高效液相色谱法测定人肝细胞色素P450 3A4转基因细胞中氨氯地平

目的 :研究氨氯地平经人肝细胞色素 P4 5 0 3A4 (CYP3A4 )的代谢 ,建立反相高效液相色谱的测定人肝转基因细胞 CYP3A4酶 S9孵育液中氨氯地平浓度的方法。方法 :与人肝转基因细胞提取的 S9上清液孵育之后的样品 ,用 3倍量的甲醇沉淀后离心 ,取上清液用 0 .4 5μm微孔滤膜滤过后进样。采用 Hypersil C18柱 ,以乙腈 -磷酸盐缓冲液 (4 5∶ 5 5 ,v/ v,p H4 .5 )为流动相 ,普萘洛尔为内标 ,在 2 5 0 nm波长处测定。结果 :氨氯地平浓度在 0 .2～ 30 .0μg/ ml范围内线性关系良好 ,r=0 .9993,方法平均回收率为 (98.2± 2 .4 ) % (n=5 ) ,日内和日间相对标准偏差(RSD)均小于 10 % ,检测限为 2 0 ng/ ml,定量限为 0 .2 μg/ ml(回收率为 10 4 .0 % ,RSD为 11.4 % ,n=5 )。结论 :建立的反相高效液相色谱方法简便、准确 ,可用于研究氨氯地平在人肝 CYP3A4转基因细胞中的代谢。     

Effect of nilvadipine on the voltage-dependent Ca channels in rat hippocampal CA1 pyramidal neurons

Effects of nilvadipine on the low- and high-voltage activated Ca²⁺ currents (LVA and HVA I_Ca, respectively) were compared with other organic Ca²⁺ antagonists in acutely dissociated rat hippocampal CA1 pyramidal neurons. The inhibitory effects of nilvadipine, amlodipine and flunarizine on LVA I_Ca were concentration- and use-dependent. The apparent half-maximum inhibitory concentrations (IC₅₀s) at every 1- and 30-s stimulation were 6.3×10⁻⁷ M and 1.8×10⁻⁶ M for flunarizine, 1.9×10⁻⁶ M and 7.6×10⁻⁶ M for nilvadipine, and 4.0×10⁻⁶ M and 8.0×10⁻⁶ M for amlodipine, respectively. Thus, the strength of the use-dependence was in the sequence of nilvadipine>flunarizine>amlodipine. Nilvadipine also inhibited the HVA I_Ca in a concentration-dependent manner with an IC₅₀ of 1.5×10⁻⁷ M. The hippocampal CA1 neurons were observed to have five pharmacologically distinct HVA Ca²⁺ channel subtypes consisting of L-, N-, P-, Q- and R-types. Nilvadipine selectively inhibited the L-type Ca²⁺ channel current which comprised 34% of the total HVA I_Ca. On the other hand, amlodipine non-selectively inhibited the HVA Ca²⁺ channel subtypes. These results suggest that the inhibitory effect of nilvadipine on the neuronal Ca²⁺ influx through both LVA and HVA L-type Ca²⁺ channels, in combination with the cerebral vasodilatory action, may prevent neuronal damage during ischemia.     

氨氯地平抑制氧化型胆固醇诱导的血管内皮细胞凋亡

目的:研究氧化型胆固醇(Triol与25-OH)对人脐静脉内皮细胞凋亡的诱导作用,并观察氨氯地平对氧化型胆固醇诱导内皮细胞凋亡的影响。方法:体外培养人脐静脉内皮细胞,利用光镜、电镜、DNA电泳及流式细胞仪测定作为凋亡检测指标。结果:对照及胆固醇组未检测到细胞凋亡,Triol与25-OH处理组光镜、电镜下可见典型的细胞凋亡形态学改变,电泳示"DNAladder",流式细胞仪检测出现亚二倍体峰,凋亡率分别为32.25%与23.04%,而氨氯地平使其凋亡率分别下降至13.42%与11.77%。结论:氧化型胆固醇(Triol与25-OH)可以诱导人脐静脉内皮细胞发生凋亡,而氨氯地平抑制其凋亡诱导作用。     

氨氯地平与左旋氨氯地平治疗夜间高血压疗效比较

目的 :观察左旋氨氯地平与氨氯地平治疗夜间高血压患者的疗效及其对血压昼夜节律的影响。　方法 :6 0例轻、中度原发性高血压患者 (必须同时具备夜间血压增高 )随机接受氨氯地平 5mg或左旋氨氯地平 2 .5mg(各30例 )治疗 4周。用 2 4h动态血压监测治疗前后血压的昼夜节律变化。　结果 :氨氯地平和左旋氨氯地平降压疗效显著 ,但二药之间比较无显著差异 (P >0 .0 5 ) ,均不影响血压昼夜节律。　结论 :氨氯地平和左旋氨氯地平治疗有夜间血压增高的轻、中度高血压患者疗效好 ,而且安全     

Unaltered insulin sensitivity during calcium channel blockade with amlodipine

Summary The sensitivity of peripheral tissues to insulin is of pathophysiological, therapeutic and possibly also of prognostic relevance. Calcium channel blockers are widely used in the treatment of cardiovascular disorders that are commonly associated with decreased insulin sensitivity (S_I). To evaluate the effects of calcium channel blokkade on S_I, glucose homoeostasis and lipid profiles, studies were made of S_I (determined by the Minimal Model Method of Bergman), basal glucose and insulin levels, serum total triglyceride (Tg) and lipoprotein cholesterol (C) fractions and certain other variables in 38 healthy young men (24 y) during placebo and after 3 weeks of calcium channel blockade with amlodipine 5 mg once daily. Measurements were made after 3 days on a standard diet (2200 kcal · day^–1, 45% carbohydrates, 40% fat and 15% proteins) and after an overnight fast. Compared to placebo, amlodipine decreased supine systolic blood pressure (P<0.01). Heart rate, body weight and 24 h urinary sodium excretion were unaltered, and so were fasting plasma glucose (placebo vs amlodipine: 4.86 vs 4.83 mmol·1^–1, respectively) and insulin levels (7.7 vs 7.9 U·ml^–1), S_I (10.5 vs 9.6·10^–4 × min^–1 pro U·ml^–1), serum total Tg, C and lipoprotein C fractions.The findings demonstrate unchanged insulin sensitivity and secretion, as well as lipoprotein regulation, during maintenance administration of 5 mg amlodipine daily to healthy young men.This work was supported in part by the Swiss National Science Foundation     

Simultaneous determination of amlodipine,valsartan and hydrochlorothiazide by LC–ESI-MS/MS and its application to pharmacokinetics in rats

Polypill is a fixed-dose combination that contains three or more active ingredients used as a single daily pill to achieve a large effect in preventing cardiovascular disease with minimal adverse effects. A novel and accurate liquid chromatography tandem mass spectrometry method using electrospray ionization mode has been developed and validated for the simultaneous determination of amlodipine (AMD), valsartan (VAL) using losartan (LOS) as an internal standard (IS), and hydrochlorothiazide (HCT) using furosemide (FSD) as an IS. The separation was carried on Aquasil C₁₈ (50 mm×2.1 mm, 5 µm) reversed phase column using acetonitrile and water containing 0.1% formic acid (50:50, v/v) as the mobile phase. The method was validated in terms of linearity, accuracy and precision over the concentration range of 1–1000 ng/mL. The intra and inter-day precision and accuracy, stability and extraction recoveries of all the analytes were in the acceptable range. This method can be successfully applied to the pharmacokinetic study of AMD, VAL and HCT when given as a polypill.     

氨氯地平阿托伐他汀钙片应用于高血压合并高血脂的价值

目的：探讨氨氯地平阿托伐他汀钙片治疗高血压合并高血脂的价值。方法将86例高血压合并高血脂患者随机均分为观察组与对照组,观察组给予氨氯地平阿托伐他汀钙片口服治疗,对照组给予氨氯地平口服治疗,12周后评定两组的疗效。结果治疗后,观察组患者SBP、DBP明显低于对照组(P<0.05);观察组患者TG、TC、LDL、HDL均明显优于对照组(P<0.05);观察组患者的治疗总有效率95.35%显著高于对照组的74.42%(P<0.05)。结论氨氯地平阿托伐他汀钙片治疗高血压合并高血脂疗效肯定,安全性好。     

探讨氨氯地平联合吲达帕胺对高血压合并冠心病血脂、血压的影响

目的：研究氨氯地平联合吲达帕胺治疗高血压合并冠心病的临床疗效及其对患者血脂、血压指标的影响。方法以随机数表法将该院160例高血压合并冠心病患者分为观察组与对照组,两组均为80例,对照组给予单一吲达帕胺治疗,观察组采取氨氯地平联合吲达帕胺治疗,比较两组血脂、血压指标改变,并对临床疗效及安全性进行分析。结果①治疗后观察组SBP、DBP、TC、TG、LDL为（117.87±10.67）mmHg、（81.67±6.95）mmHg、（4.36±0.68）mol/L、（1.64±0.58）mol/L、（3.84±0.47）mol/L VS 对照组（142.36±9.80）mmHg、（92.44±8.21）mmHg、（5.76±0.75）mol/L、（3.60±0.47）mol/L、（5.03±0.33）mol/L 显著较低（P＜0.05）,HDL两组比较（1.42±0.21）mol/L VS（1.16±0.17）mol/L,观察组显著较高（P＜0.05）;②观察组治疗有效率92.50% VS 对照组73.75%显著较高（P＜0.05）;③观察组总有效率96.25%VS 对照组82.50%显著较高,对比差异有统计学意义（P＜0.05）;④观察组2例头痛,1例头晕,不良反应发生率3.75%VS 对照组5.00%差异无统计学意义（P＞0.05）。结论氨氯地平联合吲达帕胺治疗高血压合并冠心病临床效果显著,可改善患者血脂及血压水平,无明显不良反应,具有较高的临床应用价值。     

高糖激活Ca~(2+)-CaN-NFAT3信号通路致H9c2细胞肥大

目的:利用细胞实验探讨不同高糖浓度培养H9c2细胞是否引起心肌细胞肥大,探讨Ca2+-Ca NNFAT3信号通路是否参与高糖引起H9c2心肌细胞肥厚过程。方法:体外培养H9c2大鼠心肌细胞48 h,分为5mmol/L糖对照组、25mmol/L糖培养组、50 mmol/L糖培养组、25mmol/L糖培养加L型钙通道阻滞剂苯磺酸氨氯地平[商品名络活喜(Norvasc)]组和50 mmol/L糖培养加Norvasc组5组。观察H9c2细胞形态并测量细胞表面积大小;荧光分光光度法检测心肌细胞内钙离子浓度[Ca2+]i;ELISA测细胞内Ca N浓度;real-time PCR法及Western blot检测Ca NAβ、NFAT3和β-MHC的mRNA及蛋白表达。结果:细胞大小、单细胞平均[Ca2+]i测定荧光值及细胞内Ca N浓度在随糖浓度升高呈阶梯上升,50 mmol/L时达到最高点。加入Norvasc后细胞大小较相同条件不加Norvasc者降低。Ca NAβ、NFAT3和β-MHC的mRNA及蛋白表达随糖浓度升高逐步升高,50 mmol/L时达最高。加入Norvasc后Ca NAβ、NFAT3和β-MHC的mRNA及蛋白表达均较未加Narvasc组明显降低。结论:高糖通过激活Ca2+-Ca N-NFAT3信号通路引起H9c2心肌细胞肥大。     

动态血压监测评价甲磺酸氨氯地平的降压疗效

目的:应用动态血压监测(ABPM)评价甲磺酸氨氯地平的降压疗效,并与苯磺酸氨氯地平进行比较。方法:77例高血压患者按不同的治疗药物随机分为甲磺酸氨氯地平组、苯磺酸氨氯地平组,均治疗12周,治疗前后测定动态血压、心率、诊室血压、生化指标。结果:治疗12周末,2组的诊室血压,24h、白昼及夜间平均血压、平均脉压、血压负荷均显著降低(P<0·05或P<0·01),心率无显著改变。12周末甲磺酸氨氯地平组的总有效率在诊室血压为92·3%,在ABPM为71·8%;苯磺酸氨氯地平组分别为92·1%和68·4%,2组间均差异无统计学意义。甲磺酸氨氯地平组收缩压和舒张压的24h降压谷/峰比分别为70%和72%;苯磺酸氨氯地平组分别为66%和69%。2组患者出现的不良反应均轻微。结论:甲磺酸氨氯地平与苯磺酸氨氯地平一样能平稳、有效、安全地降低血压,可作为一线降压用药。