龙芽楤木皂苷对培养心肌细胞钠通道电流的影响

目的:在新生鼠心肌细胞培养液中加入龙芽楤木皂苷(Aralosides As)后,对细胞膜上钠离子流产生的效应进行观察。方法:采用全细胞膜片钳技术,观察不同浓度的As(1μg/ml,10μg/ml,100μg/ml)对培养大鼠心室肌细胞钠电流(INa)幅值的影响。结果:1)As使培养心室肌细胞的INa电流-电压(I-V)曲线明显下移,原有的(I-V)依赖特征不变。2)As在10μg/ml,100μg/ml浓度时,均明显剂量依赖性增加INa的峰值,分别使峰值增加29%和41%。结论:As增加培养心肌细胞INa幅值,可能是实现其正性肌力的作用机制之一。     

竹节香附素A的体外抗肿瘤活性研究

目的:研究两头尖中主要有效成分竹节香附素A的体外抗肿瘤活性,为两头尖抗肿瘤有效成分竹节香附素A的创新药物研究提供科学依据。方法:采用体外MTT法,研究竹节香附素A对人体LAX肺癌、QGY-7703肝癌、Bre-04乳腺癌、Col-6肠癌及L929小鼠成纤维细胞株的增殖抑制作用;考察竹节香附素A对QGY-7703和L929细胞增殖抑制的量-效和时-效关系。采用流式细胞仪研究竹节香附素A对QGY-7703肿瘤细胞的诱导凋亡作用。结果:竹节香附素A对QGY-7703和L929细胞具有显著的抑制增殖作用,且其增殖抑制作用与药物给药浓度、给药时间呈现明显的量效和时效关系。竹节香附素A作用48小时后,对肝癌QGY-7703、肺癌LAX、乳腺癌Bre-04、肠癌Col-6和小鼠成纤维L929五种肿瘤细胞均具有抑制增殖作用,IC50分别为:5.09μg/ml,11.48μg/ml,9.32μg/ml,11.22μg/ml,12.71μg/ml,顺铂的IC50为:5.27μg/ml,6.04μg/ml,6.23μg/ml,6.78μg/ml和9.75μg/ml,其中竹节香附素A对QGY-7703细胞增殖抑制作用最强,作用强度略高于顺铂。竹节香附素A可以显著诱导QGY-7703细胞凋亡,给药剂量的增加,凋亡细胞比例增大。竹节香附素A低剂量(1μg/ml)给药,QGY-7703细胞的凋亡率为19.78%高于顺铂(2μg/ml)给药的细胞凋亡率12.68%,竹节香附素A给药(50μg/ml)24h细胞凋亡率达到67.29%。结论:首次系统研究了竹节香附素A的体外抗肿瘤活性,竹节香附素A体外对QGY-7703具有显著的抑制作用,进而发挥诱导QGY-7703凋亡的生物活性。     

β-细辛醚对PC12细胞和乳鼠皮层神经细胞形态学及细胞活力的影响

目的:观察β-细辛醚对PC12细胞和乳鼠皮层神经细胞形态学及细胞活力的影响。方法:用免疫细胞化学方法检测皮层神经细胞特异性标志物NSE、GFAP的表达;在体外将不同浓度β-细辛醚与PC12细胞和皮层神经细胞各作用24 h,用相差显微镜观察细胞形态的改变,并用MTT比色法检测细胞活力的变化。结果:原代培养的乳鼠皮层神经细胞大多数NSE呈阳性表达,少量GFAP呈阳性表达;β-细辛醚与PC12细胞作用24 h,低浓度β-细辛醚(7.5、15、30、60μg/ml)有促增殖作用,高浓度β-细辛醚(120、140、2480μg/ml)则可抑制其增殖,且随药物浓度的增加而增强;β-细辛醚与皮层神经细胞作用24 h,7.5、15、30、60、120μg/mlβ-细辛醚对其细胞形态和活力无明显影响,240μg/mlβ-细辛醚对其有促增殖作用,而480μg/mlβ-细辛醚对其则有损伤作用。结论:β-细辛醚可能有抗肿瘤和保护神经细胞的作用。     

补肾活血方有效组分及部分化合物的体外抗炎活性研究

目的:观察补肾活血方水提取部位、醇提取部位、总多糖部位以及大黄酸、大黄酚等有效组分与化合物对肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)及一氧化氮(NO)等炎症介质释放浓度的影响作用,探讨复方治疗骨关节炎的初步机制。方法:选取补肾活血方不同提取分离部位,以及复方水提液中存在的具有潜在治疗骨关节炎活性的大黄酸、大黄酚化合物,采用体外脂多糖(LPS)刺激的RAW264.74小鼠巨噬细胞为模型,利用酶联免疫法(ELISA)测定试药对模型细胞释放TNF-α、IL-6及NO的抑制作用。结果:复方水提取部位与醇提取部位在100μg/ml,以及大黄酚在20μg/ml和100μg/ml对TNF-α的释放具有显著的抑制;复方醇提取部位在100μg/ml,大黄酸在20μg/ml,以及大黄酚在20μg/ml和100μg/ml对IL-6水平具有显著的抑制;复方水提取部位在100μg/ml、醇提取部位在20μg/ml和100μg/ml、大黄酸在20μg/ml,以及大黄酚在20μg/ml和100μg/ml对NO的释放具有显著抑制。结论:补肾活血方的治疗骨关节炎活性可能是通过抑制炎症介质释放,降低TNF-α、IL-6、NO等含量发挥作用,这对验证复方药效活性,探究效应物质基础具有重要意义。     

药刀的使用与保养

生牛膝(Ⅰ)、酒牛膝(Ⅱ)、盐牛膝(Ⅲ)醚提物对EB病毒激活的最低浓度分别为0.8,20,0.16μg/ml;5μl醚提物(相当于250mg生药)对鼠耳皮肤有致炎作用;I的醚提物在0.04～125mg/ml对CHL细胞对染色体畸变率无明显影响;小鼠灌胃15g/kgx3d,孕鼠灌胃10g/kgx10d,Ⅰ,Ⅱ,Ⅲ对小鼠骨髓微核率及早孕率无明显影响。     

炮制对芫花中芫花酯甲含量的影响

生牛膝(Ⅰ)、酒牛膝(Ⅱ)、盐牛膝(Ⅲ)醚提物对EB病毒激活的最低浓度分别为0.8,20,0.16μg/ml;5μl醚提物(相当于250mg生药)对鼠耳皮肤有致炎作用;I的醚提物在0.04～125mg/ml对CHL细胞对染色体畸变率无明显影响;小鼠灌胃15g/kgx3d,孕鼠灌胃10g/kgx10d,Ⅰ,Ⅱ,Ⅲ对小鼠骨髓微核率及早孕率无明显影响。     

黄芪多糖、桂皮醛、川芎嗪对实验性糖尿病大鼠创面成纤维细胞增殖作用的影响

目的:探讨黄芪多糖(astragalus polysaccharides,AP)、桂皮醛(japanese cinnamon bark aldehyde,JA)、川芎嗪(ligustrazine,LZ)对实验性糖尿病大鼠创面成纤维细胞增殖的影响.方法:利用MTT法观察不同浓度黄芪多糖、桂皮醛、川芎嗪单体及药物有效浓度之间的不同配伍,在不同时间点上对实验性糖尿病大鼠创面成纤维细胞增殖的影响.结果:发现黄芪多糖浓度为625μg/ml、2.44μg/ml,川芎嗪浓度为50 0μg/ml、125μg/ml时可抑制体外培养成纤维细胞的生长,而桂皮醛浓度在2.8μg/ml、1 .4μg/ml时无明显促进或抑制作用.黄芪多糖浓度为39μg/ml、9.75μg/ml、2.44μg/m l,桂皮醛浓度为22.4μg/ml、11.2μg/ml、5.6μg/ml,川芎嗪浓度为31μg/ml、7.75 μg/ml、1.94μg/ml时可促进成纤维细胞的增殖,并且其增殖作用随时间的延长而增强. 黄芪多糖、桂皮醛、川芎嗪三种单体有效浓度之间配伍,在黄芪多糖和川芎嗪的配伍浓度为 187μg/ml、46.75μg/ml、11.69μg/ml,川芎嗪和桂皮醛的配伍浓度为53.4μg/ml、18 .95μg/ml、7.54μg/ml,黄芪多糖和桂皮醛的配伍浓度为178.4μg/ml、50.2μg/ml、15.35μg/ml,黄芪多糖、桂皮醛和川芎嗪三者配伍浓度为209.4μg/ml、57.95μg/ml、 17.29μg/ml时可促进成纤维细胞的增殖,并且其增殖作用随时间的延长而增强.结论:黄芪多糖、桂皮醛和川芎嗪三种单体联合应用时,对实验性糖尿病大鼠创面成纤维细胞增殖作用优于黄芪多糖、桂皮醛和川芎嗪单独应用(P<0.01)及两种单体联合应用(P<0.05).     

龙牙葱木皂苷对培养心肌细胞游离钙浓度的影响

目的:观察龙牙葱木皂苷对培养心室肌细胞游离钙离子浓度的影响,旨在从细胞水平进一步证明了As的正性肌力作用的机制。方法:采用特异性的荧光指示剂Fluo-3/AM(5μmmol/l)负载心肌细胞,应用激光共聚焦显微镜观察并连续动态扫描细胞内荧光强度的变化,从而反映其对细胞内游离钙浓度的影响。结果:As(10μg/ml)明显增加培养乳鼠心肌细胞内的游离钙离子浓度。Ca2+平均荧光强度明显升高,与对照组相比,P<0.01,差异显著。荧光强度比正常对照组增30%。结论:增加心肌细胞的钙离子浓度,影响心肌兴奋-收缩藕过程,As增加心肌细胞的收缩力,进而实现正性肌力作用。     

炮制对芫花中芫花酯甲含量的影响

生牛膝(Ⅰ)、酒牛膝(Ⅱ)、盐牛膝(Ⅲ)醚提物对EB病毒激活的最低浓度分别为0.8,20,0.16μg/ml;5μl醚提物(相当于250mg生药)对鼠耳皮肤有致炎作用;I的醚提物在0.04～125mg/ml对CHL细胞对染色体畸变率无明显影响;小鼠灌胃15g/kgx3d,孕鼠灌胃10g/kgx10d,Ⅰ,Ⅱ,Ⅲ对小鼠骨髓微核率及早孕率无明显影响。     

人参二醇组皂甙与人参三醇组皂甙对培养心肌细胞钙通道的阻滞作用

PDS与PTS使培养的心肌细胞动作电位各参数呈剂量依赖性减小。PDS 1000肛g/ml、PTS200μg/ml与钙通道阻滞剂Mn~(2+) 0.05μg/ml的作用相当。洗脱、肾上腺素10μg/ml、Ca~(2+)80μg/ml使动作电位的抑制逆转。表明PDS与PTS有钙通道阻滞作用。     

The direct negative inotropic effect of gossypol

Gossypol, an effective male antifertility agent, had a minor but significant negative inotropic action in isolated ferret right ventricular papillary muscles at concentrations of 1-5 micrograms/ml. Reduction of peak developed force and maximum extent of shortening was readily demonstrable at three different resting control contractile states achieved through increases in extracellular calcium from 0.625 to 2.5 mM/l. Gossypol had no significant effect on maximum rate of tension rise nor on maximum velocity of shortening. These results suggest that gossypol reduces the total number of crossbridges without significantly influencing the rate of making and breaking crossbridges and that its negative inotropic action is probably mediated through interactions at the myocardial cell surface.     

Ca2+ channel blocking effect of panaxadiol saponins and panaxatriol saponins of cultured cardiac cells]

It was found that PDS and PTS decreased the action potential parameters of the cultured myocardiocytes in a dosage-dependent way. The effects of PDS 1000 micrograms/ml, PTS 200 micrograms/ml were in correspondence with the known Ca2+ channel blocker Mn2+ 0.05 micrograms/ml. Washing out, administration of epinephrine 10 micrograms/ml or Ca2+ 80 micrograms/ml were able to reverse the action potential from inhibition. The above results indicate that both PDS and PTS have Ca2+ channel blockade action.     

Pharmacology of an Indian-snuff obtained from Amazonian Maquira sclerophylla

The powdered bark of Maquira sclerophylla is consumed as snuff in north Brazil. Both the crude and the purified hydrosoluble extract (WP) injected i.p. in the dose range of 0.05-0.5 g/kg induced hyperexcitability, tremors, motor incoordination, ataxia, quietness and muscle relaxation in rats. The effects were progressive, dose-related and reversed after 30 min. Anesthetized rats, guinea-pigs and dogs injected with the purified extract (10-50 mg/kg, i.v.) showed a biphasic change of carotid blood pressure. The early and transient hypotension was blocked by atropine but not by vagotomy: the secondary hypertension was long lasting and sustained for over 30 min. The hypertension was shortened but not blocked after ganglionic blockade or reserpine treatment. Either pithing or alpha receptor blockade with yohimbine reduced both effects of the extract. Guinea-pigs and dogs were more responsive than rats and died by heart arrest. Incubation of WP (20 micrograms/ml) increased both the rate and force of contraction of isolated guinea-pig right atria by 2 and 5 times, respectively. Propranolol (4 micrograms/ml) blocked the chronotropic effect but did not decrease the inotropic effect. In electrically driven guinea-pig left atria, WP (10 micrograms/ml) increased the force of contraction by 80% and the maximum rate of force development by 60%, but did not change the time to peak tension, the time to 50% relaxation, or the rate of relaxation. These cardiovascular effects resemble those of digitalis-like drugs. Cardenolides were detected in WP by phytochemical screening.     

Studies on cardiovascular actions of Salvia miltiorrhiza

Cardiovascular actions of S. miltiorrhiza (SM) were studied on systemic blood pressure in the rat. Langendorff cardiac preparation in the guinea pig, and four types of vasculature in the dog, including coronary, renal, femoral, and mesenteric arteries. SM induced dose-related hypotension without changing heart rate. The hypotension was antagonized by atropine, propranolol, and chlorpheniramine plus cimetidine. In the isolated whole-heart preparation, SM increased coronary blood flow significantly for 15 min and positive inotropic action for 3 min after pulse injection. SM relaxed all arteries at low concentration (3.0 mg/ml) and contracted all but the coronary artery at higher concentration (10.0 mg/ml). The coronary artery relaxed at all doses of SM tested.     

桂郁金醇提物和水提物对离体蛙心的影响

目的 探讨桂郁金醇提物和水提物对离体蛙心的影响.方法 采用斯氏蛙心灌流法制备离体蛙心标本,逐滴加入桂郁金提取物,观察药物对离体蛙心的作用.结果 当桂郁金醇提物和水提物的终浓度分别达到2.54×10-2g/ml和2.55×10-3g/ml时,开始明显抑制蛙心的收缩张力最大值;两种提取物的终浓度分别达到4.90×10-2g/ml和1.73×10-3g/ml时,开始明显抑制蛙心的收缩张力平均值,并表现出明显的量效关系.两种提取物对蛙心收缩频率的影响并不明显.药物溶媒(纯净水)对蛙心没有明显影响.结论 桂郁金醇提物和水提物有显著的负性肌力作用.     

中药活性成分体外逆转肿瘤细胞多药耐药性的研究

目的：从中药活性成分中筛选肿瘤耐药逆转剂,并对其逆转作用进行研究。方法：采用MTT法测定药物的体外杀伤作用,应用金氏公式进行联合用药分析。结果：钩藤总碱5μg/ml、药根碱2μg/ml、肯定玉红1．25μg/ml对长春新碱在KBv200细胞的逆转倍数分别为16．8、5．1和4倍;姜黄素1．56－12．5μg/ml与长春新碱合用对KB和KBv200细胞均具增敏作用。结论：钩藤总碱、药根碱、靛玉红均可逆转kBv200细胞对长春新碱的耐药性,而姜黄素与长春新碱合用对KB和KBv200细胞均具增敏作用。     

定量PCR检测黄芪A6组分与无环鸟苷联合抗1型单纯疱疹病毒的协同作用

目的：探讨抗疱疹病毒药物黄芪Ａ６组分（Ａ６）和无环鸟苷（ＡＣＶ）联合抗１型单纯疱疹病毒（ＨＳＶ１）的作用机制。方法：利用竞争ＰＣＲ检测Ａ６和ＡＣＶ联合抗ＨＳＶ１的协同作用，并与细胞病变（ＣＰＥ０抑制法进行比较。结果：竞争ＰＣＲ测定Ａ６、ＡＣＶ和Ａ６＿ＡＣＶ对ＨＳＶ１的最小抑制浓度（ＭＩＣ）分别为１．８８ｍｇ／ｍｌ、３．３７μｇ／ｍｌ、３．３７μｇ／ｍｌ和０．４７ｍｇ／ｍｌ＋０．８４μｇ／ｍｌ；ＣＰ     

Effects of gossypol acetic acid on rat luteal cells in vitro

The action of gossypol acetic acid (GAA) on 125I-hCG binding, gonadotropin-stimulated cAMP accumulation and progesterone production was investigated utilizing rat ovaries. Incubation of luteal cells for 3 h with increasing concentration of GAA caused a significant inhibition of gonadotropin-stimulated steroidogenesis. The inhibitory effect of GAA was concentration dependent. GAA at concentrations of 10-30 micrograms/ml reduced cAMP formation in response to hCG. It was shown that the activity of adenylate cyclase of luteal cells was inhibited by 10 micrograms/ml GAA. GAA at a concentration of 30 micrograms/ml was found to have an inhibitory effect on 8Br-cAMP-stimulated progesterone production. GAA did not affect 125I-hCG binding to LH receptor on the luteal cell surface. These results suggest that in luteal cells GAA inhibits steroidogenesis at the step of gonadotropin-stimulated cAMP formation. Adenylate cyclase of luteal cells was inhibited.     

Autonomic nervous system mediates the cardiovascular effects of Rhodiola sacra radix in rats

ETHNOPHARMACOLOGICAL RELEVANCE: Rhodiola sacra (Crassulaceae) exhibits cardiovascular bioactivities and is used in Tibetan medicine for promoting circulation and preventing hypertension. However, the underlying mechanisms of its cardiovascular effects are poorly understood. AIM OF THE STUDY: The aim of this study was therefore to evaluate the cardiovascular activity of water-soluble fraction (WtF) and n-butanol-soluble fraction (BtF) of Rhodiola sacra radix and to explore its mechanism of action in propofol anesthetized Sprague-Dawley rats. MATERIALS AND METHODS: The changes of blood pressure, heart rate and cardiac contractility after systemic administration of the extracts (10-75mg/kg) were examined for at least 40min. Different antagonists were used to evaluate the mechanisms of cardiovascular effects of the extracts. RESULTS: Intravenous injection of the WtF (10, 25, 35, 50 or 75mg/kg) exhibited dose-dependent hypotension and increases in heart rate and cardiac contractility. In contrast, mild alterations in the same cardiovascular parameters were detected only at high dose (75mg/kg) BtF. The WtF-induced hypotensive, positive inotropic and chronotropic effects were significantly abolished by pretreatment with hexamethonium (30mg/kg, i.v.) or reserpine (5mg/kg, i.v.), whereas the hypotensive, but not the positive inotropic or chronotropic effect was potentiated by captopril (2.5mg/kg, i.v.). Pretreatment with methylatropine (1mg/kg, i.v.), on the other hand, reversed the positive inotropic and chronotropic but not the hypotensive effects of WtF. The WtF-induced cardiovascular responses were not affected in rats pretreated with N(G)-nitro-l-arginine methyl ester (20mg/kg, i.v.). CONCLUSIONS: We conclude that systemic administration of the WtF of Rhodiola sacra radix elicited a potent hypotensive effect that was mediated by the withdrawal of sympathetic vasomotor tone and interaction with the circulatory angiotensin system. The positive inotropic and chronotropic effects of WtF may result from a direct vagal inhibition on the heart.     

A drug used in traditional medicine: Harpagophytum procumbens DC II. Cardiovascular activity

In conscious normotensive rats the dried crude methanolic extract of Harpagophytum procumbens secondary roots caused a significant dosedependent reduction of arterial blood pressure. The decrease was significant only at higher doses given by gavage (dried extract = 400 $\text{mg}\text{kg}$). At the same time a decrease of heart rate was observed.In the same experimental conditions, harpagoside presented an activity lower than doses of Harpagophytum procumbens extract containing corresponding quantities of harpagoside.In spontaneously beating Langendorff preparations of rabbit heart, the Harpagophytum procumbens methanolic extract caused a mild decrease in the heart rate with a concomitant mild positive inotropic effect at lower doses but a marked negative inotropic effect at higher doses. The coronary flow decreased at higher doses only.The negative chronotropic and positive inotropic effects of harpagoside were comparatively higher with respect to that of the extract, whereas harpagide had only a slight negative chronotropic effect and a considerable negative inotropic one.Both in experiments on intact rats and on isolated rabbit heart, the Harpagophytum procumbens extract also demonstrated a protective action with regard to arrhythmias induced by aconitine, and particularly to those provoked by calcium chloride and epinephrine—chloroform.