Comparative brain pharmacokinetic study of Jiaotai Pills in normal and insomnic rats using brain microdialysis combinated with LC–MS/MS

Objective：To compare the brain pharmacokinetics of five protoberberine-type alkaloids（i.e.berberine,palmatine,coptisine,epiberberine,and jatrorrhizine）,which were the main bioactive constituents of Jiaotai Pills（JTP）,in normal and insomnic rats orally administrated with JTP.Methods：The detection was conducted by a fully validated liquid chromatography-tandem mass spectrometry combinated with brain microdialysis method.Brain microdialysis probes were inserted into the hippocampus of rats.JTP extracts were administrated intragastrically and then brain microdialysates were collected at 30 min time intervals for 10 h.The separation of the five protoberberine-type alkaloids was carried out on a BDS Hypersilusing a mobile phase consisting of acetonitrile and water（containing 5 mmol ammonium acetate adjusted to p H 5.0）within 4 min.The quantification was performed by multiple reaction monitoring with the transitions of m/z 336.0-320.1 for berberine,m/z 352.0-336.1for palmatine,m/z 338.0-322.1 for jatrorrhizine,m/z 336.0-320.1 for epiberberine,m/z 320.0-292.1 for coptisine and m/z 356.4-192.1 for IS.Results：The lower limit of quantification for five protoberberine-type alkaloids was 0.05 ng/m L.Linearity,accuracy,precision,stability and matrix effect of five analytes were all satisfactory.Five protoberberinetype alkaloids were quickly distributed in the brain.Moreover,significant differences in the principal pharmacokinetic parameters such as AUC andthe analytes were observed between two groups.Conclusion：The LC-MS/MS method combinated with microdialysis is useful in the brain pharmacokinetic study of five protoberberine-type alkaloids.The results indicated that the rates of analytes absorption in insomnic rats were significantly higher than those in normal rats.Besides,the protoberberine-type alkaloids could bring a direct effect on the neuron in the hippocampus.

Molecular targets and mechanisms of Jiawei Jiaotai Pill on diabetic cardiomyopathy based on network pharmacology

BACKGROUND Jiawei Jiaotai Pill is commonly used in clinical practice to reduce apoptosis,increase insulin secretion,and improve blood glucose tolerance.However,its mechanism of action in the treatment of diabetic cardiomyopathy(DCM)remains unclear,hindering research efforts aimed at developing drugs specifically for the treatment of DCM.AIM To explore the pharmacodynamic basis and molecular mechanism of Jiawei Jiaotai Pill in DCM treatment.METHODS We explored various databases and software,including the Traditional Chinese Medicine Systems Pharmacology Database,Uniport,PubChem,GenCards,String,and Cytoscape,to identify the active components and targets of Jiawei Jiaotai Pill,and the disease targets in DCM.Protein-protein interaction network,gene ontology,and Kyoto Encyclopedia of Genes and Genomes analyses were used to determine the mechanism of action of Jiawei Jiaotai Pill in treating DCM.Molecular docking of key active components and core targets was verified using AutoDock software.RESULTS Total 42 active ingredients and 142 potential targets of Jiawei Jiaotai Pill were identified.There were 100 common targets between the DCM and Jiawei Jiaotai Pills.Through this screening process,TNF,IL6,TP53,EGFR,INS,and other important targets were identified.These targets are mainly involved in the positive regulation of the mitogen-activated protein kinase(MAPK)MAPK cascade,response to xenobiotic stimuli,response to hypoxia,positive regulation of gene expression,positive regulation of cell proliferation,negative regulation of the apoptotic process,and other biological processes.It was mainly enriched in the AGE-RAGE signaling pathway in diabetic complications,DCM,PI3K-Akt,interleukin-17,and MAPK signaling pathways.Molecular docking results showed that Jiawei Jiaotai Pill's active ingredients had good docking activity with DCM's core target.CONCLUSION The active components of Jiawei Jiaotai Pill may play a role in the treatment of DCM by reducing oxidative stress,cardiomyocyte apoptosis and fibrosis,and maintaining metabolic homeostasis.

Optimization ratio between Coptis chinensis and cinnamo of Jiaotai pill plus benzodiazepines for insomnia in China:a network meta-analysis

OBJECTIVE To explore the efficacy of Jiaotai pill(JTW)and JTW modified prescriptions(JTW^(+))plus benzodiazepines(BDZ)for insomnia in China by systematic review and network meta-analysis,especially optimization ratio(_(m:n))between Coptis chinensis and Cinnamo.METHODS Randomized controlled trials(RCTs)regarding JTW and JTW^(+)+BDZ for insomina were comprehensively searched in CNKI,VIP,Wanfang,and PubMed,Cochrane Library,OVID,Embase databases from their inception to December 2017.The risk of bias was assessed by the Cochrane collaboration′s tool and Jadad scale.Systematic review and network meta-analysis were performed to evaluate the relative risk(RR)plus 95%confidence interval(95%CI)or prediction interval(95%PrI),and draw surface under the cumulative ranking curves(SUCRA)by Stata 14.0 and RevMan 5.0 software.RESULTS A total of 13 RCTs(1 RCT was omitted by sensitivity analysis)with 1040 patients were included.Overall heterogeneity was acceptable,except publication bias.Based on classical meta-analysis,JTW and JTW_(m:n)^(+)+BDZ,such as JTW_(2:1)^(+)+BDZ,JTW_(5:1)^(+)+BDZ,JTW_(6:1)^(+)+BDZ,JTW_(10:1)^(+)+BDZ,had more efficacy than BDZ.Based on network meta-analysis,JTW and JTW_(m:n)^(+)+BDZ,such as JTW_(2:1)^(+)+BDZ,JTW_(6:1)^(+)+BDZ,JTW_(10:1)^(+)+BDZ,had more efficacy than BDZ;meanwhile,SUCRA showed the best rank of JTW/JTW+(BDZ<JTW_(m:n)^(+)+BDZ<JTW)for insomina,as same as that of JTW_(m:n)+(BDZ<JTW_(5:1)^(+)+BDZ<JTW_(10:1)^(+)+BDZ<JTW_(2:1)^(+)+BDZ<JTW_(6:1)^(+)+BDZ).CONCLUSION Based on the existing clinical evidence,JTW has efficacy for insomnia,but the optimization ratio between Coptis chinensis and cinnamo of JTW_(m:n)^(+)+BDZ for insomnia needs more supports.

Pharmacokinetics of Jiaotai pill self-microemulsion in insomnia rats

Objective:The pharmacokinetics and relative bioavailability of Jiaotai pill self-microemulsion were evaluated by investigating the blood concentration of Berberine,Coptisine,Palmatine and Jatrorrhizine in insomnia rats.Methods:Insomnia rat model was established by intraperitoneal injection of p-chlorophenylalanine(PCPA).The model rats were given Jiaotai pill self-microemulsion and Jiaotai pill suspension.The contents of Berberine,Coptisine,Palmatine and Jatrorrhizine in plasma at different times after administration were determined by UPLC-MS/MS,and calculate pharmacokinetic parameters.Results:Under the set chromatographic conditions,the linear relationship of the four components was good,and the precision,accuracy and stability meet the requirements of biological samples.After intragastric administration of Jiaotai pill self-microemulsion,The C_(max) of Berberine,Coptisine,Palmatine and Jatrorrhizine were(412.68±28.45),(68.65±3.92),(34.06±3.13),(40.60±1.22)ng/mL,and AUC_(0-∞)were(672.70±72.55),(146.04±25.01),(71.49±18.67),(72.25±9.54)ng·mL^(-1)·h^(-1),respectively.Compared with Jiaotai pill suspension,the Cmax,AUC_(0-t) and AUC_(0-∞)of the four components in insomnia rats were significantly increased(P<0.01).Conclusion:Jiaotai pill self-microemulsionl can promote the absorption of effective components in insomnia rats and improve its bioavailability.

Effect of Jiaotai Pill(交泰丸)on Intestinal Damage in Partially Sleep Deprived Rats

Objective: To explore the effect and mechanism of Jiaotai Pill(交泰丸, JTW) on intestinal mucosal damage in rats with chronic partial sleep deprivation(PSD). Methods: Obesity resistant(OR) rats were selected, and underwent 4 h PSD by being exposed to environmental noise for 4 weeks. During the whole PSD period, JTW and estazolam were orally given to the rats respectively in the treating groups. Plasma concentration of lipopolysaccharide(LPS) which is the marker of gut-origin endotoxemia was examined. Intestinal morphology changes were observed by optical microscopy. The protein expression of occludin(Ocln) in the intestine was measured by immunofluorescence technique and Western blot. The expressions of circadian proteins cryptochromes(Cry1 and Cry2) in the intestine were also determined. Results: The treatment of JTW significantly decreased LPS level in OR rats with PSD(P<0.05). JTW also attenuated insomnia-induced intestinal injury like shorter, sparse and incomplete villus, wide gap between the villus, mucosal swelling and congesting(P<0.05). These changes were associated with the effect of JTW on up-regulating the expressions of Cry1 protein, Cry2 protein and Ocln protein in the intestine. Conclusions: JTW has the beneficial effect on improving intestinal mucosal damage caused by PSD. The mechanism appears to be related to the modulation of the expressions of circadian proteins and Ocln protein in the intestine, thereby attenuating inflammation and improving insulin resistance in insomnia rats.

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.

Effects of Berberine and Cinnamic Acid on Palmitic Acid-Induced Intracellular Triglyceride Accumulation in NIT-1 Pancreatic β Cells

Objective： To investigate the effects of berberine （BBR） and cinnamic acid （CA）, the main active components in Jiaotai Pill （交泰丸, JTP）, on palmitic acid （PA）-induced intracellular tdglyceride （TG） accumulation in NIT-1 pancreatic 13 cells. Methods： Cells were incubated in culture medium containing PA （0.25 mmol/L） for 24 h. Then treatments with BBR （10 μmol/L）, CA （100 μmol/L） and the combination of BBR and CA （BBR＋CA） were performed respectively. Intracellular lipid accumulation was assessed by Oil Red O staining and TG content was measured by colorimetric assay. The expression of adenosine monophosphate-activated protein kinase （AMPK） protein and its downstream lipogenic and fatty acid oxidation genes, including fatty acid synthase （FAS）, acetyl-coA carboxylase （ACC）, phosphorylation acetyl-coA carboxylase （pACC）, carnitine acyl transferase 1 （CPT-1） and sterol regulating element binding protein lc （SREBP-lc） were determined by Western blot or real time polymerase chain reaction. Results： PA induced an obvious lipid accumulation and a significant increase in intracellular TG content in NIT-1 cells. PA also induced a remarkable decrease in AMPK protein expression and its downstream targets such as pACC and CPT-I. Meanwhile, AMPK downstream lipogenic genes including SREBP-lc mRNA, FAS and ACC protein expressions were increased. Treatments with BBR and BBR＋CA, superior to CA, significantly reversed the above genes changes in NIT-1 pancreatic 13 cells. However, the synergistic effect of BBR and CA on intracellular TG content was not observed in the present study. Conclusion： It can be concluded that in vitro, BBR and BBR＋CA could inhibit PA-induced lipid accumulation by decreasing lipoqenesis and increasin.cl lipid oxidation in NIT-1 pancreatic B cells.