2020 年第 2 期 第 15 卷

采用液滴萃取表面分析-串联质谱法检测真武汤中有效成分在心力衰竭模型大鼠体内的分布

Distribution of active constituents of Zhenwu decoction detected by liquid extraction surface analysis and tandem mass spectrometry in heart failure rats

作者：仇琪1曹景琳1郝晓艳2张芸楠1林阳1

英文作者：

单位：1首都医科大学附属北京安贞医院药事部100029；2首都医科大学附属北京安贞医院超声心动科100029

英文单位：

关键词：心力衰竭；真武汤；液滴萃取表面分析；心功能；体内分布

英文关键词：

• 摘要：

• 【摘要】目的    采用液滴萃取表面分析-串联质谱法（LESA-MS/MS）对真武汤中有效成分进行检测，初步探讨真武汤对心力衰竭模型大鼠的治疗作用及其有效成分在模型大鼠体内的分布。方法    雄性SD大鼠40只，采用冠状动脉左前降支结扎的方式制备心肌梗死后心力衰竭模型30只；另制作假手术动物10只（假手术组），只穿线不结扎。成功造模的大鼠（20只）完全随机分为模型对照组和真武汤组，每组10只。术后当天开始给药，真武汤组以羧甲基纤维素钠溶解方式灌胃给予真武汤粉剂，假手术组和模型对照组灌胃给予同体积羧甲基纤维素钠，连续给药60 d。其后对大鼠进行超声心动图检测。取大鼠心脏边缘区、肝脏和肾脏组织制备冰冻切片，通过LESA-MS/MS测定真武汤成分（茯苓酸、次乌头碱、去甲乌药碱）在不同组织中的分布。结果    给药60 d后，模型对照组和真武汤组的左心室射血分数（LVEF）和左心室短轴缩短率（FS）均明显低于假手术组［（0.28±0.13）%、（0.39±0.08）%比（0.67±0.04）%，（0.27±0.02）%、（0.36±0.07）%比（0.50±0.03）%］（均P＜0.01）；真武汤组的LVEF和FS均明显高于模型对照组（均P＜0.05）。LESA-MS/MS结果显示，与模型对照组比较，真武汤组茯苓酸、次乌头碱和去甲乌药碱在大鼠心脏边缘区、肝脏、肾脏组织中的信号强度均明显增加（均P＜0.01）；真武汤有效成分在以上组织中的信号强度均为：去甲乌药碱＞茯苓酸＞次乌头碱。结论    真武汤对心力衰竭大鼠的心功能有明显改善作用。真武汤的有效成分茯苓酸、次乌头碱和去甲乌药碱在心力衰竭大鼠的心脏边缘区、肝脏和肾脏均有分布，与中药理论中附子的归心、肾经和茯苓归心、肝经的记载相印证。心主血脉，肾主水，肝主疏泄，提示真武汤有效成分发挥作用可能与水液代谢相关。

• 【Abstract】Objective    To explore the therapeutic effect of Zhenwu decoction on heart failure rats and to detect the distribution of its active constituents in different tissues using liquid extraction surface analysis and tandem mass spectrometry(LESA-MS/MS). Methods    Forty male SD rats were used in this study. Thirty of them were treated with left anterior descending coronary artery ligation to prepare heart failure models after myocardial infarction. Ten rats were prepared as sham operation group following an identical procedure without actual tying of suture. Twenty heart failure model rats were randomly divided into model control group and Zhenwu decoction group, with 10 rats in each group. After operation, the Zhenwu decoction group had intragastric administration of Zhenwu decoction powders dissolved by carboxymethylcellulose sodium solution, while the sham operation group and model control group had intragastric administration of equal carboxymethylcellulose sodium solution. Echocardiography was performed after 60 days of medication. Heart marginal zone, liver and kidney tissues were separated for frozen section. Contents of Zhenwu decoction constituents including poria acid, hypaconitine and higenamine in different tissues were tested by LESA-MS/MS. Results    After 60 days of medication, left ventricular ejection fraction(LVEF) and fraction shortening(FS) in the model control group and Zhenwu decoction group were significantly lower than those in the sham operation group［（0.28±0.13）%,（0.39±0.08）% vs （0.67±0.04）%; （0.27±0.02）%,（0.36±0.07）% vs （0.50±0.03）%］（all P＜0.01）; LVEF and FS in the Zhenwu decoction group were significantly higher than those in the model control group（all P＜0.05）.  LESA-MS/MS results showed that signal intensities of poria acid, hypaconitine and higenamine at heart marginal zone, Liver and kidney tissues significantly increased in the Zhenwu decoction group as compared with those in the model control group（all P＜0.01）; the order of signal intensities of the components was as follows: higenamine＞poria acid＞hypaconitine. Conclusions    Zhenwu decoction can effectively improve heart function of rats with heart failure. The active constituents of Zhenwu decoction including higenamine, poria acid and hypaconitine distribute in heart marginal zone, kidney and liver, which is consist with the Meridian Tropism Theory of traditional Chinese Medicine that aconitum carmichaeli is attributed to heart and kidney, and poris cocos is attributed to heart and liver. The theory that heart controls blood and vessel, kidney controls water, liver controls conveyance and dispersion suggests that the action of Zhenwu decoction may be related with water metabolism.