益肾化浊方对阿尔茨海默病模型大鼠海马区Ca^(2+)浓度及钙相关记忆蛋白表达的影响  

Effects of Yishen Huazhuo Decoction on Expression of Ca^(2+) and Calcium Memory-related protein in the Hippocampus of Experimental Rat Model of Alzheimer's Disease

作　　者：李强[1] 王凯 孙伟明 郭威 徐家淳 毕聪聪 张玉莲[1] LI Qiang;WANG Kai;SUN Wei-ming;GUO Wei;XU Jia-chun;BI Cong-cong;ZHANG Yu-lian(Second Affiliated Hospital of Tianjin University of TCM, Tianfin 300150, China;Graduate School, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China)

机构地区：[1]天津中医药大学第二附属医院,天津300150 [2]天津中医药大学研究生院,天津300193

出　　处：《时珍国医国药》2018年第3期546-549,共4页Lishizhen Medicine and Materia Medica Research

基　　金：高等学校博士学科点专项科研基金(20131210110012);国家自然科学基金(81273940);国家中医药管理局中医药标准化项目[SATM-2015-BZ(273)];天津市科技计划项目(15ZXLCSY00020)

摘　　要：目的观察益肾化浊方对AD模型大鼠海马区Ca^(2+)浓度及钙相关记忆蛋白表达的影响,初步探讨益肾化浊方治疗AD的部分作用机制。方法将30只雄性SD大鼠随机分为假手术组,模型组和益肾化浊方组。采用右侧侧脑室注射Aβ25-35建立AD模型,造模成功后益肾化浊方组大鼠按照4.32g·kg^(-1)·d^(-1)剂量给予药液灌胃,假手术组和模型组给予等体积蒸馏水灌胃,持续4周。采用Rhod 2-AM荧光探针法和Western blot检测各组大鼠海马区突触Ca^(2+)浓度及Ca MKⅡ、NR2B、CREB、BDNF和Trk B蛋白的表达。结果 (1)Rhod 2-AM荧光探针法检测结果显示:与假手术组比较,模型组大鼠钙离子浓度显著升高(P<0.01);与模型组相比,益肾化浊方组钙离子浓度显著减低(P<0.01),差异具有统计学意义。(2)Western blot检测结果显示:与假手术组比较,模型组大鼠海马区NR2B、Ca MKⅡ、CREB、BDNF和Trk B蛋白的表达显著降低(P<0.01);与模型组比较,益肾化浊方组NR2B、Ca MKⅡ、CREB、BDNF和Trk B蛋白显著升高(P<0.01),差异具有统计学意义。结论益肾化浊方治疗AD的作用机制可能与调节海马突触体内Ca^(2+)浓度,减轻Aβ诱发的神经毒性;同时增加钙相关记忆蛋白的表达有关。Objective To observe the effect of Yishen Huazhuo Decoction（YHD） on expression of Ca^2＋ and calcium memory - related protein in the hippocampus of experimental rat model of Alzheimer＇s Disease and explore the mechanism of YHD for treating Alzheimer＇s disease. Methods Male SD rats were randomly divided into 3 groups, sham operation group, model group and YHD group. In this study, Aβ25-35 was injected into rat right paracele to establish AD model; Then the YHD group was given YHD （4.32g·kg^-1） within the next 4 weeks, 1 time a day. The sham operation group and model group were given the same volume of distilled water. Rhod 2 - AM fluorescent probe method and Western blot were used to detect the effect of YHD on concentrations of Ca^2＋ of synaptic and NR2B, CaMK Ⅱ , CREB, BDNF and TrkB in the hippocampus. Results （ 1 ） Rhod 2 - AM fluorescent probe assay showed： Compared with sham operation group, Ca^2＋ concentration of synaptosomes in hippocampus was significantly increased in AD rat of model group （P 〈 0.01 ） ; compared with the model group, Ca^2＋ concentration of synaptosomes in hippocampus was significantly decreased in AD rat of YHD group（ P 〈 0.01 ）. （ 2 ） Western blot test results showed ： Compared with the sham operation group, NR2B, CaMK I , CREB, BDNF and TrkB protein expression of AD rats in model group was significantly reduced（ P 〈 0.01 ） ; compared with the model group, YHD can obviously increase the expression of NR2B, CaMK Ⅱ , CREB, BDNF and TrkB（P 〈 0.01 ）. Conclusion The mechanism of the YHD treatment of AD, which might function in regulate the Ca^2＋ of synaptosomes and expression memory related protein, thereby reducing Aβ- induced neurotoxicity.

关 键 词：阿尔茨海默病 益肾化浊方 突触体 CA^2+ 钙相关记忆蛋白 

分 类 号：R285.5[医药卫生—中药学]