疏肝健脾养心方对失眠模型小鼠食欲素A及其受体的干预作用  

Shugan Jianpi Yangxin Prescription Regulates Orexin-A and Its Receptors in Mouse Model of Insomnia

作　　者：王震 张杰[2] 刘飞祥 孙江燕 高亚慧[1] 郑育新 赵敏 WANG Zhen;ZHANG Jie;LIU Feixiang;SUN Jiangyan;GAO Yahui;ZHENG Yuxin;ZHAO Min(First Clinical Medical College,Henan University of Chinese Medicine,Zhengzhou 450000,China;Encephalopathy Hospital,The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China)

机构地区：[1]河南中医药大学,第一临床医学院,郑州450000 [2]河南中医药大学第一附属医院,脑病医院,郑州450000

出　　处：《中国实验方剂学杂志》2024年第10期54-61,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家中医药传承创新专项(2022JDZX004,2022CCCX008)。

摘　　要：目的:探讨疏肝健脾养心方下调食欲素A(OXA)及食欲素受体1(OX1R)、食欲素受体2(OX2R)的表达对失眠模型小鼠的干预作用。方法:利用腹腔注射对氯苯丙氨酸(PCPA)建立失眠小鼠模型,50只BALB/c小鼠随机分为空白组、模型组、右佐匹克隆组(0.13 mg·kg^(-1))、疏肝健脾养心方低、高剂量组(8.4、33.6 g·kg^(-1)),并给予相应药物治疗14 d。监测小鼠体质量变化,分别进行Morris水迷宫、戊巴比妥钠协同睡眠实验;免疫组化(IHC)检测下丘脑OXA的表达;酶联免疫吸附测定法(ELISA)检测下丘脑、血清、脾脏组织中OXA、5-羟色胺(5-HT)含量;实时荧光定量聚合酶链式反应(Real-time PCR)检测下丘脑OXA及其受体OX1R、OX2R mRNA表达。结果:与空白组比较,模型组小鼠体质量显著降低(P<0.01),逃避潜伏期时间显著增加(P<0.01),睡眠潜伏期显著增加(P<0.01),睡眠持续时间显著减少(P<0.01),下丘脑、血清、脾脏组织中OXA含量升高、5-HT含量明显降低(P<0.05),下丘脑中OXA及其受体的mRNA含量显著升高(P<0.01)。与模型组比较,疏肝健脾养心方低、高剂量组小鼠体质量明显升高(P<0.05,P<0.01),逃避潜伏期时间明显减少(P<0.05),睡眠潜伏期减少,睡眠持续时间显著增加(P<0.01),下丘脑、血清、脾脏组织中OXA含量降低、5-HT含量明显升高(P<0.05,P<0.01),下丘脑中OXA及其受体的mRNA含量显著降低(P<0.01)。结论:疏肝健脾养心方具有镇静、催眠作用,对失眠症有一定的治疗作用,其机制与增加脑中5-HT的含量,抑制下丘脑中OXA及其受体的表达有关。Objective:To investigate the interventional effects of Shugan Jianpi Yangxin prescription on the expression of orexin-A(OXA),orexin-1 receptor(OX1R),and orexin-2 receptor(OX2R)in the mouse model of insomnia.Method:The mouse model of insomnia was established by intraperitoneal injection of DL-4-chlorophenylalanine(PCPA).Fifty BALB/c mice were randomized into a blank group,a model group,an eszopiclone(0.13 mg·kg^(-1))group,and low-and high-dose(8.4 and 33.6 g·kg^(-1),respectively)Shugan Jianpi Yangxin prescription groups and treated with the corresponding drugs for 14 consecutive days.The weight changes of mice were monitored,and Morris water maze and pentobarbital-induced sleep tests were conducted.Immunohistochemistry(IHC)was employed to examine the expression of OXA in the hypothalamus.Enzymelinked immunosorbent assay was used to measure the levels of OXA and 5-hydroxytryptamine(5-HT)in the hypothalamus,serum,and spleen.Real-time fluorescence quantitative polymerase chain reaction was employed to determine the mRNA levels of OXA,OX1R,and OX2R in the hypothalamus.Result:Compared with the blank group,the model group had decreased body weight(P<0.01),increased escape latency(P<0.01),increased sleep latency(P<0.01),shortened sleep duration(P<0.01),elevated OXA level and lowered 5-HT level in the hypothalamus,serum,and spleen(P<0.05),and up-regulated mRNA levels of OXA,OX1R,and OX2R in the hypothalamus(P<0.01).Compared with the model group,the low-and high-dose groups of Shugan Jianpi Yangxin prescription showed increased body weight(P<0.05,P<0.01),shortened escape latency(P<0.05),shortened sleep latency and prolonged sleep duration(P<0.01),and lowered OXA level and elevated 5-HT level in the hypothalamus,serum,and spleen(P<0.05,P<0.01).Moreover,the two doses of Shugan Jianpi Yangxin prescription down-regulated the mRNA levels of OXA,OX1R,and OX2R in the hypothalamus(P<0.01).Conclusion:Shugan Jianpi Yangxin prescription exerts sedative and hypnotic effects in mice by increasing the content of 5-HT in the brain and

关 键 词：疏肝健脾养心方 失眠 食欲素A(OXA) 5-羟色胺(5-HT) 

分 类 号：R2-0[医药卫生—中医学] R22R242R285.5