丹酚酸B对VaD大鼠空间记忆及海马区PSD95突触相关蛋白影响  

Effects of salvianolic acid B on spatial memory and PSD95 synapse related protein in hippocampus of VaD rats

作　　者：李小楠 舒刚明[1] 高畅[1] 张云霞[1] 张莹[1] LI Xiaonan;SHU Gangming;GAO Chang;ZHANG Yunxia;ZHANG Yin(The Second Lead Department, The Fourth Medical Center, PLA General Hospital, Beijing 100081, China)

机构地区：[1]解放军总医院第四医学中心干二科,北京100081

出　　处：《西部医学》2022年第3期323-327,334,共6页Medical Journal of West China

摘　　要：目的探讨丹酚酸B(SalB)对血管性痴呆(VaD)大鼠海马神经元的保护作用及可能机制。方法60只雄性Wistar大鼠随机分为假手术组、模型组、治疗组及尼麦角林组4组,每组15只。建模成功后第7天,治疗组每天灌胃SalB 20 mg/kg、尼麦角林组每天灌胃麦角林7 mg/kg,假手术组和模型组给予同体积生理盐水,均连续治疗6周。采用水迷宫实验评价空间记忆力变化情况;酶联免疫吸附法(ELISA)检测乳酸脱氢酶(LDH)和丙二醛(MDA)含量,DCFH-DA法检测活性氧(ROS)含量;尼氏染色法观察海马区形态变化;透射电镜观察海马区超微结构;蛋白免疫印迹法(Western blot)检测海马区NR2A/B,PSD95、PSD93、p-CREB、t-CREB蛋白表达。结果水迷宫试验发现,SalB治疗可以逆转VaD大鼠的记忆障碍;模型组MDA、LDH和ROS水平均高于假手术组(P<0.01),经SalB治疗后,大鼠MDA、LDH和ROS水平均明显下降(P<0.05);尼氏染色观察显示,模型组海马区锥体细胞丢失明显,细胞层数和数量减少,而治疗组和尼麦角林组排列较为整齐,大部分细胞轮廓清晰;透射电镜观察发现,经SalB治疗后,大鼠海马区突触凹型、数量增加,活性带长度增加;Western blot检测显示,与假手术组比较,模型组PSD95、NR2A/B、PSD93蛋白水平及p-CREB/t-CREB比值均明显下降(P<0.01),而治疗组PSD95、NR2A/B、PSD93蛋白水平及p-CREB/t-CREB比值较模型组均明显上升(P<0.01)。结论丹酚酸B对血管性痴呆大鼠神经具有保护作用,其机制可能与减轻氧化应激和恢复突触蛋白有关。Objective To investigate the protective effect of SalB on hippocampal neurons in vascular dementia(VaD)rats.Methods 60 male Wistar rats were randomly divided into sham group,model group,treatment group and nemergolin group,with 15 rats in each group.The change of spatial memory was evaluated by water maze test.The LDH and MDA were detected by ELISA.The ROS detected by DCFH-DA.The morphological changes of hippocampus were observed by Nissl staining method.The ultrastructure of hippocampus was observed by transmission electron microscopy.The expression of NR2A/B,PSD95,PSD93,p-CREB and t-creb were detected by Western blot.Results Water maze test showed that SalB treatment could reverse the spatial memory disorders of model rats.The levels of MDA,LDH and ROS in CCH group were higher than those in sham group(P<0.01).After treatment with SalB,MDA,LDH and ROS levels decreased.Nissl staining showed that pyramidal cells in hippocampus of model group were lost obviously,and the number and number of cell layers were decreased.However,the arrangement of pyramidal cells in treatment group and nimergolin group was more orderly,and most of the cells were clear.Transmission electron microscopy showed that the number of synaptic pits and the length of active bands increased after SalB treatment.Western blotting results that compared with sham group,the levels of PSD95,NR2A/B,PSD93 protein and p-CREB/t-creb ratio in model group decreased(P<0.01).After treatment with SalB,the levels of PSD95,NR2A/B,PSD93 protein and p-CREB/t-creb ratio in treatment group increased(P<0.01).Conclusion SalB plays a neuro-protective role in VaD,which may be related to the reduction of oxidative stress and the recovery of synaptic protein.

关 键 词：丹酚酸B 大鼠 慢性脑缺血 血管性痴呆 氧化应激 

分 类 号：R969[医药卫生—药理学] R749.13[医药卫生—药学]