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This study aimed to investigate the mechanism of Jiu Wei Bu Xue Oral Liquid on insomnia rats combining the methods of network pharmacology, molecular docking and experimental verification. UPLC-Q-TOF-MS/MS method and TCMIP, TCMSP databases were used to collect the ingredients and targets of Jiu Wei Bu Xue Oral Liquid. Protein-protein interactions and network analysis were performed to screen the key network targets and putative active ingredients of Jiu Wei Bu Xue Oral Liquid in treatment of insomnia, and then following by biological function and KEGG pathway analysis. Then binding ability for key network targets and putative active ingredients were predicted with molecular docking. The prediction targets were validated in para-chlorophenylalanine (PCPA) induced insomnia rats with administration of Jiu Wei Bu Xue Oral Liquid (2, 4, 8 mL·kg-1) for 7 days. Pentobarbital sodium induced sleeping test were performed to evaluate the synergistic sleep-aiding effect of Jiu Wei Bu Xue Oral Liquid. Then glutamic acid (Glu), γ-aminobutyrate (GABA) content and glutamate decarboxylase 1 (GAD67) activity in hypothalamus or hippocampus were evaluated, and the expressions of GAD67, γ-aminobutyric acid receptor subunit α1 (GABRA1) and γ-aminobutyric acid receptor subunit β2 (GABRB2) in hippocampus were detected by qRT-PCR and Western blot methods. Animal experiments were approved by the Institutional Committee on Animal Care of Guangxi Institute of Chinese Medicine & Pharmaceutical Science (the number of permission: 2022060802). Results showed that 16 key network targets and 16 putative active ingredients were obtained by analyzing the herbs-ingredients-targets network of Jiu Wei Bu Xue Oral Liquid in treatment of insomnia. Network pharmacology and molecular docking all indicated these active ingredients, for example atractylenolide Ⅲ, showed better binding ability with GABRA1 and GABRB2. Animal study indicated that, compared to PCPA-induced insomnia model, Jiu Wei Bu Xue Oral Liquid remarkably shortened the sleeping latency and increased the sleeping duration, increased GAD67 activity and the production of GABA in hippocampus of insomnia rats, as well as the expressions of GAD67, GABRA1 and GABRB2, while decreased Glu content in hypothalamus, leading to decreasing of Glu/GABA ratio and recovery of Glu-GABA balance. These results indicated that Jiu Wei Bu Xue Oral Liquid improved insomnia symptoms and helped maintain the Glu-GABA balance within hypothalamus and hippocampus, and reduced the excitatory neurotoxicity within brain. The mechanism may due to the elevation of GAD67 expression and enzyme activity, and the enhancement of type-A GABA receptor (GABAAR)-mediated neurons inhibition.
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Objective: To optimize the extraction technology of Guizhi Zhumian Capsules (GZC). Methods: Based on the analysis of the single factor experiment Results:, the orthogonal test method was adopted to study the three factors including the amount of water added, extraction time, and extraction frequency by taking the dry extract yield of the medicinal materials and transfer rate of geniposide as indicators to optimize the water extraction process of GZC. To optimize the alcohol precipitation process of GSC, these factors including the concentration of the medicinal materials, alcohol content, and time of alcohol precipitation were investigated. The extracts before and after alcohol precipitation were compared by improving sleep pharmacodynamics. Results: The best water extraction technology of GZC was decocted three times with 10 times of water, 0.5 h each time. The optimal alcohol precipitation process was to concentrate the filtrate of water extraction to 1 mL, which was equivalent to 1 g of the original medicinal materials, with 80% alcohol content and 12 h alcohol precipitation time. Compared with the negative control group, the water extract group of GZC could prolong the sleep time of pentobarbital sodium mice and increase the sleep rate of mice under the lower dose of pentobarbital sodium valve (P 0.05). Conclusion: The pharmaceutical efficacy of the extract before and after alcohol precipitation was different. In order not to affect the drug effect, the water extraction process was finally selected as the best extraction process.
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OBJECTIVE: To compare the effects of different anesthetics and blood sampling methods on blood routine test results in experimental animals. METHODS: A total of 42 specific pathogen free( SPF) male Sprague Dawley( SD) rats and 59 SPF male Kunming( KM) mice were randomly divided into 4 groups( control group,ether group,chloral hydrate group and pentobarbital sodium group). Ether group animals were treated with ether inhalation anesthesia; animals in chloral hydrate group and pentobarbital sodium group were injected intraperitoneally with chloral hydrate or pentobarbital sodium. The control group received no anesthesia treatment. Blood samples were collected by different ways: orbital venous plexus,abdominal aorta or eyeball enucleation. White blood cell( WBC) count,red blood cell( RBC) count,platelet(PLT) count,hemoglobin(Hb) level and hematocrit(HCT) in blood samples were analyzed. RESULTS: The RBC count,Hb level and HCT of SD rats in pentobarbital sodium group were significantly lower than those in control group( P <0. 05). The HCT of SD rats in ether group was lower than that in control group( P < 0. 05). The WBC count of orbital venous plexus of KM mice was lower than that taken by eyeball enucleation in control group( P < 0. 05),but the WBC count of orbital venous plexus was higher than that taken by eyeball enucleation in chloral hydrate group( P < 0. 05). The RBC count,Hb level,HCT of KM mice in pentobarbital sodium group were significantly lower than those in control group(P < 0. 05). CONCLUSION: The anesthetic can affect the blood routine test results of experimental animals. Different blood sampling methods have effects on blood routine test results of KM mice.
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Objective To investigate the effectiveness of three different anesthetic techniques in intraventricular catheterization and its effect on the survival rate of rats. Methods Thirty Wistar rats were equally allocated into 3 groups:chloral hydrate group,pentobarbital sodium group and isoflurane group. Intraventricular catheterization was performed in the rats after anesthesia with i. p. injection of chloral hydrate and pentobarbital sodium, and isoflurane inhalation, respectively. Levels of blood glucose were detected before and at 15 and 30 minutes and 1, 3, 7, 14, 28 days after anesthesia. Body mass and 24-hour food intake were recorded before and at 1, 3, 7 days after anesthesia. The onset time and effective time of anesthesia, operation time and the survival rates on 30 days of the rats were compared and analyzed. Results The onset time and effective time of anesthesia, and the operation time in the isoflurane group were shorter than that in the chloral hydrate group, while these parameters in this group were shorter than that in the pentobarbital sodium group. Blood glucose in the chloral hydrate group was apparently increased during the surgical operation, while the body mass, 24-hour food intake and blood glucose were decreasing since one day after operation, and all the rats in this group died during the 30-day observation, mainly, due to enteroplegia. Blood glucose in the pentobarbital sodium group was mildly increased after anesthesia, while the body mass, 24-hour food intake and blood glucose were mildly decreased at one day after operation and recovered within one week. In this group, 3 rats died of respiratory distress due to overdose anesthesia and one rat died during the 30 day-observation. The blood glucose in the isoflurane group was mildly increased after operation, while the 24-hour food intake and blood glucose did not markedly changed, the body mass was stably increased, and no rat died during the 30-day-observation. Conclusions Intraperitoneal injection of chloral hydrate is not suitable for intraventricular catheterization in rats. Intraperitoneal injection of pentobarbital sodium can be only carefully applied for intraventricular catheterization under poorly-limited conditions. Isoflurane inhalation anesthesia is recommended for intraventricular catheterization in rats.
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Objective To compare the effects of isoflurane and pentobarbital on the establishment of subarachnoid block model in rats.Methods 60 SD rats aged 4 months were randomly divided into Group A (n =30) and Group B (n=30).Rats in Group A received intraperitoncal injection of 10 g/L pentobarbital sodium solution 30 mg/kg and 1/4 of the initial dosage was added according to the operation effect.The induction and maintenance of anesthesia were achieved by isoflurane inhalation in Group B during operation.We recorded the time of anesthesia induction,quality of anesthesia,time of anesthesia,time of operation,and recovery time.The heart rate,respiration frequency,temperature,and saturation of blood oxygen were recorded during operation.We compared death from anesthesia and success of modeling in the two groups.Results There was no significant difference between the groups with regard to age,weight,body temperature or saturation of blood oxygen (P＞ 0.05).Compared to Group B,heart rate decreased 1-60 minutes after anesthesia and respiration frequency decreased 5 minutes after anesthesia in Group A (P＜0.05).The time of anesthesia induction,time of anesthesia,time of operation,and recovery time were shorter in Group B (P＜0.05).The quality of anesthesia was better in Group B (P＜0.05).The success rate of modeling was higher but mortality rate of anesthesia was lower in Group B than in Group A (P＜0.05).Conclusion Compared with intraperitoneal injection of pentobarbital sodium,isoflurane inhalation can provide a better anesthetic effect during the operation to establish a rat model of subarachnoid block.
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Aim To study the hypnotic effect of five alkaloids extracted from Rhizoma Coptidis (berberine , coptisine,palmatine,epiberberine,jatrorrhizine )in mice,and preliminarily explore its underlying mecha-nism.Methods The experiments of locomotor activity and hypnosis induced by suprathreshold and subthresh-old doses of pentobarbital sodium were used to evaluate the effect of drugs on sleep behavior in mice.Then, HPLC-FLD was used to detect the contents of NE,DA and 5-HT on PCPA mice model.Results Compared with control group,berberine and coptisine notably in-hibited spontaneous activity in behavioral experiments (P <0.05),and increased the sleeping percentage of mice under subthreshold dose of pentobarbital sodium. Berberine and coptisine shortened the period of sleep latency,and prolonged the sustained period of sleeping at suprathreshold dose in mice (P <0.05 or P <0.01 ).Other alkaloids had no significant differences in sleep latency and period of sleep observed in this current experiment.Compared with PCPA mice model group,berberine and coptisine remarkably increased the contents of NE and 5-HT (P <0.01 ),but they had no effects on DA.Conclusions Berberine and coptisine may play a sedative and hypnotic role in PC-PA mice by increasing contents of 5-HT and NE in hy-pothalamus,and the sedative and hypnotic effects of berberine are stronger than those of coptisine.Other alkaloids have no effects on sleeping in mice.
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Objective To observe the effects of mild hypothermia induced by pentobarbital sodium on hematology in male BALB/C mice.Method Healthy male BALB/C mice were divided randomly into two groups:control group ( C) and mild hypothermia group(M).The body temperature of the mild hypothermia group was maintained between 28℃ to 30℃( anal temperature ) for 4 hours induced by pentobarbital sodium injected intraperitoneally , then recover unaffected . Anal temperature, coagulation, electrolytes, and blood cell indexes were examined in 2, 24, 72 hours after treated by mild hypothermia;Control group was given equal volume of saline volume at constant temperature .Results The body temperature and coagulation in mild hypothermia group showed no significant difference compared with the control group ( P﹥0.05),but the concentration of K +and Na +in mild hypothermia group were higher than control group (P﹤0.01), the number of WBC in mild hypothermia group was lower than control group ( P﹤0.01或P﹤0.05 ) , and the RBC、HGB、MCH、MCHC in mild hypothermia group were lower than control group transiently (P﹤0.01或P﹤0.05).Conclusion Mild hypothermia induced by pentobarbital sodium affects some of hematological values in mice considerably .
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Aim To establish a novel locomotor model based on accumulate peripheral active time percent(APATP)in ICR mice induced by pentobarbital sodium(PB)and diazepam.Methods Total distance, accumulate peripheral active time and accumulate peripheral time were aquired from video tracking system with computer, and APATP in all groups was fit and got correspondent parameters with Drugs Analysis System 2.0. Logarithm of area under the curve(lgAUC_(0-60)), minimum APATP(lgP_(min)), and half time of steady-state locomotor activity(T_(1/2α)) were ready for evaluation.Results APATP decreased with time increasing in all groups gradually, and PB 10 mg·kg~(-1) and PB 15 mg·kg~(-1) had similar tendency.PB 10 mg·kg~(-1) and PB 15 mg·kg~(-1) all decreased APATP significantly throughout the time course.APATP in all mice was fit to kinetics equation.Compared with norm group, PB 5 mg·kg~(-1) decreased three parameters slightly(all P >0.05), and PB 10 mg·kg~(-1) and PB 15 mg·kg~(-1) had similar tendency.PB 10 mg·kg~(-1) (all P <0.05) and PB 15 mg·kg~(-1) (P <0.01, P <0.01 and P >0.05)decreased parameters.Compared with PB 10 mg·kg~(-1) , PB 15 mg·kg~(-1) increased reversely(P >0.05, P <0.05 and P <0.05).lgAUC_(0-60) and lgPmin were both linar with total distance(r 2=1.0000 and r 2=0.9995, both P <0.01), T_(1/2α) also showed similar tendcency as well as total distance.Refering PB 10 mg·kg~(-1) as positive drugs and norm as negative control, diazepam 2 mg·kg~(-1) and 4 mg·kg~(-1) depressed all parameters significantly compared with norm group(all P <0.01) and were similar as PB 10 mg·kg~(-1) , which indicated that sedative effect of diazepam was the same as PB 10 mg·kg~(-1) .Conclusion Locomotor activity model based on APATP may be used to evaluate drug effects on lomocotor activity induced by sedative hypnotics.
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Objective To observe the influence of the coumarins from Radix Angelicae pubescentis (TCA) on the hypnotic effects of pentobarbital sodium and barbital sodium. Methods Mice were randomly divided into solvent (5% Tween-80, 20 mL/kg), chloromycetin (25 mg/kg) and TCA (50, 100 mg/kg) groups, which were administered orally with equivalent volume. One hour later, all the animals were given pentobarbital sodium (40 mg/kg, ip) or barbital sodium (200 mg/kg, ip), and then the latent period and duration of hypnosis were observed. Results Hypnotic duration of pentobarbital sodium was prolonged by 249.8%, 171.1% and 78.7% in chloromycetin and two TCA groups respectively (P
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Objective To investigate the preventive and therapeutic effects of Ginger extraction on heart failure in rabbits.Methods Twenty-seven New Zealand pure-breed rabbits were randomized into 3 groups:group A,group B and group C.Group A was orally administrated with 2 %Ginger extraction 3 days before and after modeling;group B was orally administrated with purified water 3 days before modeling and with 2 %Ginger extraction after modeling;group C was orally administrated with purified water 3 days before and after the establishment.The differences of modeling time and dosage of pentobarbital sodium between groups were compared;hemodynamics parameters before and after administration were detected.Results (1)The modeling time and dosage of Pentobarbital Sodium were remarkably increased in group A,differences being significant compared other 2 groups(P