摘要
目的了解孕期边缘型维生素A(vitamin A,VitA)缺乏对氮-甲基-D-天冬氨酸(N-Methyl-D-Aspartate,NMDA)诱导的新生大鼠海马神经元Ca2+内流的影响。方法将16只健康雌性Wistar大鼠采用随机数字表法分为边缘型VitA缺乏(MVAD)和VitA正常(VAN)组,每组8只。从交配前3周开始分别饲以VitA缺乏饲料(含VitA400 IU/kg)和VitA充足饲料(含VitA6 500 IU/kg)。高效液相技术检测血清VitA浓度。取MVAD组及VAN组新生大鼠海马神经元进行原代培养,并运用钙影像测定仪检测原代培养的海马神经元在NMDA刺激后细胞内钙离子变化。Western blot半定量分析海马组织NMDA受体-1(NR1)的表达。结果 MVAD组母鼠血清VitA水平[(0.768±0.069)μmol/L]低于VAN组[(1.284±0.217)μmol/L],MVAD组仔鼠血清VitA水平[(0.806±0.092)μmol/L]低于VAN组[(1.270±0.098)μmol/L](P<0.01);神经元特异性烯醇化酶(neuron-specific enolage,NSE)免疫荧光鉴定神经元纯度显示两组无显著差异(P>0.05);MVAD组与VAN组海马神经元在NMDA刺激后,MVAD组Ratio340/380增加值(0.136±0.012)低于VAN组(0.168±0.022)(P<0.01);Western blot半定量分析结果显示NR1在MVAD组海马组织中的表达显著低于VAN组。结论孕期开始的MVAD会抑制NMDA诱导的新生大鼠海马神经元钙离子内流,而降低NR1在神经元的表达可能是其损伤的机制之一。
Objective To study the effect of vitamin A deficiency during pregnancy on N-methyl-D- aspartate （NMDA）-induced Ca2＋ influx of hippocampal neurons in newborn rats. Methods Sixteen healthy male Wistar rats were randomly divided into vitamin A deficiency group （ n = 8 ） and normal vitamin A group （n = 8 ） , and fed with vitamin A deficient breed and vitamin A sufficient breed, respectively, 3 weeks before copulation. Serum vitamin A level was measured by high performance liquid chromatography. Changes of Ca2＋ in primarily cultured hippocampal neurons of newborn rats were detected with a calcium imaging detector after treatment with NMDA. Expression of NMDA receptor 1 in hippocampus was detected by Western blotting. Results The serum vitamin A level was lower in vitamin A deficient group than in vitamin A sufficient group of mother and newborn rats [ （0. 768± 0. 069 ） μmol/L and （ 0. 806 ± 0. 092 ） μmoL/L vs （ 1. 284 ± 0. 217 ） μmol/L and （ 1. 270± 0. 098 ）μmol-/L, P 〈 0.01 ]. Immunofluorescence showed no significant difference in neuron-specific enolase （NSE） , between the 2 groups. The ratio of hippocampal neurons was lower in vitamin A deficient group than in vitamin A sufficient group after treatment with NMDA [ （0. 136 ±0.012） vs （0. 168±0. 022） , P 〈0.01 ]. Western blot analysis showed that the expression level of NMDA receptor 1 was significantly lower in vitamin A deficient group than in vitamin A sufficient group[ （2. 527±0.245 ） vs （ 1. 157 ±0. 197） ,P 〈 0. 05]. Conclusion Vitamin A deficiency during pregnancy inhibits Ca2＋ influx of hippocampal neurons in newborn rats by decreasing the expression of NMDA receptor 1 in the hippoeampus, which may be one of the mechanisms underlying cell damage.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2010年第15期1613-1616,共4页
Journal of Third Military Medical University
基金
国家自然科学基金(30600495)~~
