李强^1，， 王凯^2，， 孙伟明^2，， 郭威^2，， 徐家淳^2，， 毕聪聪^2，， 张玉莲^1，*

（  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；  ）

摘要： 目的：观察益肾化浊方对AD模型大鼠海马区Ca2+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4.32gokg-1od-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Ca2+浓度及CaMKⅡ、NR2B、CREB、BDNF和TrkB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、CaMKⅡ、CREB、BDNF和TrkB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、CaMKⅡ、CREB、BDNF和TrkB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Ca2+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。
关键词： 阿尔茨海默病；益肾化浊方；突触体；Ca2+；钙相关记忆蛋白?????

LI qiang^1,， WANG Kai^2,， SUN Weiming^2,， GUO Wei^2,， XU Jiachun^2,， BI Congcong^2,， ZHANG Yulian^1,*

（  1、The second affiliated hospital of tianjin university of TCM,Tianjin 300150；       2、Graduate school,Tianjin University of Traditional Chinese Medicine,Tianjin 300193；  ）

Abstract： Objective: To observe the effect of Yishen Huazhuo Decoction(YHD) on expression of Ca2+ 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 (4.32gokg-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 Ca2+ of synaptic and NR2B, CaMKⅡ, CREB, BDNF and TrkB in the hippocampus. Results: (1)Rhod 2-AM fluorescent probe assay showed: Compared with sham operation group, Ca2+ concentration of synaptosomes in hippocampus was significantly increased in AD rat of model group (p<0.01); compared with the model group, Ca2+ 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, CaMKⅡ, 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, CaMKⅡ, CREB, BDNF and TrkB(p<0.01). Conclusions: The mechanism of the YHD treatment of AD, which might function in regulate the Ca2+ of synaptosomes and expression memory related protein, thereby reducing Aβ-induced neurotoxicity.?????
Keywords： Alzheimer's disease, Yishen Huazhuo Decoction, synaptosomes, Ca2+, calcium memory-related protein?????

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