ±9.42)％,能有效抑製Meth引起的細胞損傷.TUNEL實驗進一步錶明,Meth能引起海馬神經元的凋亡,而4-AP和胞外高濃度K+溶液對Meth引起的細胞凋亡具有保護作用.結論 Meth引起外嚮4-AP敏感型鉀電流增大,進而可能導緻海馬神經元的損傷.
목적 탐토갑기분병알(Meth)대외향갑전류적영향급외향형갑통도재Meth인기적해마신경원손상과정중적작용.방법 이분리출적부잉18 d Sprague-Dawley대서태서적해마신경원작위실험대상,분위대조조화Meth처리조,채용전세포막편겸적실험방법,분별기록Meth처리후외향4-AP화TEA민감형갑전류대소적변화.통과MTT화TUNEL적방법,관찰Meth작용후인기적전류변화대해마신경원활력화조망적영향.통계분석이용단인소방차분석결합LSD-t검험,이P＜0.05위차이구유통계학의의.결과 여대조조(n=10)비교[(87.4±12.5) pA/pF],Meth처리조(n=7)세포외향4-AP민감형갑전류밀도명현증대[(120.1±19.6)pA/pF,P＜0.01],차정제량의뢰성.MTT결과표명,Meth명현강저세포적활력(48.72 ±4.38)％,여대조조(100.07±3.36)％비교,차이구유통계학의의,이갑통도억제제4-AP (61.39 ±3.15)％이급포외고농도K+용액(78.25 ±9.42)％,능유효억제Meth인기적세포손상.TUNEL실험진일보표명,Meth능인기해마신경원적조망,이4-AP화포외고농도K+용액대Meth인기적세포조망구유보호작용.결론 Meth인기외향4-AP민감형갑전류증대,진이가능도치해마신경원적손상.
Objective To investigate the effects of methamphetamine (Meth) on the outward K+ currents and elucidate the role of outward K+ channels in Meth induced hippocampal neuron damage.Methods Hippocampal neurons were harvest from 18-day-old embryonic rats and were divided into two groups:the control group and the Meth treated group.Both of 4-AP and TEA sensitive K+ currents were recorded after the treatment of Meth by performing the whole cell patch clamp.Furthermore,the MTT and TUNEL assays were performed to evaluate the effects of K+ channel on hippocampal neuron damage mediated by Meth.For statistical comparison,One-way ANOVA and LSD multiple comparison test or t-test was used.P-value ＜ 0.05 was considered to be statistically significant.Results The density of 4-AP sensitive K+ channel currents in Meth treated group [(120.1 ± 19.6) pA/pF,n =7] were significantly increased when compared with control group [(87.4 ± 12.5) pA/pF,n =10,P ＜0.01] and the increments of the currents induced by Meth was dose dependent.The MTT data showed that the cell viability was obviously decreased in Meth treated group (48.72 ± 4.38) ％ relative to the control group (100.07 ± 3.36) ％.Moreover,application of K+ channel antagonist,4-AP (61.39 ± 3.15)％,and the high K+ solution (78.25 ± 9.42) ％ substantially enhanced the cell viability.The TUNEL assay showed there were protective effects of 4-AP and the high K+ solution against neuron damage observed during cells exposed to Meth.Conclusions The increments of 4-AP sensitive K+ channel currents induced by Meth might be involved in hippocampal neuron damage.
