骨碎补总黄酮对记忆障碍模型小鼠ER-P38/MAPK通路的影响  

Effect of Rhizoma Drynariae total flavonoids on ER-P38/MAPK pathway of memory impairedmice

作　　者：赵德萍 雷霞[2] 佟玉良 崔悦 王媛 邢怀钰 孙慧峰[1] 张宁[1,2] Zhao Deping;Lei Xia;Tong Yuliang;Cui Yue;Wang Yuan;Xing Huaiyu;Sun Huifeng;Zhang Ning(College of Pharmacy,Heilongjiang University of Chinese Medicine,Harbin 150040,China;Jiamusi College,Heilongjiang University of Chinese Medicine,Jiamusi 154007,China;College of Pharmacy,Jiamusi University,Jiamusi 154007,China)

机构地区：[1]黑龙江中医药大学药学院,哈尔滨150040 [2]黑龙江中医药大学佳木斯学院,佳木斯154007 [3]佳木斯大学药学院,佳木斯154007

出　　处：《国际中医中药杂志》2022年第4期409-415,共7页International Journal of Traditional Chinese Medicine

基　　金：黑龙江省大学生创新创业训练计划(201910228083);哈尔滨市应用技术研究与开发项目(青年后备人才A类,2017RAQXZ113);黑龙江中医药大学博士创新基金(2017BS09);黑龙江中医药大学优秀创新人才支持计划(2018RCD19);黑龙江中医药大学研究生创新科研项目(2019yjscxo16)。

摘　　要：目的研究骨碎补总黄酮对亚硝酸钠诱导的学习记忆障碍模型小鼠学习记忆的影响。方法将75只小鼠按随机数字表法分为空白组、模型组、抗脑衰胶囊组、骨碎补总黄酮组和骨碎补总黄酮+抑制剂组,每组15只。抗脑衰胶囊组灌胃抗脑衰胶囊混悬液585 mg/kg;骨碎补总黄酮组灌胃骨碎补总黄酮溶液97.5 mg/kg;骨碎补总黄酮+抑制剂组腹腔注射0.072 mg/kg ICI182780溶液,并灌胃骨碎补总黄酮溶液97.5 mg/kg,连续给药21 d,1次/d。第22天进行造模,除空白组外,其余各组小鼠腹腔注射亚硝酸钠复制学习记忆障碍模型。采用水迷宫试验检测小鼠学习记忆能力,免疫组织化学染色检测小鼠海马雌激素受体β(ERβ)表达,Western blot法检测海马中ERβ、磷酸化p38(P-P38)、Bcl-2、Bcl-2关联死亡启动子(Bcl2 antagonist of cell death,Bad)、Caspase-3蛋白表达,比色法检测海马MDA、SOD、NO水平。结果与模型组比较,骨碎补总黄酮组小鼠潜伏期缩短(P<0.01),穿越平台次数及靶象限停留时间增加(P<0.01);小鼠海马区ERβ[(0.371±0.010)比(0.124±0.009)]、Bcl-2[(1.146±0.028)比(0.726±0.016)]蛋白表达及SOD[(153.657±6.385)U/mg比(67.719±5.845)U/mg]活性增加(P<0.01);P-P38/P38[(0.412±0.043)比(0.806±0.069)]、Bad[(0.421±0.010)比(0.633±0.010)]、Caspase-3[(0.923±0.042)比(1.437±0.033)]蛋白表达及MDA[(8.669±0.662)nmol/mg比(11.772±1.054)nmol/mg]、NO[(4.259±0.225)nmol/mg比(10.805±0.415)nmol/mg]水平降低(P<0.01)。ER阻断剂可拮抗骨碎补总黄酮的恢复和改善作用。结论骨碎补总黄酮可通过雌激素受体-P38丝裂原激活的蛋白激酶(ER-P38/MAPK)信号通路调节亚硝酸钠模型小鼠的学习记忆功能,抑制神经细胞凋亡,减少氧化应激损伤,从而发挥神经保护作用。Objective To study the effect of total flavonoids of Rhizoma Drynariae on learning and memory impairment mice induced by sodium nitrite.Methods 75 mice were divided into blank group,model group,Kangnaoshuai capsule group,Rhizoma Drynariae total flavonoids group and Rhizoma Drynariae total flavonoids+inhibitor group according to the random number table method,with 15 mice in each group.The Kangnaoshui Capsule group was administered with Kangnaoshui Capsule 585 mg/kg,the Rhizoma Drynariae total flavonoids group was administered with the Rhizoma Drynariae total flavonoids 97.5 mg/kg,the Rhizoma Drynariae total flavonoids group and the inhibitor group were administered with the Rhizoma Drynariae total flavonoids by intragastric administration 97.5 mg/kg,and intraperitoneal injection of 0.072 mg/kg ICI182780 for 21 days,once a day.The model was established on the 22nd day.Except for the blank group,the other mice were injected with sodium nitrite intraperitoneally to replicate the mice model with impaired learning and memory capability.The learning and memory capabilit of mice were detected with water maze method,and the estrogen receptor in hippocampus was detected by immunohistochemistryβ(estrogen receptorβ,ERβ).The expression of ERβin hippocampus and the expression of phosphorylated P38(P-P38)and the protein contents of B-cell lymphoma-2(Bcl-2),Bcl-2 associated death promoter(Bad)and Caspase-3 in the apoptotic system was detected by Western blot.The kit was used to detect MDA,SOD and NO protein content in hippocampus.Results The latency of Rhizoma Drynariae total flavonoids group was significantly shorter than the model group,the number of crossing platform and the residence time in the target quadrant were significantly increased(P<0.01);The expression of ERβProtein in mice hippocampus(0.371±0.010 vs.0.124±0.009),Bcl-2 protein(1.146±0.028 vs.0.726±0.016)and the contents of SOD[(153.657±6.385)U/mg vs.(67.719±5.845)U/mg]increased significantly(P<0.01);The expression of P-P38/P38 protein(0.412±0.043 vs.0.

关 键 词：记忆障碍 骨碎补总黄酮 亚硝酸钠 植物雌激素 细胞凋亡 氧化性应激 小鼠 

分 类 号：R285.5[医药卫生—中药学] R-332[医药卫生—中医学]