人参-五味子药对对PCPA失眠大鼠神经递质及cAMP/Epac/Raf1信号通路的作用机制研究  

Study on the Mechanisms of Renshen(Ginseng Radix Et Rhizoma)-Wuweizi(Schisandrae Chinensis Fructus)Drug Pair on the Neurotransmitter and cAMP/Epac/Raf1 Signalling Pathway of PCPA Insomnia Mice

作　　者：苏贺 乔铁[1] 梁可[1] 郑冰元[1] 李芳潇 SU He;QIAO Tie;LIANG Ke;ZHENG Bingyuan;LI Fangxiao(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China)

机构地区：[1]辽宁中医药大学,辽宁沈阳110847

出　　处：《辽宁中医药大学学报》2024年第10期20-24,共5页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：辽宁省教育厅科学研究项目(L202014);辽宁省高等学校创新人才支持计划项目(LR2019039);辽宁省重点实验室和技术创新技术中心组建计划项目(辽科发[2021]37号)。

摘　　要：目的通过观察人参-五味子药对对失眠大鼠下丘脑中神经递质及其cAMP/Epac/Raf1信号通路水平的影响,探讨人参-五味子药对对PCPA失眠大鼠的作用机制。方法将60只SD大鼠随机分为空白组、模型组、地西泮组、人参-五味子低剂量组、中剂量组和高剂量组,每组10只。除空白组外,其余5组连续3 d对大鼠进行腹腔注射对氯苯丙氨酸(PCPA)复制失眠模型。模型复制成功后,人参-五味子药对低、中、高剂量组给药浓度分别为0.3、0.9、2.7 g/mL,地西泮组给药浓度为0.002 g/mL,模型组、空白组每天生理盐水灌胃,各组每日灌胃2 mL,连续给药14 d。观察记录各组大鼠的行为学变化;采用HE(苏木素-伊红)染色法检测大鼠下丘脑组织病理形态的改变;ELISA(酶联免疫吸附法)测定大鼠下丘脑中甘丙肽(GAL)、去甲肾上腺素(NE)、5-羟色胺(5-HT)、腺苷(Ado)水平;RT-PCR(实时荧光定量聚合酶链式反应)检测大鼠下丘脑中Eapc、Raf1mRNA的表达;IHC(免疫组织化学染色法)检测大鼠下丘脑中cAMP蛋白的含量。结果和空白组相比:模型组大鼠下丘脑病理切片损伤明显;GAL、5-HT、Ado含量显著下降(P<0.05),NE含量显著上升(P<0.05),下丘脑Eapc、Raf1 mRNA表达显著下降(P<0.05),下丘脑cAMP蛋白表达显著下降(P<0.05),差异有统计学意义。与模型组相比:地西泮组、人参-五味子低、中、高剂量组下丘脑病理切片损伤明显改善;地西泮组、人参-五味子中、高剂量组GAL、5-HT、Ado含量显著上升(P<0.05),NE含量显著下降(P<0.05),下丘脑Eapc、Raf1 mRNA表达显著上升(P<0.05),下丘脑cAMP蛋白表达显著上升(P<0.05),差异有统计学意义。结论人参-五味子药对可能通过上调cAMP/Epac/Raf1信号通路,缓解PCPA大鼠失眠症状。Objective By observing the effects of Renshen(Ginseng Radix Et Rhizoma)-Wuweizi(Schisandrae Chinensis Fructus)drug pair on the levels of hypothalamic neurotransmitter and cAMP/Epac/Raf1 signaling pathway in insomnia rats,the mechanism of Renshen(Ginseng Radix Et Rhizoma)-Wuweizi(Schisandrae Chinensis Fructus)drug pair on insomnia rats with PCPA was investigated.Methods Sixty SD rats were randomly divided into blank group,model group,diazepam group,Renshen(Ginseng Radix Et Rhizoma)-Wuweizi(Schisandrae Chinensis Fructus)drug pair low-dose group,medium-dose group and high-dose group,with 10 rats in each group.Excepting the blank group,the other five groups were given peritoneal injection of p-chlorophenylalanine(PCPA)for three consecutive days to reproduce insomnia model.After the successful replication of the model,the concentrations of Renshen(Ginseng Radix Et Rhizoma)-Wuweizi(Schisandrae Chinensis Fructus)drug pair were 0.3,0.9 and 2.7 g/mL in the low,medium and high dose groups,respectively,and 0.002 g/mL in the diazepam group.The model group,the vacuum group injected normal saline into the stomach every day,each group infused 2 mL per day,continuously administered for 14 days.Observing behavioral changes in the groups of rats;using the Hematoxylin and Eosin staining(HE)method to detect changes in pathological patology of the subcervical tissue of the rats;Enzyme-connected immunosuppression method(ELISA)to determine the levels of galanin(GAL),norepinephrine(NE),5-hydroxytryptamine(5-HT),adenosine(Ado);real-time fluorescence quantitative polymerase chain reaction(RT-PCR)to detect the expression of Eapc,Raf1 mRNA in the hypothalamus of rats;The content of cAMP protein in rat hypothalamus was detected by immunohistochemical staining(IHC).Results Compared with the blank group:In the model group,the hypothalamic pathological sections of rats were significantly damaged;the contents of GAL,5-HT and Ado were significantly decreased(P<0.05),the content of NE was significantly increased(P<0.05),the mRNA expressions of Eap

关 键 词：人参-五味子药对 cAMP/Epac/Raf1信号通路 失眠 作用机制 

分 类 号：R285.5[医药卫生—中药学]