臭椿生物碱canthin-6-one对小鼠镇静催眠作用的研究

摘要 目的：研究臭椿生物碱canthin-6-one对小鼠的镇静催眠作用。方法：选昆明小鼠随机分为空白组、地西泮阳性药物组、canthin-6-one低、中、高给药组（5、10、20 mg?kg^-1），采用阈上剂量和阈下剂量戊巴比妥钠诱导小鼠睡眠，记录入睡小鼠比例、睡眠潜伏期和睡眠持续时间。此外，酶联免疫法测定小鼠脑组织中5-HT、NE、DA和GABA含量。结果：中、高剂量canthin-6-one可使小鼠的站立次数和自主活动次数均明显减少（P＜0.05），可使阈上剂量戊巴比妥钠诱导的小鼠的睡眠潜伏期缩短（P＜0.05）；各剂量组可使阈下剂量戊巴比妥钠诱导的小鼠入睡率提高（P＜0.05）。此外，中、高剂量canthin-6-one也可降低小鼠脑组织中5-HT和NE含量，增加GABA含量（P＜0.05），对DA的含量有影响，但无统计学意义（P＞0.05）。结论：Canthin-6-one有明显的协同戊巴比妥钠改善睡眠作用，其作用与相关神经递质（5-HT、NE、GABA）具有密切关系。     

天麻改善小鼠睡眠作用研究

目的:探讨天麻粉对小鼠改善睡眠作用的影响。方法:144只雌性KM小鼠随机分为三个实验组,每个实验组48只,分别设置低、中、高三个剂量组,分别给予0.4、0.8、1.2 g/kg BW天麻粉,同时设置阴性对照组(给予等量蒸馏水),各组均为12只(n=12)。给予小鼠连续灌胃30天后开始实验。实验一组进行直接睡眠实验和戊巴比妥钠阈下剂量催眠实验,实验二组进行巴比妥钠睡眠潜伏期实验,实验三组进行延长戊巴比妥钠睡眠时间实验。结果:与阴性对照组比较,天麻粉各剂量组小鼠终重均没有明显改变(p0.05);高剂量组提高阈下剂量戊巴比妥钠诱导的小鼠睡眠发生率作用,差异有统计学意义(p0.05);中、高剂量组明显缩短巴比妥钠睡眠潜伏期时间,差异有显著性统计学意义(p0.01);高剂量组明显延长戊巴比妥钠诱导小鼠睡眠时间的作用,差异有显著统计学意义(p0.01)。结论:天麻粉对小鼠具有一定的改善睡眠作用,且对小鼠体重增长无影响。     

人参对睡眠促进作用的实验研究

目的研究人参对睡眠促进作用。方法选择ICR小鼠,随机分为空白对照组和阳性对照组。A组(50%醇提液)、B组(75%醇提液)、C组(90%醇提液)、D组(水提液)、阳性对照组6组。A-D组为实验动物组,经灌胃途径按照50g/kg剂量给药,空白对照和阳性对照组给予相同体积蒸馏水,连续灌胃给药7d,末次给药30min后,阳性对照组经腹腔注射地西泮2.5mg/kg,其余经腹腔注射给予50mg/kg的戊巴比妥钠,空白对照和阳性对照组给予相同体积蒸馏水,记录睡眠潜伏时间、睡眠持续时间。结果阳性对照组比空白对照组能缩短小鼠睡眠潜伏期且可以延长睡眠时间(p0.05)。与空白对照组比,A组(50%醇提液)和B组(75%醇提液)均有明显差异,B组明显缩短了睡眠潜伏期和延长睡眠时间,差异具有显著性意义(p0.05)。结论75%醇提的人参改善睡眠作用效果最佳,为后续开发具有改善睡眠功能和记忆功能的中药复方制剂提供了科学依据。     

灵芝酸枣仁胶囊改善睡眠的功能研究

为研究灵芝酸枣仁胶囊的改善睡眠功能,将ICR小鼠随机分为灵芝酸枣仁高、中、低剂量组和空白对照组,每组12只。人体推荐每公斤服用剂量为45 mg/kg·d,以此剂量的5倍、10倍和30倍,即每日以225 mg/kg BW、450 mg/kg BW和1 350 mg/kg BW经口灌胃30 d后,通过直接睡眠、延长戊巴比妥钠睡眠时间、巴比妥钠阈下剂量催眠和巴比妥钠睡眠潜伏期实验评价灵芝酸枣仁胶囊的改善睡眠作用。研究发现,与空白对照组相比,3个剂量组的直接睡眠实验结果皆为阴性(P0.05);与空白对照组相比,中剂量组小鼠延长戊巴比妥钠睡眠时间显著延长(P0.05),而高剂量组小鼠戊巴比妥钠睡眠时间极显著延长(P0.01);与空白对照组相比,高剂量组小鼠的入睡只数具有显著差异(P0.05),而睡眠潜伏期的小鼠只数具有极显著差异(P0.01)。研究表明,灵芝酸枣仁胶囊具有改善睡眠作用。     

化学药物对戊巴比妥钠生物转化的刺激和抑制

以小鼠戊巴比妥钠睡眠时间,戊巴比妥钠在小鼠体内消失速率及大鼠肝切片对戊巴比妥钠的代谢为指标观察了几种药物对戊巴比妥钠转化的影响。按对戊巴比妥钠睡眠时间的影响,可将这些药物分为三类:甲类包括密尔通,苯妥英钠,苯海拉明,氨基比林,氯丁醇,氯丙嗪及3′-甲基奶油黄等已知的药物转化酶刺激剂,给这些药物后48小时,小鼠戊巴比妥钠睡眠时间缩短,但在给药后1小时睡眠时间延长。由于给这些药后1小时戊巴比妥钠在体内的消失延缓,以及当温孵液中含有6.6—17.0×10~(-4)M 时明显抑制大鼠肝切片对戊巴比妥钠的代谢,故这些药物延长睡眠时间的原因,至少部分由于抑制催眠药的生物转化。乙类药物只延长睡眠时间而不随后使之缩短,包括丙嗪,美沙酮,E605,安他布斯,PT-22,2-甲基奶油黄,奶油黄,牛胱胺,AET 及氮芥类化合物。其中美沙酮,E605,安他布斯及2-甲基奶油黄都经进一步证明能抑制戊巴比妥的转化,新恩比兴在给药后第3天有抑制作用,唯丙嗪并不延缓戊巴比妥钠在小鼠体内的消失。丙类包括阿司匹林,DFP,6-MP 及 8-氮杂鸟扁便嘌呤等,既不在给药后早期显著延长睡眠时间,也不在后期缩短睡眠时间。上述结果表明:凡药酶刺激剂,在给予动物的早期,必表现出对药酶的抑制作用;但药酶抑制剂在给予动物后晚期,对药酶不一定都有刺激作用。     

硫代乙酰胺对戊巴比妥钠睡眠时间和肝脏药物转化的影响

给小鼠皮下注射硫代乙酰胺60毫克/公斤后48小时戊巴比妥钠睡眠时间明显延长;同剂量的硫代乙酰胺给大鼠后在12—72小时内戊巴比妥钠睡眠时间也都明显延长,而以48小时者最为明显。进一步实验发现48小时前接受一剂硫代乙酰胺处理的小鼠,戊巴比妥钠自体内消失的速率比正常动物者明显减慢;12—72小时前曾接受一剂硫代乙酰胺的大鼠肝切片转化戊巴比妥钠的速率比正常动物肝切片者小,此与戊巴比妥钠睡眠时间的延长相一致。可见,经硫代乙酰胺处理的动物的戊巴比妥钠睡眠时间延长的原因之一,是由于硫代乙酰胺抑制了肝脏对戊巴比妥钠的转化。另一方面,48小时前曾接受过硫代乙酰胺处理的小鼠,睡眠刚醒时,体内戊巴比妥钠含量明显地低于对照组;曾经硫代乙酰胺处理的大鼠刚醒时,脑组织的戊巴比妥钠含量明显低于对照组者。在经硫代乙酰胺处理后的小鼠,戊巴比妥钠引起睡眠的 ED_(50)明显减小;二乙基巴比妥钠的睡眠时间亦明显延长。这些结果说明硫代乙酰胺处理也增加了动物中枢对戊巴比妥类药物的敏感性。     

马齿苋中褪黑素的含量测定及其对小鼠睡眠的影响

目的：建立对马齿苋药材中褪黑素的高效液相色谱检测方法并研究其对小鼠睡眠的影响。方法：以乙醇为溶剂采用超声波法提取褪黑素,利用固相萃取小柱对褪黑素进行富集,安捷伦高效液相色谱仪对褪黑素进行分析检测。将小鼠分为空白、地西泮（1 mg/kg）、褪黑素提取物组（0.5 mg/kg以褪黑素计）,小鼠灌胃给药30 min后,腹腔注射戊巴比妥钠,记录各组小鼠睡眠潜伏期和睡眠时长。结果：褪黑素质量浓度范围在100 ~2 000 ng/mL范围内与峰面积的线性关系良好（R2=0.999 9）,线性回归方程为Y=0.080 5X+0.083 6,平均加样回收率为101.40%,RSD为1.18%。测得马齿苋中褪黑素含量为686.17 ng/g。地西泮组以及褪黑素提取物组小鼠的睡眠潜伏期显著缩短,睡眠时间显著增加。结论：该方法方便快捷,分离效果好,可用于马齿苋中褪黑素含量的测定;马齿苋中所提取的褪黑素可缩减小鼠睡眠潜伏期,提升睡眠时间,具有镇静催眠作用。     

不同品系小鼠对代谢性高尿酸血症造模的影响

目的：分别以昆明种小鼠及ICR、C57BL/6J小鼠为研究对象,比较在复制高尿酸血症模型时可能的小鼠品系差异,并通过降尿酸药物别嘌呤醇与非布索坦验证选择降尿酸药物筛选时选用不同品系动物造模的影响。方法：采用不同剂量次黄嘌呤腹腔注射联用尿酸酶抑制剂氧嗪酸钾皮下注射给药,测定不同造模时段各品系小鼠血清尿酸值。结果：ICR、C57BL/6J小鼠对高尿酸血症造模耐受显著高于昆明种小鼠,在腹腔注射次黄嘌呤500mg/kg,皮下注射氧嗪酸钾300mg/kg时,才可获得稳定的可用于药物筛选的高尿酸血症模型。结论：选择高尿酸血症在体模型时,昆明种小鼠灵敏度高于ICR小鼠以及近交系的C57BL/6J小鼠。     

水迷宫实验中三种品系小鼠学习记忆能力的比较

目的探讨三种品系小鼠在Morris水迷宫实验中的学习记忆能力差异,为与改善学习记忆能力相关新药研究的实验动物选择提供参考。方法选用C57BL/6小鼠、昆明种小鼠和ICR小鼠,进行定位航行实验(5d)、空间搜索实验(1d)和工作记忆实验(4d),考察其学习记忆能力。结果定位航行实验从第3天起C57BL/6小鼠的潜伏期明显小于昆明种小鼠和ICR小鼠(P0.01);空间搜索实验C57BL/6小鼠穿过原平台位置的次数、在原平台象限游程比率和时间比率均明显多于昆明种小鼠和ICR小鼠(P0.05);动物每天工作记忆实验的潜伏期随测试次数增加而缩短,C57BL/6小鼠缩短的较昆明种小鼠和ICR小鼠稍多。结论C57BL/6小鼠的空间参考记忆能力与工作记忆能力均优于昆明种小鼠和ICR小鼠,且非空间因素对其学习记忆能力的评价干扰较小,可作为优先选择的水迷宫实验动物。     

复方安神精油吸入给药的镇静催眠作用及化学成分的GC-MS分析

为研究复方安神精油吸入给药的镇静催眠作用,本实验采用动物自主活动测试和戊巴比妥钠诱导的睡眠潜伏期与睡眠时间实验:GC-MS分析复方安神精油的挥发性化学成分与入脑的主要化学成分;基于网络药理学预测和筛选出治疗失眠症相关靶蛋白与活性成分,旨在为复方安神精油治疗与改善失眠症提供理论依据。研究发现,复方安神精油具有镇静催眠作用,可显著减少动物自主活动,缩短睡眠潜伏期,延长睡眠时间;复方安神精油主要化学成分与入脑主要化学成分中含有多种具有神经中枢镇静催眠作用以及抗焦虑、抗抑郁作用的化学成分;基于网络药理学预测和筛选出25个治疗失眠症相关活性成分作用于39个治疗失眠症相关作用靶点,体现复方安神精油多成分、多靶点发挥镇静催眠作用的特点。     

Marked variation in diazepam sensitivity in Swiss albino mice

Using the righting reflex as the critical level, sleep was measured in Swiss albino mice at a dose of 35 mg/kg diazepam, i.p. Sleep times varied markedly from zero to 120 min with a mean +/- s.d. of 44 +/- 37 (N = 202). The distribution is skewed to the left with a coefficient of skewness of 0.33 +/- 0.17. The sleep times of the two sexes, when analyzed separately, showed similar range, mean and s.d., except that the distribution tended to be more clearly bimodal in males than in females. These animals also exhibited marked variations in their response to either ethanol (4 g/kg) or pentobarbital (45 mg/kg). The diazepam sleep time failed to correlate with the ethanol sleep time. Significant correlation, however, was obtained between diazepam and pentobarbital sleep times. On further analysis with least-squares fit to a straight line, the data yielded a line with a slope of 0.16; thus despite the correlation reaching a significant level, there is no significant difference in the pentobarbital sleep times between mice that have the longest or the shortest diazepam sleep times. By monitoring the plasma and brain levels of diazepam and N-desmethyldiazepam in mice at awakening, it was found that the variations in sleep time cannot be explained by individual differences in drug disposition. The phenomenon is discussed in terms of individual variations in diazepam sensitivity and the possibility of development of tolerance to diazepam almost immediately after diazepam administration.     

酮康唑对小鼠肝脏和睾丸生理功能的影响*

目的: 观察不同剂量的酮康唑(KCZ)对昆明小鼠的肝脏和睾丸生理功能的影响。方法: 将 40 只雄性昆明小鼠随机分为 4 组(n=10):正常组、酮康唑低剂量组(30 mg/kg)、酮康唑中剂量组(50 mg/kg)、酮康唑高剂量组(70 mg/kg)。药物组小鼠按0.1 ml/10 g给药体积每日单次皮下注射酮康唑,酮康唑低、中、高剂量组药物浓度各为3 mg/ml、5 mg/ml、7 mg/ml,正常组注射等量生理盐水,连续 3 周。测定血清谷草转氨酶(AST)、谷丙转氨酶(ALT),以及睾丸组织 γ-谷氨酰胺转肽酶(γ-GT)、乳酸脱氢酶(LDH)和酸性磷酸酶(ACP)活性;HE 染色观察肝脏、睾丸病理组织改变。结果: 与正常组比较,酮康唑组AST、ALT活性显著升高(P＜0.01);γ-GT、ACP、LDH活性显著下降(P＜0.01),且以上指标的改变均随剂量增加而损伤加重。HE 染色显示小鼠肝细胞变性,排列松散,胞质色浅;睾丸曲细精管管腔增大,各级生精细胞、精子数量减少。结论: 酮康唑能导致小鼠肝脏、睾丸组织生理功能损伤与病理组织学改变,升高肝转氨酶水平,降低睾丸特异性酶活性,且损伤程度随剂量的升高而增大。     

Potentiation of pentobarbital hypnosis by Rosa damascena in mice

Rosa damascena has been found to act on central nervous system including brain. It inhibits the reactivity of the hypothalamous and pituitary systems in rat. In traditional medicine hypnotic effect of Rose is also suggested. In the present study hypnotic effect of ethanolic, aqueous and chloroformic extracts of R. damascena was investigated in mice. Hypnotic method was based on potentiation of pentobarbital induced sleeping time by extracts. Three doses of extracts (100, 500 and 1000 mg/kg) were injected i.p. in comparison with diazepam (3mg/kg) as positive control and saline as negative control. After 30 min of injection of extracts, pentobarbital (30mg/kg) was injected and increase in sleeping time by extracts was recorded. The results showed that the ethanolic and aqueous extracts in 500 and 1000 mg/kg doses significantly increased pentobarbital induced sleeping time which was comparable to diazepam. The chloroformic extract had no hypnotic effect.     

The effect of long-term nitrazepam administration on sleep cycles in rats]

Single and chronic administration of a low dose of nitrazepam (1 mg/kg) had no effect on sleep cycles in rats. A single injection of a high dose (10 mg/kg) of nitrazepam resulted in prolongation of the total duration of synchronized sleep with a corresponding shortening of desynchronized (paradoxical) sleep. The number of sleep cycles was reduced. Chronic injections of nitrazepam (for 7-14 days) in a dose of 10 mg/kg evoked a gradual prolongation of the duration of paradoxical sleep and an increase in number of sleep cycles. After discontinuance of a long-term administration of nitrazepam (1 mg/kg or 10 mg/kg) prolongation of desynchronized sleep and an increase in the number of sleep cycles were more pronounced in comparison with the last day of chronic administration of the drug.     

两种发酵食品对糖尿病模型小鼠血糖及血脂的影响

目的：观察诺丽果汁和纳豆两种发酵食品对实验性糖尿病小鼠血糖及血脂的影响。方法：ICR雌性小鼠一次性尾静脉注射四氧嘧啶（55 mg/kg）,72 h后将空腹血糖值≥ 12.00 mmol/L、尿糖呈强阳性（+++）者视为糖尿病小鼠模型。将糖尿病模型小鼠随机分为3组（n=10）：模型（DM）组、诺丽果汁（NJ）组和纳豆（NT）组,另取10只正常ICR雌性小鼠作为正常对照（NC）组。NJ组、NT组分别给予诺丽果汁（25.0 ml/kg）、纳豆（0.6 g/kg）灌胃,其他两组小鼠分别给予生理盐水（25.0 ml/kg）灌胃,连续给药30 d,小鼠自由进食、饮水,记录小鼠饮水量及进食量。末次给药后1.5 h,测定小鼠葡萄糖耐量,经股动脉采血测定小鼠糖化血清蛋白（GSP）、血清胰岛素（Ins）和血脂等指标的变化情况。结果：与NC组比较,DM组小鼠饮水量、进食量、GSP及血清总胆固醇（TC）、甘油三酯（TG）、低密度脂蛋白（LDL）等均显著升高（P<0.01）,葡萄糖耐量、Ins及高密度脂蛋白（HDL）均显著降低（P<0.01）;与DM组比较,NJ组与NT组小鼠GSP、TG及LDL均明显降低（P<0.01,P<0.05）,葡萄糖耐量、Ins和HDL明显升高（P<0.05）。结论：诺丽果汁与纳豆具有降低糖尿病模型小鼠血糖、增加糖耐量及改善血脂的作用,提示二者对糖尿病防治可能具有一定的应用价值。     

Adrenaline potentiates antiepileptic but not sedative action of diazepam in rats

Intramuscular (i.m.) administration ofdiazepam in a dose of 10 mg/kg and adrenaline in a dose of 0.2 mg/kg prevents generalized clonic-tonic pentylenetetrazol (PTZ) seizures in 75-80 % of rats, but only in 35-40 % of rats it prevents local clonic PTZ seizures. In the above mentioned dose, diazepam causes a strong sedation, but adrenaline does not cause a sedative effects. The combined administration of diazepam and adrenaline in threshold independently ineffective doses prevents both clonic-tonic and clonic PTZ seizures in 80 % of rats without a sedation development. The basis for mechanism of potentiation of anticonvulsant action of diazepam is the stimulation of gastric mucosa afferents by adrenaline.     

Embryotoxicity of oral administered chlorothalonil in mice

BACKGROUND: Chlorothalonil (2,4,5,6-tetrachloroisophthalonitril), the nephrotoxic fungicide, was examined for its potential to produce developmental toxicity in mice after oral administration. METHODS: Pregnant ICR (CD-1) mice were given sublethal doses of 0 (corn oil), 100, 400, and 600 mg/kg/day chlorothalonil by gavage on gestation days (GD) 6-15. RESULTS: Maternal effects in 400 and 600 mg/kg/day dose groups included signs of toxicity such as weakness and depression in the maternal activity, and reduction in body weight and weight gain. No maternal toxicity was apparent in the 100 mg/kg/day dose group. Maternal exposure to chlorothalonil during organogenesis significantly affected the number of live fetuses, early resorption, and mean fetal weight in the 400 and 600 mg/kg/day dose groups. No external, visceral, and skeletal abnormalities were observed among any of the treated groups compared to the control. CONCLUSIONS: On the basis of the present results chlorothalonil can produce clinical signs of toxicity and fetotoxicity without teratogenic effects at 400 and 600 mg/kg/day dose groups.     

新加良附方对移植性小鼠肝癌抑制率及其凋亡影响研究

目的:观察新加良附方对移植性小鼠肝癌(H22)生长抑制作用及其对肿瘤组织中Bcl-2和Bax表达影响。方法:建立移植型H22动物模型,并将动物模型随机分模型对照、环磷酰胺(CTX)与新加良附方大、中、小剂量5组。新加良附大、中、小剂量组给药量分别为10g/kg、5g/kg和2.5g/kg;CTX组给药计量为17mg/kg;模型组给予等量无菌生理盐水。连续给药、给水12天处死模型小鼠分离肿瘤,检测肿瘤大小、称重计算肿瘤抑制率,并将肿瘤组织切片,免疫组化法检测Bcl-2和Bax基因。结果:新加良附大剂量组肿瘤抑制率为48.5%,与模型对照组比较,有统计学意义(P<0.01);新加良附大、中剂量可降低肿瘤组织中Bcl-2蛋白基因表达,升高Bax蛋白基因表达,与模型组比较,有显著性差异(P<0.01,P<0.05)。结论:新加良附方可抑制H22瘤体生长,且下调Bcl-2蛋白基因表达,上调Bax基因,提示新加良附方在抗肿瘤方面具有进一步深入研究价值。     

Protective effect of Withania somnifera Dunal on the behavioral and biochemical alterations in sleep-disturbed mice (Grid over water suspended method)

Sleep disruption involves extensive changes in physiological function, including EEG, motor, metabolic, autonomic processes physiological homeostasis and psychological balance that are necessary for physical health. Benzodiazepines are the most widely used drugs for the sleep related problems in spite of their limitations and side effects. Objective of the study was to investigate the protective effect of W. somnifera on the behavioral and biochemical alterations in sleep disturbed mice. Pretreatment with W. somnifera root extract (100. 200 mg/kg) and diazepam (0.5 mg/kg) significantly protected reduction in body weight, improved the reduced locomotor activity and anxiety levels in animals. Biochemical studies also revealed that W. somnifera (100 and 200 mg/kg) and diazepam (0.5 mg/kg) pretreatment for five days decreased significantly lipid peroxidation, nitrites levels and improved catalase, and reduced glutathione levels. Co-administration of W. somnifera (100 mg/kg) with diazepam (0.5 mg/kg) improved significantly all the biochemical parameters as compared to their effect per se. Preliminary results suggest that Withania root extract can be used in the management sleep loss and associated oxidative stress.