熟地有效成分甘露三糖对高浓度皮质酮损伤的海马神经细胞SGK\BDNF\GCR表达的影响

Gene expression of SGK、BDNF、GCR following mannotr-iose of SHUDI treatment in hippocampal neurons with high dose corticosterone injury

目的:观察熟地有效成分甘露三糖对高浓度皮质酮诱导海马神经元损伤及学习记忆相关信号转导分子表达的影响。方法:用24小时新生大鼠原代培养海马神经细胞。第八天用药处理,将细胞分为5组:1:正常对照组;2:1×10-4M高浓度皮质酮模型组;3:10-4 M CORT+3nM RU38486拮抗剂组;4:10-4 M CORT+0.1g/L甘露三糖组;5:10-4 M CORT+40uM多奈哌齐阳性对照组。24h后,通过SYTO13-PI双荧光染色观察各组细胞的形态及存活情况,MTT法测定细胞活性,Western Blot检测学习记忆信号转导通路相关分子GCR、BDNF、SGK蛋白表达。结果:与正常组相比,模型组细胞死亡较多,细胞活力下降,GCR、BDNF、SGK蛋白表达均显著性降低;如上改变又均被皮质酮受体拮抗剂RU38486逆转;甘露三糖(0.1g/L)同阳性对照多奈派齐(40μM)均明显减少死细胞数量,提高细胞活力,提高模型细胞GCR、BDNF、SGK蛋白表达。结论:熟地有效成分甘露三糖保护大鼠海马神经细胞免遭高浓度皮质酮的损伤,通过调节学习记忆信号转导途径中的重要蛋白GCR、BDNF、SGK表达从而改善高浓度皮质酮致学习记忆功能退化。

Objective： To observe the effect of mannotriose in SHUDI on gene expression of the learning and memory related signaling molecules.in injured hippocampal neurons by high dose of corticosterone. Methods： Hippocampal neurons prime cultured were prepared from newborn（24h） rats,after 7 d incubation,the cultured neurons were treated with drug and divided into 5 groups.1： blank group;2：model group containing10-4M CORT;3：RU38486 group containing 10-4M CORT＋3nM RU38486;4：mannotriose group containing10-4M CORT＋0.1g/L mannotriose;5：positive control group containing10-4M CORT＋40μM aricept.48 hours later,SYTO13-PI double-fluorescence staining、MTT assay and Western Blot was carried out. Results：Compared with the control group,both the cell survival rate by SYTO13-PI double-fluorescence staining and cell viability by MTT decreased markedly in model group（P0.05）,GCR,BDNF and SGK protein expression significantly decreased.Mannotriose as Aricept and RU38486 as increased the above indices to varying degrees. Conclusion： 10-4M CORT had injured the hippocampus neurons.Mannotriose of SHUDI as a positive control aricept as could significantly prevent the hippocampus neuron from injury,probably retarding learning and memory functional deterioration induced by high dose of CORT.The regulatory mechanism maybe is that through signal transdu-ction pathway on learning and memory the mannotriose change the learning and memory-related signal transduction molecules gene expression such as GCR,BDNF and SGK.