摘要
目的探讨酸枣仁-延胡索配伍对肝郁型失眠小鼠的镇静催眠作用及机制。方法(1)将96只昆明小鼠随机分为4组:正常组、酸枣仁组(2.25 g·kg^(-1))、延胡索组(1.50 g·kg^(-1))、酸枣仁-延胡索配伍组(简称配伍组,3.75 g·kg^(-1)),每组24只;每日1次,连续灌胃10 d,然后观察戊巴比妥钠阈下剂量(腹腔注射,35 mg·kg^(-1))诱导小鼠的睡眠发生率,阈上剂量(腹腔注射,45 mg·kg^(-1))诱导小鼠的睡眠潜伏期、睡眠时间。(2)将60只昆明小鼠随机分为5组:正常组、模型组、酸枣仁组(2.25 g·kg^(-1))、延胡索组(1.50 g·kg^(-1))、配伍组(3.75 g·kg^(-1)),每组12只。采用慢性夹尾刺激和对氯苯丙氨酸(PCPA,400 mg·kg^(-1))腹腔注射的复合因子造模法复制肝郁型失眠小鼠模型。每日1次,连续灌胃给药14 d。采用酶联免疫吸附(ELISA)法检测血清去甲肾上腺素(NE)、多巴胺(DA)、5-羟色胺(5-HT)的含量,检测垂体促肾上腺皮质激素(ACTH)及下丘脑促肾上腺皮质激素释放素(CRH)的含量;采用免疫组化法检测垂体ACTH和海马5-HT的蛋白表达;采用q RT-PCR法检测脑组织中K^(+)-Cl^(-)共转运体2(KCC2)、Na^(+),K^(+)-2Cl^(-)共转运体1(NKCC1)m RNA表达水平。结果(1)与正常组比较,给药组小鼠的睡眠发生率明显提高,睡眠时间显著延长(P<0.01);酸枣仁组及配伍组的睡眠潜伏期显著缩短(P<0.01)。(2)与正常组比较,模型组小鼠血清的NE、DA含量明显升高(P<0.05,P<0.01),5-HT含量明显降低(P<0.01);脑垂体ACTH及下丘脑CRH含量明显升高(P<0.01);垂体中ACTH的表达明显上调(P<0.01),海马中5-HT的表达明显下调(P<0.01);脑组织的KCC2 m RNA表达明显下调(P<0.01),NKCC1 m RNA表达明显上调(P<0.01)。与模型组比较,配伍组小鼠血清的NE、DA含量明显降低(P<0.01),5-HT含量明显升高(P<0.01);脑垂体ACTH及下丘脑CRH含量明显降低(P<0.01);脑垂体中ACTH的表达明显下调(P<0.01),海马中5-HT的表达明显上调(P<0.01);脑组织的KCC
Objective To explore the effect and mechanism of combination of Ziziphi spinosae semen and Corydalis rhizoma on insomnia mice model with liver depression syndrome.MethodsFirstly,ninety-six Kunming mice were randomly divided into 4 groups:normal group,Ziziphi spinosae semen group(2.25 g·kg^(-1)),Corydalis rhizoma group(1.50 g·kg^(-1))and compatibility group of Ziziphi spinosae semen and Corydalis rhizoma(compatibility group for short)(3.75 g·kg^(-1)).After 10 days of intragastric administration,we observed the sleep incidence of sodium pentobarbital(subthreshold doses)and sleep latency and prolonged asleep time(superthreshold doses).Secondly,sixty Kunming mice were randomly divided into 5 groups:normal group,model group,Ziziphi spinosae semen group(2.25 g·kg^(-1)),Corydalis rhizoma group(1.50 g·kg^(-1))and compatibility group(3.75 g·kg^(-1)),12 mice in each group.The mouse model of insomnia was established by chronic stimulation of tail clipping and intraperitoneal injection of PCPA(400 mg·kg^(-1)).After 14 days of intragastric administration,the contents of norepinephrine(NE),dopamine(DA)and 5-hydroxytryptamine(5-HT)in serum were detected by enzyme-linked immunosorbent assay(ELISA).The contents of pituitary corticotropin(ACTH)and hypothalamic corticotropin releasing hormone(CRH)were detected by the same method.The protein expressions of pituitary ACTH and hippocampal 5-HT were observed by immunohistochemistry,the expression of KCC2 m RNA and NKCC1 m RNA in brain tissue were detected by q RT-PCR technique.ResultsCompared with the normal group,the incidence of sleep in the treated group was higher,the sleep latency was significantly shortened and the sleep time was significantly prolonged(P<0.01).The contents of NE and DA in the serum of the model group were significantly increased(P<0.01,P<0.05),the contents of 5-HT was significantly decreased(P<0.01),ACTH and CRH in the brain tissue of the model group were significantly higher(P<0.01),the expression of ACTH was significantly increased(P<0.01),the expressio
作者
王婷婷
郝蕾
王芮
张一昕
王茜
刘温雨
WANG Tingting;HAO Lei;WANG Rui;ZHANG Yixin;WANG Xi;LIU Wenyu(Department of Pharmacy,Hebei University of Traditional Chinese Medicine,Hebei Higher Education Institute Applied Technology Research Center on TCM Formula Preparation,Shijiazhuang 050091 Hebei,China)
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2021年第5期619-626,共8页
Traditional Chinese Drug Research and Clinical Pharmacology
基金
国家自然科学基金项目(81503268)
中央财政中共卫生专项“2019年全国第四次中药资源普查项目——河北省(Z135080000022)”。
