Jiaotai Pill (交泰丸) Alleviates Insomnia through Regulating Monoamine and Organic Cation Transporters in Rats

Objective To reveal the effect and mechanism of Jiaotai Pill(交泰丸,JTP)on insomniac rats.Methods The insomniac model was established by intraperitoneal injection of p-chlorophenylalanine(PCPA).In behavioral experiments,rats were divided into control,insomniac model,JTP[3.3 g/(kg•d)],and diazepam[4 mg/(kg•d)]groups.The treatment effect of JTP was evaluated by weight measurement(increasement of body weight),open field test(number of crossings)and forced swimming test(immobility time).A high performance liquid chromatography-electrochemical detection(HPLC-ECD)method was built to determine the concentration of monoamine transmitters in hypothalamus and peripheral organs from normal,model,JTP,citalopram[30 mg/(kg•d)],maprotiline[40 mg/(kg•d)]and bupropion[40 mg/(kg•d)]groups.Expressions of serotonin transporter(SERT),dopamine transporter(DAT),and norepinephrine transporter(NET)were analyzed by quantitative polymerase chain reaction(qPCR)and Western blot in normal,model and JTP groups.A high performance liquid chromatography-electrospray ionization mass spectrometry(HPLC-ESI-MS/MS)method was established to determine the pharmacokinetics,urine cumulative excretion of metformin in vivo,and tissue slice uptake in vitro,which were applied to assess the activity of organic cation transporters(OCTs)in hypothalamus and peripheral organs.Results Compared with the insomniac model group,the body weight and spontaneous locomotor were increased,and the immobility time was decreased after treatment with JTP(P<0.01).Both serotonin and dopamine contents in hypothalamus and peripheral organs were increased(P<0.01).The norepinephrine content was increased in peripheral organs and decreased in hypothalamus(P<0.05 or P<0.01).At the same time,SERT,DAT,OCT1,OCT2,and OCT3 were down-regulated in hypothalamus and peripheral organs(P<0.05).NET was down-regulated in peripheral organs and up-regulated in hypothalamus(P<0.05 or P<0.01).Moreover,the activity of OCTs in hypothalamus and peripheral organs was inhibited(P<0.05).Conclusion JTP alleviates insomnia through regulation of monoaminergic system and OCTs in hypothalamus and peripheral organs.