Development and validation of a simple and sensitive liquid chromatography–tandem mass spectrometry method for quantifying trimetazidine in human plasma

1. A simple and sensitive liquid chromatography–tandem mass spectrometry (LC‐MS‐MS) method for quantifying trimetazidine in human plasma was developed and validated. Sample preparation was based on deproteinating with acetonitrile. 2. Chromatography was performed on a C18 analytical column (5 μm; 150 × 2.1 mm i.d.) and the retention times for trimetazidine and cetirizine (used as the internal standard) were 1.8 and 3.0 min, respectively. The ionization was optimized using an electrospray ionization source and enhanced selectivity was achieved using tandem mass spectrometry. The calibration curve ranged from 0.1 to 200 ng/mL. The inter‐day precision, accuracy and the relative standard deviation (RSD) were all < 15%. The analyte was shown to be stable over the time‐scale of the entire procedure. 3. The robustness of the method was demonstrated by the good reproducibility of the results obtained during the analysis of clinical samples.     

Pharmacological strategies against cold ischemia reperfusion injury

Importance of the field: Good organ preservation is a determinant of graft outcome after revascularization. The necessity of increasing the quality of organ preservation, as well as of extending cold storage time, has made it necessary to consider the use of pharmacological additives.

Areas covered in this review: The complex physiopathology of cold-ischemia–reperfusion (I/R) injury – and in particular cell death, mitochondrial injury and endoplasmic reticulum stress – are reviewed. Basic principles of the formulation of the different preservation solutions are discussed.

What the reader will gain: Current strategies and new trends in static organ preservation using additives such as trimetazidine, polyethylene glycols, melatonin, trophic factors and endothelin antagonists in solution are presented and discussed. The benefits and mechanisms responsible for enhancing organ protection against I/R injury are also discussed. Graft preservation was substantially improved when additives were added to the preservation solutions.

Take home message: Enrichment of preservation solutions by additives is clinically useful only for short periods. For longer periods of cold ischemia, the use of such additives becomes insufficient because graft function deteriorates as a result of ischemia. In such conditions, the preservation strategy should be changed by the use of machine perfusion in normothermic conditions.     

丹红注射液联合西药治疗不稳定型心绞痛疗效分析

目的观察丹红注射液联合西药治疗不稳定型心绞痛的临床疗效。方法将100例不稳定型心绞痛患者按随机数字表法分为治疗组与对照组,每组50例。对照组予曲美他嗪及硝酸酯类制剂、β受体阻滞剂、调脂等常规西药治疗,治疗组在对照组基础上加用丹红注射液。两组疗程均为2周,观察临床疗效,并比较治疗期间心血管事件发生率及不良反应发生情况。结果①治疗组、对照组总有效率分别为98.00%和82.00%;组间临床疗效比较,差异有统计学意义(P0.05)。②治疗期间,治疗组、对照组心血管事件发生率分别为2.00%和18.00%,差异有统计学意义(P0.05)。③治疗期间,治疗组、对照组不良反应发生率分别为10.0%与8.0%,差异无统计学意义(P0.05)。结论丹红注射液联合西药治疗不稳定型心绞痛,临床疗效较好,心血管事件发生率较低。     

环磷腺苷葡胺联合曲美他嗪治疗病毒性心肌炎70例临床观察

目的:探讨环磷腺苷葡胺联合曲美他嗪治疗病毒性心肌炎患者的临床疗效。方法:将病毒性心肌炎140例患者随机分成治疗组和对照组,每组70例。两组均常规给予黄芪注射液、大剂量维生素C、辅酶A、三磷酸腺苷治疗。治疗组在常规治疗的基础上使用环磷腺苷葡胺180mg加入5%葡萄糖注射液250ml静脉滴注,每日一次,口服曲美他嗪片20mg/次,3次/天,治疗2周;对照组在常规治疗的基础上口服曲美他嗪片20mg/次,3次/天,治疗2周。结果:治疗组有效率为91.43%,对照组有效率为70%,两组间比较有统计学差异(P0.05),治疗组患者症状改善程度、心电图及心肌酶学恢复优于对照组(P0.05),不良反应无明显差异。结论:环磷腺苷葡胺联合曲美他嗪治疗病毒性心肌炎患者临床疗效好,值得推广应用。     

HPLC法测定盐酸曲美他嗪缓释片中有关物质

目的建立盐酸曲美他嗪缓释片中有关物质的高效液相色谱测定方法。方法采用Diamonsil C18色谱柱（250mm×4.6 mm,5μm）,流动相为0.012 mmol/L庚烷磺酸钠溶液（p H 3.5）–甲醇–乙腈（55∶36∶9）,体积流量为1.0 m L/min,柱温为30℃,检测波长为210 nm,进样量：20μL。结果在所选定的液相色谱条件下,有关物质与主药分离良好。盐酸曲美他嗪、杂质A、杂质B和杂质C的检出限分别为10.2、4.4、4.5、8.0 ng。结论该方法操作简单,重复性好,专属性强,可用于控制盐酸曲美他嗪缓释片中的有关物质。     

曲美他嗪对老老年慢性心力衰竭患者自主神经功能干预的配对交叉研究

目的研究曲关他嗪对常规治疗的老老年（年龄〉80岁）慢性心力衰竭患者心率变异性的影响,用以明确曲美他嗪在老老年心功能较差的患者是否能改善受损的自主神经功能。方法选择慢性心力衰竭老老年患者20对,其中男性12对,女性8对。纳入标准依据Framingham心力衰竭诊断标准,剔除含有中途退出试验的对子。病例采用两处理二阶段配对交叉设计,将符合纳入标准的病例按配对条件进行配对,再用随机分配的方法将其中之一（A组）先采用A处理方式（服用曲美他嗪20mg,每日3次）,再用B处理方式（不服用曲美他嗪）,另一研究对象（B组）则先用B处理方式再用A处理方式,使这两种处理方式在研究过程中交叉进行,患者单盲。分别在试验开始前、第2周末、第5周末进行动态心电图检查,结束试验。观察指标为心率变异性参数：24小时全部正常窦性R—R间期标准差（SDNN）;24小时内每5分钟时间段窦性R-R间期平均值的标准差（SDANN）;24小时全部正常窦性R—R间期差值的均方根（rMSSD）;心率变异三角指数（HRVTI）。结果A组的心率变异性增加具有统计学意义（FSDNN=226．024,FSDANN=57．654,FrMSSD=2735．080,FHRVTI=85．462）（P〈0．05或〈0．01）。结论除常规治疗外,应用曲美他嗪治疗老老年慢性心力衰竭,能显著提高患者的心率变异性,改善自主神经功能,具有不依赖于其他改善血流动力学药物的显著正性作用,改善心肌细胞代谢可能是此方面作用的主要基础。     

曲美他嗪对冠心病合并左心功能不全的效果

目的 探讨冠心病合并左心功能不全患者行曲美他嗪治疗的临床效果.方法 选择医院于2016年3月—2017年4月期间接收的老年冠心病合并左心功能不全患者60例,采用随机数字表法形式均分为研究组和对照组,每组均30例.对照组行基础治疗,研究组在此基础上口服曲美他嗪,记录两组心功能指标以及不良反应发生率.结果 研究组治疗后LVEDD、LVESD指标低于对照组,左心室射血分数高于对照组(P<0.05);研究组和对照组在用药过程中均未出现严重的反应,研究组出现3例转氨酶上升,2例胃肠道不适;对照组出现2例转氨酶上升,组间对比P<0.05.结论 曲美他嗪行冠心病合并左心功能不全治疗效果明显,心肌功能以及血脂水平有所改善.     

针刺联合曲美他嗪治疗心力衰竭临床评价

目的探讨针刺联合曲美他嗪治疗心力衰竭(简称心衰)的临床疗效。方法选取医院2017年4月至2019年4月收治的心衰患者86例,随机分为观察组和对照组,各43例。两组患者均予心衰常规治疗及口服盐酸曲美他嗪片,观察组患者联用针刺治疗。两组均持续治疗2周。结果观察组总有效率为88.37%,显著高于对照组的69.77%(P<0.05);治疗后,观察组患者的左室收缩末期内径(LVESd)、左室舒张末期内径(LVEDd)、QT间期离散度(QTd)及心肌肌钙蛋白I(c Tn I)、肌酸激酶同工酶(CK-MB)、肌红蛋白(MYO)水平均显著低于对照组,左室射血分数(LVEF)显著大于对照组(P<0.05),生理健康、心理健康、社会关系和周围环境评分均显著高于对照组(P<0.05);观察组与对照组不良反应发生率相当(16.28%比9.30%,χ2=0.938,P>0.05)。结论针刺联合曲美他嗪治疗心衰,能改善心功能,降低血清学指标水平,显著改善患者的生活质量。     

Effects of trimetazidine on the calcium transport and oxidative phosphorylation of isolated rat heart mitochondrial

Trimetazidine (TMZ) added in vitro to isolated cardiac mitochondria at concentrations 10–100 M in the presence of 25–100 nM extramitochondrial Ca²⁺ increased Ca²⁺ uptake and matrix Ca²⁺ concentration. This effect was less evident in the presence of physiologically Na⁺ and Mg²⁺ extramitochondrial concentrations since only 100 M TMZ was able to increase mitochondrial Ca²⁺ entry in the presence of 100 nM Ca²⁺. The drug stimulated a Ca²⁺-cooperative effect on mitochondrial Ca²⁺ transport but did not modify the rate of Ca²⁺ egress stimulated by 10 mM NaCl. An increase in mitochondrial Ca²⁺ level produced by TMZ enhanced oxoglutarate dehydrogenase activity and then ATP synthesis, particularly when 50 nM extramitochondrial Ca²⁺ was used. These data suggest that a possible cardiac mechanism of action of TMZ at mitochondrial level could support ATP synthesis by elevating the mitochondrial Ca²⁺ level.     

Effects of trimetazidine in ethanol- and acetic acid-induced colitis: oxidant/anti-oxidant status

There is overwhelming evidence in favour of a significant role of reactive oxygen metabolites (ROM) in the pathophysiology of inflammatory bowel disease (IBD) in man and in experimental animal models. This study was undertaken to investigate the possible protective effects of pretreatment with trimetazidine (TMZ) on the oxidant–anti-oxidant balance in ethanol- and acetic acid-induced colonic damage in rats. TMZ was chosen because of its various cytoprotective features (preserving cellular ATP levels, limiting intracellular acidosis and limiting inorganic phosphate, Na⁺ and Ca²⁺ accumulation) and anti-oxy characteristics which were previously reported. A total of 80 rats were randomized into eight major groups each consisting of 10 animals. Animals in groups 1, 2 and 3 served as models of ethanol-induced colitis (0.25 ml of 30% (v/v) ethanol), while group 4 served as their control. Animals in groups 5, 6 and 7 served as models of acetic acid-induced colitis (1 ml of 4% (v/v) acetic acid), while group 8 served as their control. TMZ was administered 5 mg/kg by intrarectal (i.r.) and intraperitoneal (i.p.) routes to groups 1, 2, 5 and 6. Intraperitoneal administration of TMZ was used in order to evaluate its systemic effect while i.r. administration was used to determine its local effect. After decapitation, colon mucosa samples were obtained and evaluated macroscopically and microscopically. Myeloperoxidase (MPO) activities as markers for inflammation, malondialdehyde (MDA) levels as markers for oxidant stress and reduced glutathione (GSH) and oxidized glutathione (GSSG) levels as markers for anti-oxidant status were determined. Acute colitis was observed in macroscopic and microscopic evaluation in ethanol- and acetic acid-administered groups compared with controls (P = 0.000). The macroscopic and microscopic scores in colitis groups were correlated with MPO activities (r = 0.5365, P = 0.000 and r = 0.5499, P = 0.000, respectively). MDA and GSSG levels in the acetic acid-induced colitis group were higher compared with ethanol-induced colitis group (P < 0.008 and P < 0.005, respectively), while GSH levels were significantly lower (P < 0.05). While TMZ pretreatment did not improve the oxidant state, it preserved the GSH levels significantly (P < 0.05). In conclusion, ethanol- and acetic acid-induced colitis models are appropriate experimental colitis models which in many ways manifest the characteristics seen in tissue injury related to colitis in humans. Of these two, the acetic acid-induced colitis model proved more suitable than the ethanol model for investigating the alterations in long-term and in more severe tissue injury. While TMZ pretreatment via i.p. or i.r. route did not improve the oxidative-inflammative state in either of these models, it did contribute significantly to the preservation of the anti-oxidant pool via the conservation of intracellular GSH levels. This conserving effect of TMZ was substantially more pronounced in the i.p. route compared with the i.r. route. Based on our results, we conclude that the ‘GSH-preservation’ role of TMZ can be the mode of action it manifests as an anti-oxy compound.