Synergistic action on hypnosia: Yinao capsules with pentobarbital sodium of threshold and sub-threshold dosages

BACKGROUND： Sedative and hypnotic drugs could cure insomnia in a dependent manner, and traditional Chinese medicine has some superiority in treating insomnia. OBJECTIVE： To observe the synergistic action of yinao capsules with pentobarbital sodium in threshold and sub-threshold dosages in hypnosia and sedation. DESIGN： A completely randomized grouping design and control experiment. SETTING： Pharmacological laboratory, College of Pharmacy, Henan College of Traditional Chinese Medicine. MATERIALS： Totally 200 grade II Kunming mice of 18 - 21 g, either male or female, were used. Yinao capsules, main ingredients of which were turtleback glue, thinleaf milkwort root, Chinese magnoliavine fruit, mythic fungus, tangshen, ginseng and grassleaf sweetflag rhizome, were offered by Chinese-American Huayi Pharmacy, Co.,Ltd. （ratified number： 040901）; Kangnaoshuai capsules, main ingredients of which were prepared rehmannia root, tuber fleeceflower root, ginseng, membranous milkvetch root, thinleaf milkwort root, Fushen, grassleaf sweetflag rhizome, spine date seed, lecithin, barbary wolfberry fruit, pueraria root, vitamin E, etc., were produced by Shijiazhuang Siyao, Co.,Ltd. （ratified number： 040964）; Pentobarbital sodium was produced by China Medicine （Group） Shanghai Chemical Reagent, Co,.Ltd. （Ratified number： 030816）. ZZ-6 mice spontaneous activity apparatus was produced by Chengdu Taimeng Science and Technology, Co.,Ltd. METHODS： The experiment was carried out in the Animal Experimental Center, Henan College of Traditional Chinese Medicine from October to December in 2005. （1） Influence of Yinao capsules on the spontaneous activity of mice： Fifty mice were randomly divided into five groups with 10 mice in each group： Mice in the large, middle and small dosages of Yinao capsules groups were intragastrically infused with Yinao capsules suspension （1.36, 0.68, 0.34 g/kg）; Those in the Kangnaoshuai capsules group were infused with Kangnaoshuai suspension （1.12 g/kg）; Those in the control group were given physiologic saline of the same volume. The mice were administrated once a day for 7 days continuously, and they were placed into the mice spontaneous activity apparatus after 60 minutes from the last administration, the times of spontaneous activities and the times of arising within 10 minutes were recorded after adaptation for 5 minutes. （2） Synergistic action in hypnosia by Finao capsules with pentobarbital sodium of threshold dosage： Seventy mice were randomly divided into 5 groups as above-mentioned with 14 mice in each group, and they were treated the same as above. They were intraperitoneally injected with 50 mg/kg pentobarbital sodium after 60 minutes from the last administration, then conditions of falling asleep were observed. The disappearance of righting reflex was taken as the index of falling asleep, the latency of falling asleep was the duration from intraperitoneal injection of pentobarbital sodium to fall asleep, the sleeping time was from falling asleep to the disappearance of righting reflex. （3） Synergistic action in hypnosia by Yinao capsules with pentobarbital sodium of sub-threshold dosage： Eighty mice were randomly divided into 5 groups as above-mentioned with 16 mice in each group, and they were treated the same as above. They were intraperitoneally injected with 27 mg/kg pentobarbital sodium after 60 minutes from the last administration. Mice whose righting reflex disappeared for at least 1 minute within 30 minutes were taken as falling asleep, the number of sleeping animals in each group was recorded, and the rate of falling asleep was calculated. MAIN OUTCOME MEASURES： Synergistic action of Yinao capsules with pentobarbital sodium of threshold and sub-threshold dosages in hypnosia and sedation. RESULTS： All the 200 mice were involved in the analysis of results. （1） Influence of Yinao capsules on the spontaneous activity of mice： The times of spontaneous activities within 10 minutes in the large and middle dosage of Yinao capsules groups and Kangnaoshuai capsules group [（138.0±37.0）, （156.8±28.3）, （133.3±46.1） times] were obviously fewer than those in the control group [（204.3±61.3） times, P 〈 0.05- 0.01]. The arising times within 10 minutes in the middle and small dosages of Yinao capsules groups and Kangnaoshuai capsules group [（30.7 ± 18.3）, （26.5± 11.2）, （24.2±11.6） times] were obviously fewer than those in the control [（71.7±38.6） times, P 〈 0.01 ]. （2） Synergistic action in hypnosia by Yinao capsules with pentobarbital sodium of threshold dosage： The sleeping latencies in the large and middle dosages of Yinao capsules groups and Kangnaoshuai capsules group [（4.49±1.84）, （4.83±1.72）, （3.85± 1.94） minutes] were obviously shorter than that in the control group [（6.73 ±2.75） minutes, P 〈 0.05 - 0.01 ]. The sleeping time in the large, middle and small dosages of Yinao capsules groups and Kangnaoshuai capsules group [（89.0± 38.42）, （67.21 ±24.07）, （66.28±18.94）, （84.36±29.81） minutes] were obviously longer than that in the control group [（45.78±20.78） minutes, P 〈 0.05- 0.01]. （3） Synergistic action in hypnosia by Yinao capsules with pentobarbital sodium of sub-threshold dosage： The rates of falling asleep in the large and middle dosages of Yinao capsules groups were higher than that in the control group [56% （9/1 6）, 38% （6/1 6）, 6% （6/16）, P 〈 0.01, 0.05], whereas Kangnaoshuai capsules and small dosage of Yinao capsules had no synergistic action with pentobarbital sodium of sub-threshold dosage （P 〉 0.05）. CONCLUSION： Yinao capsules have synergistic action with pentobarbital sodium in hypnosia. Yinao capsules possess obvious sedative and hypnotic effects in a dosage-dependent manner.

Embryonic skeleton development and neonatal learning and memory ability of rats anesthetized with pentobarbital sodium: Differences of administration occasion and time

BACKGROUND: Generally speaking, anesthesia is often used in gravid body and it has been already proved that many kind of medicine can result in malformation. OBJECTIVE: To explore embryonic skeleton development and neonatal learning and memory of rats anesthetized with pentobarbital sodium in gravid rats. DESIGN: A randomized control trial. SETTING: Laboratory Animal Center of Xuzhou Medical College. MATERIALS: A total of 80 adult female SD rats, of clean grade and weighing 220-240 g, were selected in this study. The main reagents were detailed as follows: pentobarbital sodium (Shanghai Xingzhi Chemical Plant, batch number: 921019); MG-2 maze test apparatus (Zhangjiagang Biomedical Instrument Factory); somatotype microscope (Beijing Taike Instrument Co., Ltd.). METHODS: ① A total of 160 SD rats of half males and females were selected in this study. All rats were copulated. The day that the plug was checked out in the vagina next day was looked as the first day of pregnancy. Gravid rats were divided randomly into four groups, including early anesthesia group, second anesthesia group, late anesthesia group and control group with 20 in each group. Rats in the early anesthesia group were injected with 25 mg/kg soluble pentobarbitone on the 7th day of pregnancy for once; rats in the second anesthesia group were anesthetized with 25 mg/kg soluble pentobarbitone on the 7th and the 14th days of pregnancy for once; rats in the late anesthesia group were anesthetized with 25 mg/kg soluble pentobarbitone on the 14th day of pregnancy for once; rats in the control group did not treat with anything. The time of anesthetizing was controlled in 3 to 4 hours and ether was absorbed while the time was not enough. ② Half of each group was sacrificed on day 20th of pregnancy and the fetus was taken out to be stained with alizarin red S. After stained, the fetal skeleton was examined. The learning and memorizing of one-month rats that were given birth by the rest gravid rats were tested through electric mare method. Determine their study ability according to their correct rate of 90% or above of arrival at the safe area in 20 s. After they finally learned to arrive at the safe area correctly, test them once more in 24 hours and record the correct rate of 15 times. MAIN OUTCOME MEASURES: The rate of malformation in fetus and ability of learning and memory in one-month rats. RESULTS: A total of 80 female rats were anesthetized in this experiment. Totally 490 immature rats were tested with maze testing machine and 196 fetuses were stained with alizarin red S to observe the development of their skeleton. However, one of the 80 female rats was led to death because of overdose. ① Malformation experiment: Learning ability of second anesthesia group was evidently different from the control group while the other two groups were not in the electric mare method. The fetal skeleton malformation rate of three experimental groups was 87.0%, 60.9% and 17.9%, respectively, while it was 5.6% in the control group. ② Electric mare method: Times of rats which arrived at the safe regions were respectively 49.0±31.0, 68.0±35.0, 47.0±31.0 and 44.0±21.0 in early anesthesia group, second anesthesia group, late anesthesia group and control group; and then, there was significant difference between the second anesthesia group and the control group (P < 0.05). Exact rates of memory of rats were respectively (64.36±14.35)%, (62.15±18.33)%, (54.19±12.28)% and (68.24±15.91)% in early anesthesia group, second anesthesia group, late anesthesia group and control group; and then, there were no significant differences as compared with the control group (P > 0.05). CONCLUSION: The influence of anesthesia with pentobarbital sodium is obvious in fetal skeleton development and learning and memory ability.

Brain interstitial fluid drainage and extracellular space affected by inhalational isoflurane: in comparison with intravenous sedative dexmedetomidine and pentobarbital sodium

Brain interstitial fluid drainage and extracellular space are closely related to waste clearance from the brain. Different anesthetics may cause different changes of brain interstitial fluid drainage and extracellular space but these still remain unknown. Herein,effects of the inhalational isoflurane, intravenous sedative dexmedetomidine and pentobarbital sodium on deep brain matters’ interstitial fluid drainage and extracellular space and underlying mechanisms were investigated. When compared to intravenous anesthetic dexmedetomidine or pentobarbital sodium, inhalational isoflurane induced a restricted diffusion of extracellular space, a decreased extracellular space volume fraction, and an increased norepinephrine level in the caudate nucleus or thalamus with the slowdown of brain interstitial fluid drainage. A local administration of norepinephrine receptor antagonists, propranolol,atipamezole and prazosin into extracellular space increased diffusion of extracellular space and interstitial fluid drainage whilst norepinephrine decreased diffusion of extracellular space and interstitial fluid drainage. These findings suggested that restricted diffusion in brain extracellular space can cause slowdown of interstitial fluid drainage, which may contribute to the neurotoxicity following the waste accumulation in extracellular space under inhaled anesthesia per se.

Effect of different anesthesia methods on erythrocyte immune function in mice

Objective:To explore effect of different anesthesia methods and different anesthetics on erythrocyte immune function in mice.Methods:The mice were anesthetized by isoflurane and ether inhalation,and also under intraperitoneal anesthesia with sodium pentobarbital and chloral hydrate,Blood was collected from the ventro-cardinal vein.Automatic blood cell analyzer was used for routine blood examination,and the canthine oxidase method was used to measure the superoxide dismutase(SOD)activity.Lipid peroxidation product malondialdehyde(MDA)was measured with TBA,and glutathione peroxidase(GSH-Px)was measured with DTNB,and then the effect of different anesthesia methods and different anesthetics on erythrocyte immune function in mice was observed.Result:Hct level of chloral hydrate intraperitoneal injection group was significantly higher than the other three groups(P<0.05).And the MDA levels in the pentobarbital sodium group were significantly higher than the other three groups(P<0.05).SOD and GSH-Px of the chloral hydrate and sodium pentobarbital intraperitoneal injection group were significantly lower than the other two groups;RBC-C 3bRR and RBC-ICR of the chloral hydrate and sodium pentobarbital intraperitoneal injection group were significantly lower than the other two groups.Conclusions:Different drugs can induce changes in immune function of mice at different levels.Isoflurane and ether have less damage to animal body,while chloral hydrate and sodium pentobarbital intraperitoneal injection have a certain inhibitory effect on the animal body respiratory system and can cause greater damage to the body.Therefore,the reasonable selection and control of anesthetics are very important in order to avoid the experimental errors caused by anesthesia.

Anshen Babu Ointment Improves Sleep Function of Insomnia Mice and Its Mechanism

[Objectives]This study aimed to study the sleep-improving effect of Anshen Babu ointment on insomnia mice and its mechanism of action.[Methods]Mouse models of insomnia were established by p-chlorophenylalanine(PCPA)injection.They were given high,middle and low doses of Anshen Babu ointment,for 7 consecutive days,and then,the spontaneous activity of insomnia mice and the effect on hypnotic time and subthreshold hypnotic dose of pentobarbital sodium were observed,and the levels of 5-HT,IL-6 and TNF-α in the hippocampus of mice were determined.[Results]High and middle-dose Anshen Babu ointment significantly reduced the spontaneous activity of insomnia mice and shorten the hypnotic time of pentobarbital sodium;high and middle-dose Anshen Babu ointment increased the 5-HT content and inhibited the IL-6 and TNF-α expression in the hippocampus of mice.[Conclusions]Anshen Babu ointment has hypnotic and sedative effect on insomnia mice.The mechanism may be achieved by increasing 5-HT and reducing IL-6 and TNF-α levels.