左、右归丸对肾虚质大鼠海马区MEK/ERK/CREB信号通路的影响
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摘要
目的观察左、右归丸对肾虚质大鼠海马区MEK/ERK/CREB信号通路的影响,探讨其治疗肾虚质学习记忆障碍的作用靶点。方法采用"猫吓鼠"方法,构造先天不足+后天惊恐的复合型肾虚质仔鼠,分为肾虚组、左、右归丸组,空白组产自正常孕鼠。实验组予左、右丸浓缩液灌胃,2次/d,连续4周,空白组和肾虚组予等体积生理盐水。Morris水迷宫测定大鼠学习记忆能力。酶联免疫吸附法(ELISA)检测MEK、ERK、CREB蛋白含量。结果①水迷宫实验:对比空白组,肾虚质组逃避潜伏期、总路程、首次穿越目标时间延长;目标区域停留时间减少;实验组明显改善。②测定蛋白含量实验:对比空白组,肾虚质组MEK、ERK、CREB蛋白表达下降;实验组蛋白表达均明显上调。实验组间无显著性差异。结论海马区MEK/ERK/CREB信号通路可能是左、右归丸治疗肾虚质学习记忆减退的作用靶点之一。
Objective To observe the effect of ZuoGui and YouGui Pill on MEK/ERK/CREB signaling pathway in hippocampus of rats with kidney deficiency, and to explore its target for treating learning and memory impairment of kidney deficiency. Methods By the way of "cat scare rat", a complex kidney deficiency model of the congenital deficiency + acquired panic is constructed. Rats in the ZuoGui Pill group and the YouGui Pill group are administered by gastrogavage with ZuoGui and YouGui suspension. Blank group and model group are given equal volume of normal saline by gastrogavage. The learning and memory ability of rats is determined by Morris water maze test. The content of MEK, ERK and CREB is detected by ELISA. Results In the Morris water maze experiment, compared with the blank group, the latency period, total distance, and time of firstcrossing the target area of model group are all significantly expanded and time in the target area are shortened. After drug intervention, the learning and memory ability of ZuoGui Pill group and YouGui Pill group are improved. In the ELISA experiment, The protein expressions of MEK, ERK and CREB in hippocamus are lower in the model group than in the blank control group. But when compared with the model group,they are obviously higher in the ZuoGui Pill group and YouGui Pill group. There is no significant difference between Zuogui Pill group and Yougui Pill group. Conclusion The signaling pathway of MEK/ERK/CREB in hippocampus is one of the target of ZuoGui and YouGui Pill to treat learning and memory decline of kidney deficiency.
Objective To observe the effect of ZuoGui and YouGui Pill on MEK/ERK/CREB signaling pathway in hippocampus of rats with kidney deficiency, and to explore its target for treating learning and memory impairment of kidney deficiency. Methods By the way of "cat scare rat", a complex kidney deficiency model of the congenital deficiency + acquired panic is constructed. Rats in the ZuoGui Pill group and the YouGui Pill group are administered by gastrogavage with ZuoGui and YouGui suspension. Blank group and model group are given equal volume of normal saline by gastrogavage. The learning and memory ability of rats is determined by Morris water maze test. The content of MEK, ERK and CREB is detected by ELISA. Results In the Morris water maze experiment, compared with the blank group, the latency period, total distance, and time of firstcrossing the target area of model group are all significantly expanded and time in the target area are shortened. After drug intervention, the learning and memory ability of ZuoGui Pill group and YouGui Pill group are improved. In the ELISA experiment, The protein expressions of MEK, ERK and CREB in hippocamus are lower in the model group than in the blank control group. But when compared with the model group,they are obviously higher in the ZuoGui Pill group and YouGui Pill group. There is no significant difference between Zuogui Pill group and Yougui Pill group. Conclusion The signaling pathway of MEK/ERK/CREB in hippocampus is one of the target of ZuoGui and YouGui Pill to treat learning and memory decline of kidney deficiency.
引文
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[12]颜晓红,李友筑,周卫东,等.密闭式体外培养系统对胚胎发育及小鼠学习和记忆能力的影响[J].生殖医学杂志,2018,27(10):1006-1010.
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[16]Yang B H,Son H,Kim S H,et al.Phosphorylation of ERK and CREBin cultured hippocampal neurons after haloperidol and risperidone administration[J].Psychiatry&ClinicalNeurosciences,2004,58(3):262-267.
[2]张登本.《黄帝内经》二十论[M].北京:中国中医药出版社,2017:130.
[3]李林,魏海峰,张兰,等.中医“肾生髓,脑为髓海”现代生物学基础探讨[J].中国中药杂志,2006(17):1397-1400+1417.
[4]孔明望,赵健邦,石和元,等.补肾化痰益智方对APPV717I小鼠学习记忆能力和海马神经元Tau蛋白磷酸化的影响[J].中国实验方剂学杂志,2017,23(12):96-102.
[5]李莹,刘爽.补肾中药治疗老年性痴呆研究进展[J].辽宁中医杂志,2016,43(6):1338-1340.
[6]尹芳,孙丽娥,康湘萍,等.补肾益气方药对老年大鼠空间学习记忆能力及相关蛋白表达的影响[J].时珍国医国药,2017,28(7):1574-1576.
[7]曹娟,董联玲,冀俊虎,等.红景天对老年性痴呆大鼠行为学及海马区凋亡相关蛋白表达的影响[J].中国中医急症,2008(11):1567-1569.
[8]孙理军,孙耀光,张琪,等.肾虚质大鼠生长发育和学习记忆能力与肾藏志理论相关性研究[J].中华中医药杂志,2016,31(1):36-38.
[9]孙瑜嬬,孙理军.左、右归丸对复合型肾虚体质大鼠神经行为干预的实验研究[J].时珍国医国药,2015,27(1):72-74.
[10]孙理军,李翠娟,王震,等.肾虚质实验动物模型的构建方法与评价[J].时珍国医国药,2013,24(1):247-249.
[11]施新猷.现代医学实验动物学[M].北京:人民军医出版社,2000:332-335.
[12]颜晓红,李友筑,周卫东,等.密闭式体外培养系统对胚胎发育及小鼠学习和记忆能力的影响[J].生殖医学杂志,2018,27(10):1006-1010.
[13]De Vasconcellos A P,Zugno A I,Dos Santos A H,et al.Na+,K+-ATPase activity is reduced in hippocampus of rats submittedto an experimental model of depression:effect of chronic lithium treatment and possible involvement in learning deficits[J].Neurobiol Learn Mem,2005,84(2):102.
[14]Silva A J,Frankland P W,Marowit Z,et al.A mouse model for the learning and memory deficits associated with neurofibromatosis typeI[J].Nature Genetics,1997,15(3):281-284.
[15]Hayward P.Presenilin dysfunction leads to memory and plasticity defects[J].Lancet Neurology,2004,3(6):327.
[16]Yang B H,Son H,Kim S H,et al.Phosphorylation of ERK and CREBin cultured hippocampal neurons after haloperidol and risperidone administration[J].Psychiatry&ClinicalNeurosciences,2004,58(3):262-267.