γ-氨基丁酸受体介导的四逆散改善睡眠作用

目的:基于γ-氨基丁酸-苯二氮卓类受体探讨四逆散改善睡眠作用机制。方法:观察了四逆散联合苯二氮卓类受体阻断剂氟马西尼对戊巴比妥钠所致小鼠睡眠时间的影响,以及四逆散对γ-氨基丁酸-苯二氮卓类受体al和a5亚单位mRNA表达的影响。结果:四逆散1.25g/kg连续给药7天,可以明显延长戊巴比妥钠所致小鼠的睡眠持续时间和缩短其潜伏期。氟马西尼3.5mg/kg可以明显阻断四逆散1.25g/kg对戊巴比妥钠所致小鼠睡眠的增强作用。四逆散能使小鼠大脑皮质γ-氨基丁酸A受体a5亚单位的mRNA表达明显上调,有效剂量为1.25g/kg,给药起效时间为7天,给药9天效果较显著。结论:γ-氨基丁酸-苯二氮卓类受体介导四逆散改善睡眠作用。     

四物汤抗利多卡因毒性的研究

四物汤５ｇ／ｋｇｉｐ能显著对抗利多卡因的毒性，并增加大鼠对利多卡因的耐受量；１．７ｇ／ｋｇｉｖ能降低利多卡因中毒小鼠的死亡率。     

半夏对大白鼠妊娠及胚胎的毒性研究

半夏对大白鼠妊娠和胚胎的毒性试验表明:生半夏粉9 g/kg 灌胃,对妊娠母鼠和胚胎均有非常显著的毒性,而相同剂量的制半夏粉与对照组无明显的差异,但是制半夏汤剂30g/kg(相当于临床常用量的150倍)则能引起孕鼠阴道出血,胚胎早期死亡数增加,胎儿体重显著降低,提示前人所谓"半夏动胎"是有其道理的生半夏汤剂30g/kg 对大白鼠妊娠和胚胎的毒性与制半夏汤剂无差异,说明半夏汤剂的毒性不因炮制而降低。     

基于NO信号通路介导的四逆散改善睡眠作用的机制研究

目的：探究四逆散改善睡眠的作用机制。方法：采用化学比色法,观察四逆散冻干粉2500 mg/kg对小鼠脑内NO含量和一氧化氮合酶（TNOS）活力的影响。应用戊巴比妥钠所致小鼠睡眠的实验方法,观察小鼠给予不同剂量四逆散冻干粉合并L-精氨酸（L-Arg）,以及四逆散冻干粉合并N-硝基-L-精氨酸甲酯（L-NAME）后小鼠入睡潜伏期和睡眠持续时间。结果：与空白组比较,四逆散冻干粉2500 mg/kg能明显增加小鼠全脑的NO含量（P〈0.05）和TNOS的活性（P〈0.01）。L-Arg 500 mg/kg表现为延长戊巴比妥钠所致小鼠的睡眠时间（P〈0.05）,剂量为1000 mg/kg时,主要表现为明显缩短戊巴比妥钠所致小鼠的睡眠时间（P〈0.05）和延长其潜伏期（P〈0.01）;L-Arg 125 mg/kg可延长给予四逆散冻干粉625 mg/kg的小鼠睡眠持续时间（P〈0.01）和缩短睡眠潜伏期（P〈0.05）;L-Arg 500mg/kg可缩短给予四逆散冻干粉2500 mg/kg的小鼠睡眠持续时间（P〈0.05）和延长睡眠潜伏期（P〈0.01）;L-NAME（40 mg/kg或80 mg/kg,腹腔注射）能明显延长戊巴比妥钠所致小鼠的睡眠持续时间（P〈0.05）和缩短其潜伏期（P〈0.05）。L-NAME 80mg/kg可显著缩短给予四逆散冻干粉的小鼠睡眠持续时间（P〈0.01）。结论：NO介导参与四逆散改善睡眠作用,四逆散通过增强NOS活力,适量增加NO含量发挥改善睡眠的作用。     

四逆散对HepG2移植性肝癌裸鼠Sirt1和p53表达的影响

目的:探讨四逆散对裸鼠移植性肝癌的抑制作用。方法:利用HepG2细胞裸鼠移植瘤模型,观察四逆散的抗肿瘤作用,采用ELISA法测定白介素-6(IL-6)、白介素-12(IL-12)和肿瘤坏死因子-α(TNF-α)的水平,并用Western blot法探讨其作用机制。结果:与模型组比较,四逆散(1.8⁷.2g/kg)对荷瘤小鼠肿瘤的生长具有明显的抑制作用,四逆散中(3.6g/kg)剂量组和高(7.2g/kg)剂量组能够明显增加活性氧(ROS)的含量,并提高小鼠脾脏指数、IL-6、IL-12和TNF-α的水平,同时,显著下调Bcl-2、Sirt1蛋白表达,上调Bax、Caspase 3和p53的蛋白表达。结论:四逆散能够抑制人肝癌细胞HepG2裸鼠移植瘤生长,并通过ROS依赖的Sirt1-p53凋亡途径发挥作用。     

结肠炎丸的药效学研究

结肠炎丸8g/kg、4g/kg及2g/kg能明显增加大黄型脾虚小鼠的体重,显著减少其稀便次数;8g/kg及4g/kg结肠炎丸显著提高RES吞噬功能,同时明显提高血清溶血素的生成。     

复方丹参片的药理作用研究

实验结果表明,给动物口服复方丹参片10g/kg,有明显抑制氯仿引起小鼠心室纤颤作用。静脉注射0.4g/kg,有缩短氯化钡引起的大鼠心律失常时间的作用。口服3.5g/kg和腹腔注射13.5g/kg,均能明显延长小鼠常压耐缺氧时间,并能显著对抗垂体后叶素引起的心电图T波变化。复方丹参片液0.1ml(相当于成人每次口服量的3倍)和0.2ml(相当于成人每次口服量的6倍)均有显著增加冠脉流量的作用,并随着剂量的增加而作用增强。复方丹参片对缺氧灌流液所致心衰的冠脉流量增加18.6%,口服10g/kg能显著延长小鼠的凝血时间。     

高乌头炮制前后毒性部位与镇痛抗炎药效部位筛选研究

目的:筛选高乌头炮制前后毒性及药效部位,探讨毒性与药效的关系,为高乌头减毒存效合理炮制提供科学依据。方法:采用小鼠急性毒性实验比较不同溶剂萃取部分的毒性,筛选生、制高乌头的毒性部位,以热板法、扭体法及二甲苯致鼠耳肿胀实验考察不同剂量生、制高乌头毒性部分对小鼠的镇痛抗炎作用;筛选药效部位,探究毒性部位是否为药效部位。结果:生、制高乌头均为三氯甲烷部分的毒性最强,水提及正丁醇部分次之,生品LD50分别为0.09、1.81、24.41 g/kg,制品LD50分别为0.11、2.42、0.87 g/kg;而石油醚、乙酸乙酯及乙醇部分毒性均最小,生品最大给药量分别为2.69、3.11、28.38 g/kg,制品最大给药量分别为13.12、1.98、20.32 g/kg。给药后120 min、1.5、4.5 mg/kg的生品三氯甲烷部分与4.5 mg/kg的制品三氯甲烷部分均能提高小鼠热板痛阈值,且呈量效关系,镇痛作用优于1.6 mg/kg氢溴酸高乌甲素组。注射冰醋酸15~30 min,1.5、4.5 mg/kg的生、制品三氯甲烷部分均能有效抑制冰醋酸所致小鼠的扭体次数,且呈量效关系,抑制率略低于1.6 mg/kg氢溴酸高乌甲素组。在二甲苯致鼠耳肿胀实验中,1.5、4.5 mg/kg的生、制高乌头三氯甲烷部分均能有效缓解小鼠耳肿胀度,且呈量效关系,肿胀抑制率略低于10.0 mg/kg消炎痛组。结论:三氯甲烷部分既是生、制高乌头的毒性部位,又是其镇痛抗炎的主要药效部位,且呈量效关系,推测其镇痛抗炎活性与毒性存在一定的关联。     

四逆散有效组分改善睡眠作用的量效、时效关系及作用机制研究

目的:在研究四逆散有效组分改善睡眠作用的量效、时效关系的基础上从NO信号通路探讨四逆散有效组分改善睡眠的作用机制。方法:采用戊巴比妥钠所致小鼠睡眠实验方法,以小鼠睡眠时间为指标研究四逆散有效组分改善睡眠作用的量效、时效关系,四逆散有效组分联合L-精氨酸、N-硝基-L-精氨酸甲酯应用改善睡眠作用,并测定给予四逆散有效组分后小鼠脑干与丘脑的NO含量。结果:四逆散有效组分改善睡眠作用时效关系明显,最短起效时间为连续给药4天,量效关系复杂在给药剂量增加到660.00mg/kg时达到药效高峰,在给药剂量1320.00mg/kg时药效减弱;给予四逆散有效组分后小鼠脑干与丘脑的NO含量增加;联合L-精氨酸给药后,睡眠持续时间相对于四逆散有效组分有延长作用;联合N-硝基-L-精氨酸甲酯给药后,睡眠持续时间相对于四逆散有效组分有缩短作用。结论:四逆散有效组分改善睡眠作用时效关系明显、量效关系复杂,其改善睡眠的作用NO信号通路参与了介导。     

苍耳子炮制前后急性毒性及毒效成分含量差异研究

目的：研究苍耳子炮制前后对小鼠的急性毒性及主要毒性成分、药效成分的含量差异。方法：急性毒性实验采用小鼠灌胃给药测定其半数致死量(LD₅₀)和最大耐受量(MTD),并通过组织病理切片观察其对小鼠肝、肾等脏器的损伤;采用高效液相色谱法比较苍耳子炮制前后2种水溶性苷类毒性成分和4种酚酸类有效成分的含量差异。结果：苍耳子的水煎剂和水煎剂醇沉沉淀,以及炒苍耳子的水煎剂和水煎剂醇沉沉淀的MTD分别为291.73 g生药/kg、405.75 g生药/kg、318.64 g生药/kg和480.77 g生药/kg。炮制前后苍耳子水煎剂醇沉上清的LD₅₀分别为468.18 g生药/kg和573.48 g生药/kg。组织病理结果显示死亡小鼠多数脏器出现不同程度的充血,以肝、肺、胃和小肠最为明显。苍耳子炮制后,水煎剂和水煎剂醇沉上清液中的羧基苍术苷含量均显著降低(均P<0.01);炒苍耳子中咖啡酸和异绿原酸C的含量显著升高(均P<0.01)。结论：苍耳子炮制后毒性可显著降低,有效成分溶出增加。水煎剂醇沉上清液取自苍耳子的主要毒性部位。     

Pharmacological study on the compatibility of cortex Cinnamomi with Halloysitum Rubrum]

The decoction of Cortex Cinnamomi (CC, 1 g/kg p.o.) and Halloysitum Rubrum (HR, 3 g/kg p.o.) or the combination of the two drugs (4 g/kg p.o., CC 1 g/kg, HR 3 g/kg) could antagonize the diarrhea caused by p.o. water ex tract of Radix et Rhizoma Rhei in mice; and inhibit the platelet aggregation induced by ADP in vitro. Meanwhile, the effect of the combination of the two drugs was not different from that of each single one. In addition, CC was able to inhibit the spontaneous movement of intestine in situ and showed an analgesic effect (hot-plate method) in mice; HR was ineffective in these aspects and did not reduce the effect of CC. CC(20 g/kg p.o., i.p. or i.v.) exhibited very strong toxicity in mice, while HR(60 g/kg p.o., i.p. or i.v.) was nontoxic. When the two drugs were used together, the toxicity was markedly reduced.     

调肝、补肾、健脾方药对慢性心理应激大鼠单胺类神经递质影响的比较研究

目的：观察慢性心理应激反应大鼠血浆如下丘脑多巴胺（DA）、5－羟色胺（5－HT）、肾上腺素（E）、去甲肾上腺素（NE）的变化及调肝、补肾、健脾方药对其影响。方法：Wistar大鼠按体重随机分为6组，即对照组（C），模型组（M），调肝中药（加味四逆散）组（C1），人参总皂甙组9C2），补肾中药（肾气丸）组（C3），健脾中药（四君子汤）组（C4）。采用束缚限制活动空间的方法造成慢性心理应激大鼠模型，所有药物灌胃给药。采用电化学高效液相色谱分析方法检测血浆和下丘脑DA、5－HT、E、NE含量。结果：应激大鼠血浆和下丘脑DA、5－HT含量明显降低（均P＜0.01)，血浆NE、E含量明显升高（均P＜0.01)，而下丘脑NE含量无明显变化。血浆5－HT／NE、5－HT／E及下丘脑5－HT／NE均明显降低（均P＜0.01)。加味四逆散可以升高应激大鼠下丘脑和血浆中DA、5－HT含量（均P＜0.01)，并能降低血浆中NE、E含量（均P＜0.01)，升高下丘脑和血浆5－HT/NE以及血浆5－HT／E（P＜0.05或P＜0.01)。肾气丸、四君子汤及人参部 甙虽然对有些指标有影响，但对心理应激的整体调节作用不如加味四逆散。结论：加味四逆散对慢性心理应激大鼠血浆和下丘脑单胺类神经递质的代谢紊乱均有良好的调节作用，调肝、补肾、健脾治法方药中以调肝治法方药的综合调治作用最为显著。     

Oleuropein,Chief Constituent of Olive Leaf Extract,Prevents the Development of Morphine Antinociceptive Tolerance through Inhibition of Morphine‐induced L‐type Calcium Channel Overexpression

It has been shown that blockade of L‐type calcium channels could abolish the development of opioid‐induced antinociceptive tolerance. Here, the antitolerant effects of olive leaf extract (OLE) and its main component, oleuropein, which have a calcium channel blocker property were determined. Adult male Wistar rats were injected with morphine (20 mg/kg, i.p.) for 8 days to induce antinociceptive tolerance. Then OLE (50–200 mg/kg i.g.) and oleuropein (1–10 mg/kg i.p.) were injected concomitantly with morphine. The tail‐flick test was used to assess the nociceptive threshold. The dorsal half of the lumbar spinal cord was assayed for the expression of L‐type calcium channel using semiquantitative RT‐PCR. The results showed that OLE (200 mg/kg) completely prevented morphine tolerance development. In addition, oleuropein in dose of 10 mg/kg, but not in 5 mg/kg, prevented the development of morphine antinociceptive tolerance. In addition, a significant increase in the mRNA levels of calcium channel (43.9%) was observed in the lumbar spinal cord of tolerant animals, which was reversed by effective of dose OLE. In conclusion, the results indicate that olive leaf extract has a potential antitolerant property against the chronic usage of morphine and that its main component, oleuropein, is responsible for such effect. Copyright © 2012 John Wiley & Sons, Ltd.     

Aconitine-induced bradycardia,centrally acting muscarinic effects are inhibited peripherally by higenamine in conscious mice

The effects of aconitine, higenamine and coryneine derived from processed aconite, a Sino-Japanese medicine on pulse rate were examined in unanaesthetized mice. The intraperitoneal (i.p.) administration of the water extract (3 mg/kg) of processed aconite produced a bradycardia, like bigeminy, within 15 min which was prolonged for 30 min. Aconitine (30 μg/kg, i.p.) produced a similar bradycardia. Higenamine induced tachycardia at doses of 30 and 100 μg/kg, i.p. The aconitine-induced bradycardia was antagonized completely by higenamine (10 μg/kg, i.p.), isoproterenol (1 μg/kg. s.c.), or scopolamine (10 μg/kg. s.c.). The anti-aconitine effect of higenamine was prevented by propranolol (1 mg/kg, i.p.). Centrally administered aconitine (1 and 3μg) also induced bradycardia. The aconitine (i.c.v.)-induced bradycardia was prevented by centrally administered atropine (1 μg, i.c.v.). These results demonstrated that aconitine (i.p.)-induced bradycardia is mainly due to a central muscarinic action. As higenamine has a peripheral β₁-adrenergic action, like isoproterenol, the inhibition by higenamine of aconitine-induced bradycardia is due to physiological antagonism.     

Antidiarrhoeal activity of seed extract of Albizzia lebbeck Benth

The antidiarrhoeal activity of the seed extract of Albizzia lebbeck (Benth.) was investigated employing conventional rodent models of diarrhoea, i.e. castor oil-induced diarrhoea, upper gastrointestinal transit (u.g.t.) and fluid secretion. It was found that the aqueous methanol extract of Albizzia lebbeck seeds (2.5-5 mg/kg i.p.) possessed antidiarrhoeal activity which strengthens the earlier use of the seeds in the treatment of diarrhoea and dysentery. The antidiarrhoeal dose of the extract was at least 10-30 times less than the LD(50) dose. The extract (2.5-5 mg/kg i.p.) potentiated the antidiarrhoeal activity of loperamide (1 mg/kg i.p.). Nalaxone (0.5 mg/kg i.p.) significantly inhibited the antidiarrhoeal activity of the extract as well as loperamide, thus indicating a role of the opioid system in the antidiarrhoeal activity of the extract.     

RP-HPLC同时测定四逆散中4种有效成分含量

目的建立同时测定四逆散中芍药苷、柚皮苷、橙皮苷、甘草酸的高效液相色谱方法,为科学评价及有效控制其质量提供可靠方法,并为四逆散的药效物质研究奠定基础。方法采用Kromasil C18柱(4.6 mm×250 mm,5μm),流动相为乙腈-0.05%磷酸水溶液,梯度洗脱,检测波长为240 nm,流速1.0 mL/min,柱温30℃。结果芍药苷、柚皮苷、橙皮苷、甘草酸分别在0.00225~0.01125μg(r=0.999 7)、0.00390~0.00195μg(r=0.9997)、0.00198~0.00099μg(r=0.999 5)、0.00262~0.00131μg(r=0.999 7)范围内线性关系良好,平均加样回收率分别为97.72%(RSD=1.02%)、98.45%(RSD=1.52%)、97.74%(RSD=1.63%)、98.34%(RSD=1.78%)。结论所建立的方法简单、准确、可靠,重复性好,可用于四逆散的质量控制。     

Inhibition of chemically induced inflammation and pain by orally and topically administered leaf extract of Manihot esculenta Crantz in rodents

The aqueous leaf extract of Manihot esculenta Crantz (MELE) is being used orally and topically in traditional African medicine for the treatment of inflammation and pain, and claimed to be safe. The anti-inflammatory effects of MELE (100-400mg/kg, p.o. or 1-4%, w/w in petroleum jelly, topically) were tested against carrageenan-induced paw oedema in rats as well as against xylene-induced ear oedema in mice. The analgesic effect of MELE (100-400mg/kg, p.o. or 1-4%, w/w in petroleum jelly, topically) was tested against acetic acid-induced (20mul, 0.6%, v/v in normal saline, i.p.) and acetylcholine-induced (8.3mg/kg, i.p.) mouse writhing models. At 100-400mg/kg, p.o. and 1-4% (w/w), topically, MELE produced significant inhibitions of carrageenan-induced rat paw oedema and xylene-induced ear swelling in mice. Effects produced by MELE were significantly higher than those produced by indomethacin (10mg/kg, s.c. or 1%, w/w in petroleum jelly) in the anti-inflammatory models. For the analgesic effect, MELE (100-400mg/kg, orally) and (1-4%, w/w, topically), like aspirin (100mg/kg, i.p.) exhibited significant (P<0.05) inhibition of acetic acid- and acetylcholine-induced mouse writhing tests, compared to untreated control. Effects produced by MELE were significantly lower than those produced by aspirin (100mg/kg, i.p.) in the analgesic models, except for the topically administered extract on acetylcholine-induced pain. Acute oral administration up to 10g/kg did not cause death within 14 days, but mortalities were produced in i.p. administered extract with LD(50) of 2.5+/-0.3g/kg. Based on these, the extract may contain orally safe, topically and orally effective anti-inflammatory and analgesic principles, which justify its use in traditional African medicine.     

经方四逆散对肝损害的保护作用及机制研究

[目的]观察经方四逆散对肝损害的保护作用及机制研究。[方法]采用石胆酸灌胃造成肝损害，检测血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、谷胱甘肽(GSH)、超氧化物歧化酶(SOD)、丙二醛(MDA)水平，并作肝组织病理学观察。[结果]经方四逆散能降低石胆酸灌胃造成肝损害小鼠血清ALT、AST、MDA升高的水平，能升高石胆酸灌胃造成肝损害小鼠血清SOD、GSH降低的水平；经方四逆散能使肝损害小鼠肝组织病理变化程度明显减轻。四逆散药粉 米汤降AST、MDA明显，四逆散煎剂降ALT明显。[结论]经方四逆散具有一定的保肝作用。     

四逆散合四君子汤联合西药治疗慢性乙肝肝纤维化随机平行对照研究

[目的]观察四逆散合四君子汤联合西药治疗慢性乙肝肝纤维化疗效。[方法]使用随机平行对照方法,对60例门诊及住院患者按就诊顺序编号随机分为三组(20例/组),均给予西药常规基础治疗,分别加用四逆散、四君子汤及四逆散合四君子汤,连续治疗3个月为1疗程。观测血清肝纤维化指标。连续治疗2疗程(6个月),判定疗效。[结果]血清肝纤维化指标各组均明显改善(P<0.01);四逆散+四君子汤组优于单一四逆散组和四君子汤组(P<0.01);四逆散组和四君子汤组间无显著差异(P>0.05)。[结论]四逆散合四君子汤联合西药可显著改善肝纤维化指标,延缓肝纤维化的进程。