CAJ | 학술논문

目的观察补阳还五汤对血管性痴呆(vascular dementia,VD)大鼠海马N-甲基-D-门冬氨酸受体亚单位2B(N-Methyl-D-asparate receptor subunit 2B,NR2B)蛋白及其mRNA表达的影响,探讨补阳还五汤治疗VD的机制.方法四血管阻断(four vessel occlusion,4VO)法制备VD大鼠模型.设立假手术组、VD模型组、尼莫地平组(20mg·kg-1·d-1,灌胃30d)和补阳还五汤治疗组(50g生药·kg-1·d-1,灌胃30d).Morris水迷宫检测大鼠学习记忆能力的改变,免疫组化和Western-Blot技术检测大鼠海马神经元NR2B蛋白的变化,实时荧光定量PCR技术检测大鼠海马NR2B mRNA表达的变化.结果与假手术组大鼠逃避潜伏期[(24.18±7.90)s]和平均探索次数[(7.99±1.32)次/min]相比,VD模型组大鼠逃避潜伏期[(51.25±18.28)s]延长和平均探索次数[(5.26±0.74)次/min]减少(P<0.05);补阳还五汤组[(25.91±9.56)s和(7.52±1.27)次/min]明显改善了模型大鼠学习、记忆成绩(P<0.05);假手术组、尼莫地平组与补阳还五汤治疗组之间差异无显著性(P>0.05).VD模型组大鼠的NR2B蛋白(0.33±0.06)及其mRNA(593±53)表达水平较假手术组(0.71 ±0.13)、(5887±501)明显降低(P<0.05);补阳还五汤治疗组大鼠海马的NR2B蛋白(0.66±0.11)及其mRNA(5692±482)表达水平较模型组的表达明显升高(P<0.05);假手术组、尼莫地平组组与补阳还五汤治疗组之间差异无显著性(P>0.05).结论补阳还五汤可以改善VD大鼠学习记忆能力,其机制可能与减轻脑缺血再灌注对海马CAI区神经元的损伤及促进海马组织NR2B蛋白及其mRNA的表达有关.
목적 관찰보양환오탕대혈관성치태(vascular dementia,VD)대서해마N-갑기-D-문동안산수체아단위2B(N-Methyl-D-asparate receptor subunit 2B,NR2B)단백급기mRNA표체적영향,탐토보양환오탕치료VD적궤제.방법 사혈관조단(four vessel occlusion,4VO)법제비VD대서모형.설립가수술조、VD모형조、니막지평조(20mg·kg-1·d-1,관위30d)화보양환오탕치료조(50g생약·kg-1·d-1,관위30d).Morris수미궁검측대서학습기억능력적개변,면역조화화Western-Blot기술검측대서해마신경원NR2B단백적변화,실시형광정량PCR기술검측대서해마NR2B mRNA표체적변화.결과 여가수술조대서도피잠복기[(24.18±7.90)s]화평균탐색차수[(7.99±1.32)차/min]상비,VD모형조대서도피잠복기[(51.25±18.28)s]연장화평균탐색차수[(5.26±0.74)차/min]감소(P<0.05);보양환오탕조[(25.91±9.56)s화(7.52±1.27)차/min]명현개선료모형대서학습、기억성적(P<0.05);가수술조、니막지평조여보양환오탕치료조지간차이무현저성(P>0.05).VD모형조대서적NR2B단백(0.33±0.06)급기mRNA(593±53)표체수평교가수술조(0.71 ±0.13)、(5887±501)명현강저(P<0.05);보양환오탕치료조대서해마적NR2B단백(0.66±0.11)급기mRNA(5692±482)표체수평교모형조적표체명현승고(P<0.05);가수술조、니막지평조조여보양환오탕치료조지간차이무현저성(P>0.05).결론 보양환오탕가이개선VD대서학습기억능력,기궤제가능여감경뇌결혈재관주대해마CAI구신경원적손상급촉진해마조직NR2B단백급기mRNA적표체유관.
Objective To observe the effect of buyanghuanwu decoction on expression of immunoreactive protein and mRNA of NMDA receptor 2B subunit in rats hippocampal with vascular dementia to investigate the mechanism of buyanghuanwu decoction. Methods One hundred and forty-four rats were randomly divided into 4 groups: sham-operated group, VD model group,nimodipine group and buyanghuanwu decoction treatment group. The rats models of vascular dementia were built up by four vessels occlusion method. VD rats were treated with in-tragastrical buyanghuanwu decoction suspension (50 pharmacognostic g·kg-1·d-1) and nimodipine suspension (20 mg·kg-1·d-1) for 30 days. The learning and memory abilities were evaluated by Morris water maze tests. The change of NR2B protein in hippocampal of each group of rats were measured with immunohistochemistry and Western blot techniques and the expression of NR2B mRNA in hippocampus were observed by real-time fluorescence quantitative PCR techniques. Results Water maze tests,compared with sham-operated group((24. 18 ± 7.90)s,(7.99 ±1.32)/min) ,the escape latency(51. 25 ±18.28)s to explore the extension and the average spatial probe number ((5. 26 ±0. 74)/min) reduced in VD model group (P < 0. 05). Buyanghuanwu decoction ((25.91 ±9.56)s,(7. 52 ± 1. 27)/min) had significantly improved the above-mentioned rat model of learning and memory performance (P<0.05). There was no significant difference among sham-operated group,nimodipine group and buyanghuanwu decoction treatment group (P>0. 05). Similarly,as compared rats with sham-operated group(0.71 ±0.13), (5887 ±501), the expression of NR2B protein (0. 33 ± 0. 06) and its mRNA(593 ±53) were apparently decreased in VD rats (P< 0.05). The expression of NR2B protein(0.66 ±0. 11) and its mRNA (5692 ±482) in neuron of hippocampus were increased by buyanghuanwu decoction compared with the model group (P < 0. 05), and no difference was discovered between sham operation group and nimodipine group (P > 0.05).Conclusions Buyanghuanwu decoction improves the learning and memory abilities in VD rats, the therapeutic mechanism was concerned with lessening the injury of neurons on CA1 field in hippocampus and promoted the expression of NR2B protein and its mRNA.