丹皮酚磺酸钠对钙反常培养乳鼠心肌细胞Ca2+内流的抑制作用

培养乳鼠心肌细胞复制钙反常模型,观察丹皮酚磺酸钠对心肌细胞⁴⁵Ca摄取、膜唾液酸(SA)含量及细胞搏动频率的影响,结果提示丹皮酚磺酸钠在50～400μg/ml范围内能显著抑制正常心肌细胞快相(5 min)和慢相(120 min)⁴⁵Ca摄取及其搏动频率,且能显著抑制钙反常心肌细胞的⁴⁵²⁺内流外,尚与其提高膜SA含量有关。     

牛磺酸对大鼠心肌Ca~(2+)调节作用的研究

研究了牛磺酸对大鼠离体左心室肌~(45)Ca内流的影响。对照组~(45)Ca内流与KH液中Ca_-~(2+)浓度有关。当Ca~(2+)浓度分别为0.62(低Ca~(2+)),1.25(正常Ca_-~(2+))和1.87(高Ca_-~(2+))mmol/L时,~(45)Ca内流相应为1.02±0.25,1.37±0.14,和1.45±0.14μmol/g。加入牛磺酸后,上述情况显著改变。低Ca_-~(2+),Tau 10、20、40mmol/L能分别使~(45)Ca内流增加为1.11±0.11,1.45±0.12和1.48±0.09μmol/g:正常Ca~(2+)时,Tau 10、20、40mmol/L能分别使~(45)Ca内流减少为1.19±0.07,1.14±0.23和0.97±0.24μmol/g:高Ca~(2+)时、Tau 10、20、40mmol/L能使~(45)Ca内流显著降低为1.12±0.05,0.58±0.18和0.53±0.10μmol/g。结果表明不同Ca~(2+)浓度能影响心肌~(45)Ca内流,牛磺酸对心肌~(45)Ca内流有双向调节作用,这可能在其抗心律失常机制中起重要作用。     

粉防己碱、蝙蝠葛碱对高K~+引起突触体~(45)Ca内流的影响

<正> 钙离子在中枢神经系统的功能调控及许多病理过程中都有重要作用。而大多数经典钙拮抗剂对神经元Ca~(2+)通道不敏感。最近我们研究表明某些四氢异喹啉类化合物,如蝙蝠葛苏林碱对神经元Ca~(2+)通道有相对选择性拮抗作用。为了进一步确定四氢异喹啉类化合物对神经元Ca~(2+)通道的拮抗活性,我们观察粉防已碱、蝙蝠葛碱对突触体~(45)Ca内流的影响.实验方法Sprague-Dawley大鼠,体重200±23g,雌雄兼用。大鼠断头,迅速分离大脑皮层,制备突触体。用酚试剂法测定蛋白含量,突触体反应浓度为0.9g/L。按文献测定~(45)Ca内流。高K~+(65mmol/L)去极引起”快相”~(45)Ca内流反应时间为20s。净~(45)Ca内流量△(Ca~(2+))i=高K~+引起“快相”~(45)Ca内流—基础~(45)Ca内流。     

甲状旁腺素对心肌细胞Ca~(2+)跨膜转运的影响

应用放射性同位素~(45)Ca示踪技术观察了牛甲状旁腺素_(1-34)活性片段(bPTH_(1-34))对培养乳鼠心肌细胞钙离子(Ca~(2+))跨膜转运的影响.并且镜检了细胞的搏动频率和采用蛋白质竞争结合法检测了胞内cAMP的含量。结果表明:bPTH_(1-34)(1×10~(-7)mol·L~(-1))显著增加静息细胞的搏动频率.明显增高细胞cAMP的含量.并且显著增加静息、高K~+去极化及NA受体兴奋状态细胞的外Ca~(2+)内流.同时抑制胞内Ca~(2+)外排。提示其强心与细胞内Ca~(2+)升高的机理之一.可能与细胞内cAMP升高有关。     

八肽胆囊收缩素对抗吗啡抑制大鼠脑突触小体钙摄取的作用

用大鼠脑的膜制备观察吗啡和CCK-8对突触小体摄取~(45)Ca~(2+)的影响。吗啡(10 nmol/L～1μmo1/L)抑制突触小体对~(45)Ca的摄取,该作用能被1μmol/L纳洛酮完全阻断。CCK-8(10nmol/L～1μmol/L)本身能抑制突触小体~(45)Ca摄取,但它在10nmol/L和100 nmol/L时能对抗吗啡对~(45)Ca摄取的抑制作用,浓度提高到1μmol则不能对抗吗啡的这一作用。CCK-8抑制突触小体摄取~(45)Ca,以及对抗吗啡的~(45)Ca摄取抑制的作用,皆能被CCK受体拮抗剂丙谷酰胺(2μmol/L)所阻断.捉示CCK-8是通过激动CCK受体而拮抗吗啡抑制~(45)Ca摄取的,CCK-8的这一拮抗作用可能是其抗阿片作用的机理之一.     

三七皂甙对血管平滑肌α受体引起~(45)Ca外溢与内流的影响

在血管平滑肌,三七总皂甙明显减少苯肾上腺素引起的~(45)Ca内流量,不影响高钾引起的~(45)Ca内流;而硝苯吡啶几乎完全阻断后一种的~(45)Ca内流。三七总皂甙不抑制~(45)Ca外溢。结果表明,三七总皂甙能特异阻断血管平滑肌α受体操纵的Ca~(2+)通道而不影响胞内Ca~(2+)释放过程。     

何谓钙拮抗剂?目前临床上常用的有哪几种?

<正> 钙拮抗剂的基本作用是阻滞心肌及平滑肌(如冠脉或外周血管平滑肌)细胞的慢Ca~(2+)通道,抑制Ca~(2+)向细胞的内流,故又称钙通道阻滞剂(或抑制剂)。电生理研究证实,心肌细胞动作电位曲线中0相的快速上升是由于Na~+内流所致,钙拮抗剂不影响或很少影响其过程;2相的平台由Ca~(2+)内流所维持,故能被钙拮抗剂所抑制。而属于慢自律细胞的窦房结和房室     

赛庚啶对离体犬心肌肌质网Ca~(2+),Mg~(2+)—ATP酶活性和~(45)Ca~(2+)摄取功能的影响

当赛庚啶浓度在8×10~(-6)mol/L～2×10~(-4)mol/L之间时,该药对正常犬心肌肌质网Ca~(2+),Mg~(2+)—ATP酶活性几乎没有影响,仅在10~(-3)mol/L时对该酶活性才有一定的抑制作用(抑制率为39.85%,P<0.01)。正常犬心肌肌质网的~(45)Ca~(2+)摄取过程有明显的时间依赖性,至第30 min,其~(45)Ca~(2+)摄取量可达312.79±22.25 nmol/mg protein.赛庚啶对心肌肌质网的~(45)Ca~(2+)摄取有一定的抑制作用,其IC_(50)为1.94×10~(-4)mol/L。     

用“冷镧法”测量大鼠用动脉~(45)Ca跨膜流动及对钙拮抗剂的反应

血管平滑肌细胞钙动力学异常引起的血管收缩,是高血压发生的先决条件。近年来为了研究平滑肌细胞外Ca~(++)的跨膜内流,使用~(45)Ca示踪技术是比较好的方法。其中“镧法”更能准确地反映细胞内Ca~(++)水平。我们根据Lewanczuk等的方法略加改动,建立了“冷镧法”测量~(45)Ca细胞摄取     

氯化镉和肝脏镉金属硫蛋白对大鼠肾刷状缘细胞膜的作用比较

本文根据镉中毒肾脏毒性作用的主要表现,通过体外试验研究了CdCl_2及Cd-MT对肾小管刷状缘细胞膜(BBM)的~(45)Ca~(2+)摄取功能及膜表面性质的影响。结果表明:终浓度为1.0—4.0μmol/L的CdCl_2对BBM的~(45)Ca~(2+)摄取功能有显著的抑制作用(P<0.01),而含等量镉的Cd-MT-Ⅰ(Cd:Cu:Zn=1.0:0.8:1.2)和Cd-MT-Ⅱ(Cd:     

高效液相色谱法测定不同产地不同年份牡丹皮中丹皮酚的含量

目的建立高效液相色谱法,测定不同产地、不同年份牡丹皮中丹皮酚含量。方法色谱柱为Diamonsil C18柱（4.6mm×200.0mm,5.0μm）,流动相为甲醇-醋酸-水（54：2：44）,流速1.0ml/min;检测波长274nm。结果丹皮酚在5.1～102.0μg/ml范围内线性关系良好,r=0.9996,平均回收率为99.4%,精密度（RSD）=0.94%。不同产地及同一产地不同年份牡丹皮中丹皮酚含量差异显著（P〈0.01）。结论该方法准确、稳定、简单,可以作为牡丹皮药材的质量控制方法。     

Effect of paeonol on L-type calcium channel in rat ventricular myocytes

In order to study the possible mechanisms of paeonol on the cardiovascular system, the effect of paeonol on L-type Ca(2+) channel current (I(Ca,L)) was studied in rat ventricular myocytes using the whole cell patch-clamp technique. Exposure to paeonol (approximately 10 to 1000 microm/l) resulted in a concentration-dependent inhibition of peak I(Ca,L), with a half maximum inhibition concentration, IC(50), of 561 microm/l. Paeonol 600 microm/l inhibited I(Ca,L) by 55.3%, shifted the steady state activation and inactivation curve of I(Ca,L) to more positive and negative potentials, respectively, tended to prolong the recovery of I(Ca,L) from inactivation and did not have a use-dependent effect. However, the current-voltage relationship and reversal potential of I(Ca,L) were not altered. These results suggest that the protection by paeonol against myocardial injury is due to its blocking effect on I(Ca,L).     

Effect of adrenochrome on calcium accumulation by heart mitochondria

The effects of adrenochrome (1–100 μg/ml or 5.6 × 10^?6 to 5.6 × 10^?4 M) on calcium binding, calcium uptake, and ATPase activities of rat heart mitochondria were investigated. Depression, by adrenochrome, of mitochondrial calcium binding and uptake was observed under in vitro conditions, whereas mitochondrial ATPase activity was not altered appreciably. The inhibitory effect of adrenochrome on calcium uptake activity was independent of the pH of the incubation medium, but it was dependent upon the dose of drug and the time of incubation. High concentrations of calcium in the incubation medium antagonized the adrenochrome-induced depression. The inhibition of mitochondrial calcium uptake by adrenochrome was of a mixed type. Furthermore, the depressed calcium uptake activity was observed after washing adrenochrome-treated mitochondria with buffer. Significant decreases in calcium accumulating activities were also seen in mitochondria isolated from hearts perfused with various concentrations of adrenochrome (5–50 μg/ml) for 10 min, or with 50 μg/ml adrenochrome for various time periods. Contractile force of the perfused rat heart with various concentrations of adrenochrome (5–50, μg/ml) decreased in a dose-dependent manner. It is suggested that the cardiac contractile failure and myocardial cell necrosis induced by adrenochrome may partly be due to its inhibitory effect on the calcium accumulating ability of mitochondria.     

β受体阻滞剂对乳鼠心肌细胞内质网应激致凋亡途径的影响

目的通过体外培养乳鼠心肌细胞,探讨β受体阻滞剂对乳鼠心肌细胞凋亡率及内质网应激作用的影响。方法培养乳鼠心肌细胞,确定β受体阻滞剂的饱和浓度,原代培养乳鼠心肌细胞72h后给予β受体阻滞剂溶液和衣霉素溶液干预,实验分4组:空白对照组、50μg/ml美托洛尔组、10μg/ml衣霉素组、10μg/ml衣霉素+50μg/ml美托洛尔组。确定美托洛尔的饱和浓度;反转录-聚合酶链反应(RT-PCR)检测各组内质网应激指标GRP78、内质网应激致凋亡指标Caspase12,流式细胞术检测实验各组乳鼠心肌细胞凋亡率,确定美托洛尔对内质网应激致凋亡途径的影响作用。结果①美托洛尔的浓度为50μg/ml时,对内质网应激的影响已达最大。②与空白对照组比较,50μg/ml美托洛尔组心肌细胞内质网应激致凋亡指标Caspase12表达及心肌细胞凋亡率差异无统计学意义(P>0.05)。10μg/ml衣霉素组、50μg/ml美托洛尔+10μg/ml衣霉素组心肌细胞内质网应激致凋亡指标Caspase12及心肌细胞凋亡率较空白对照组均明显增加(均P<0.05),且在衣霉素组时心肌细胞中内质网应激致凋亡指标Caspase12及心肌细胞凋亡率均最高,而50μg/ml美托洛尔+10μg/ml衣霉素组与10μg/ml衣霉素组比较,心肌细胞内质网应激致凋亡指标Casepase12及心肌细胞凋亡率降低,差异有统计学意义(均P<0.05)。结论美托洛尔浓度在50μg/ml时为内质网应激保护的饱和浓度,β受体阻滞剂可以减轻内质网应激所致的凋亡途径。     

The effects of paeonol on the electrophysiological properties of cardiac ventricular myocytes

Previous studies have shown that "Mudanpi", a Chinese herbal medicine, has a significant cardioprotective effect against myocardial ischaemia. Based on these findings we hypothesised that paeonol, the main component of Mudanpi, might have an effect on the cellular electrophysiology of cardiac ventricular myocytes. The effects of paeonol on the action potential and ion channels of cardiac ventricular myocytes were studied using the standard whole-cell configuration of the patch-clamp technique. Ventricular myocytes were isolated from the hearts of adult guinea-pig by enzymic dispersion. The myocytes were continuously perfused with various experimental solutions at room temperature and paeonol applied in the perfusate. Action potentials and membrane currents were recorded using both current and voltage clamp modes of the patch-clamp technique. Paeonol, at concentrations 160 microM and 640 microM, decreased the action potential upstroke phase, an action associated with the blockade of the voltage-gated, fast sodium channel. The effects of paeonol on both action potential and Na(+) current were concentration dependent. Paeonol had a high affinity for inactivated sodium channels. Paeonol also shortened the action potential duration, in a manner not associated with the blockade of the calcium current, or the enhancement of potassium currents. These findings suggest that paeonol, and therefore Mudanpi, may possess antiarrhythmic activity, which may confer its cardioprotective effects.     

丹皮酚对IL-1β诱导HFLS-RA的保护作用及相关机制研究

目的　探讨丹皮酚对白细胞介素（IL）-1β诱导人类风湿关节炎成纤维样滑膜细胞（HFLS-RA）的保护作用及机制。方法　将HFLS-RA细胞分为空白对照组,10 μg/L IL-1β组及0.1、1、10、100 μmol/L丹皮酚组,采用MTT法检测细胞增殖情况;酶联免疫吸附测定试验（ELISA）法检测HFLS-RA中肿瘤坏死因子（TNF）-α、IL-6水平。采用免疫荧光法观察空白对照组、10 μg/L IL-1β组及10 μmol/L丹皮酚组HFLS-RA中核因子（NF）-κB p65细胞核移位情况;Western blot法检测空白对照组,10 μg/L IL-1β组,0.1、1、10 μmol/L丹皮酚组及阴性对照组（仅10 μmol/L丹皮酚处理）的基质金属蛋白酶（MMP）-1、MMP-3和Toll样受体-4（TLR4）蛋白表达。将24只雄性DBA/1小鼠随机均分为对照组、CIA组和治疗组,对小鼠进行四肢累计评分,HE染色观察关节组织病理学变化,ELISA法检测滑膜组织中TNF-α、IL-1β水平。结果　丹皮酚对IL-1β诱导HFLS-RA细胞增殖无明显抑制作用。与空白对照组相比,10 μg/L IL-1β组TNF-α、IL-6、NF-κB p65核移位、MMP-1、MMP-3和TLR4蛋白表达水平均升高,而经丹皮酚干预后可逆转这些改变（P＜0.05）。与CIA组相比,治疗组小鼠四肢累计评分明显下降（P＜0.05）;HE染色结果示丹皮酚治疗48 d时踝关节病理变化有所改善,其中炎性细胞明显下降,软骨破坏减轻。治疗组小鼠滑膜组织中TNF-α、IL-1β水平低于CIA组。结论　丹皮酚具有减轻IL-1β诱导HFLS-RA炎性因子表达和缓解CIA小鼠关节炎症状的作用,其机制可能与抑制TLR-4-NF-κB p65通路,减轻炎症反应有关。     

丹皮酚影响依托泊苷对人胶质母细胞瘤细胞U251的细胞毒性

目的:研究丹皮酚(Paeonol,Pae)对人胶质母细胞瘤细胞增殖的影响及其对化疗药物敏感性的影响。方法:应用水溶性四唑盐法,检测不同浓度和时间点Pae对体外培养人胶质母细胞瘤U251细胞增殖的影响,联合Pae及不同浓度依托泊苷(VP-16),检测Pae对VP-16细胞毒性的影响并以SPSS17.0系统软件计算IC50值。结果:(1)Pae可呈剂量依赖性抑制人胶质母细胞瘤U251细胞的增殖,Pae在浓度为15.63 mg.L-1即可将U251细胞存活率显著降低至82.88%±1.24%(P<0.01),IC50为203.81 mg.L-1。(2)Pae对U251细胞增殖的抑制作用随时间延长而增强。(3)Pae与VP-16联合使用,可显著增强VP-16对U251的细胞毒性,当联用Pae剂量为15.63 mg.L-1时,VP-16的IC50值降低至376.81μmol.L-1,与单独使用时的IC50值(767.34μmol.L-1)相比降低幅度达50.89%。(4)Pae增强VP-16对U251细胞毒性的作用具有时间依赖性,随给药时间延长而逐渐增强。结论:Pae可抑制人胶质母细胞瘤U251细胞增殖。Pae与化疗药物VP-16联用时可以显著增强此化疗药物对胶质母细胞瘤的细胞毒性,提高胶质母细胞瘤细胞对化疗药物的敏感性。     

高效液相色谱法测定当归养血丸中丹皮酚的含量

目的建立了高效液相色谱法测定当归养血丸中丹皮酚的检测方法。方法色谱条件为C18色谱柱;柱温为室温;流动相:甲醇-水(70∶30);流速1ml/min;检测波长:274nm。结果丹皮酚在0.0102～0.0510mg/ml范围内具有良好的线性关系(r=0.9999)。加样回收率为99.12%(RSD为2.88%,n=9)。结论比种方法简便,准确度高。