芪参益气滴丸对心脏后负荷增加大鼠不同阶段心脏全基因表达的影响

目的探讨芪参益气滴丸对后负荷增加大鼠不同时间点心脏全基因谱表达的影响。方法纳入雄性SD大鼠110只,术前随机取10只进行处理,相关参数作为基线数据;其余100只,随机分为假手术组(2周亚组、8周亚组、18周亚组)、模型组(2周亚组、8周亚组、18周亚组),芪参益气滴丸低剂量组(8周亚组、18周亚组)、芪参益气滴丸高剂量组(8周亚组、18周亚组),每个亚组10只。模型组大鼠采用缩窄腹主动脉的方法建立心脏后负荷增加模型,假手术组大鼠腹主动脉只穿线不进行缩窄。芪参益气滴丸组手术同模型组,术后2周开始每日给予芪参益气滴丸流浸膏灌胃处理(低剂量组150 mg/kg·d,高剂量组300 mg/kg·d)。各组大鼠分组后分别在术前、术后2周、8周、18周采用小动物超声实时影像系统及颈动脉插管检测大鼠心脏结构及血流动力学相关指标。完成上述步骤后,处死大鼠,取左心室,进行大鼠心脏RNA建库,使用Illumina HiSeqTM2500/MiseqTM对文库进行高通量测序,并进行生物信息分析。结果造模术后第8周、第18周,芪参益气滴丸组与模型组比较,能够改善后负荷增加大鼠心脏每搏输出量、左室射血分数、左室内压最大上升速率,减小平均动脉压、左室舒张末内径、左室舒张末压。芪参益气滴丸低剂量组、高剂量组大鼠在造模术后第8周与基线时全基因表达差异表达基因的个数增高,分别为2050、1710个基因,至第18周差异表达的基因个数回落,分别为1741、1558个基因。芪参益气滴丸低剂量组大鼠在第8周、第18周较模型组2周亚组差异表达的基因个数呈降低趋势,分别为2011、1557个基因;芪参益气滴丸高剂量组大鼠在第8周、第18周较模型组2周亚组差异表达的基因个数呈升高趋势,分别为1183、1554个基因。术后8周,芪参益气滴丸低剂量组、高剂量组大鼠,较模型组8周亚组差异表达的基因个数分别为46、78个基因;术后18周,芪参益气滴丸低剂量组、高剂量组大鼠,较模型组18周亚组差异表达的基因个数分别为52、32个基因。GO富集柱状图显示,造模术后第8周,芪参益气滴丸高剂量组与模型组大鼠差异基因主要集中于生物过程中的离子转运、化学刺激反应、金属离子运输以及分子功能中水解酶活力、结合功能等;造模术后第18周,芪参益气滴丸高剂量组与模型组大鼠差异基因相对分散,主要集中于分子功能中的催化活性、肽酶活性、细胞骨架蛋白结合等。KEGG富集分析结果显示,造模术后第8周,芪参益气滴丸高剂量组与模型组大鼠差异基因主要富集于昼夜节律、甘油磷脂代谢等通路中;造模术后第18周,芪参益气滴丸高剂量组与模型组大鼠差异基因主要主要富集于HIF-1信号通路。结论芪参益气滴丸可以改善后负荷增加大鼠心脏血流动力学及心脏重构,可能与药物引起造模大鼠心脏基因表达谱的显著变化,导致心肌细胞及基质细胞等生物过程、细胞结构、分子功能等一系列信号通路的变化有关。

Effect of Qishenyiqi dropping pill on the expression of cardiac whole gene in rats with increased cardiac afterload at different stages

Objective To investigate the effect of Qishenyiqi dropping pill on the expression of cardiac whole gene spectrum at different time points in rats with increased afterload.Method 110 male SD rats were included,10 of them were randomly selected for treatment before operation,and relevant parameters were taken as baseline data;the rest 100 rats were randomly divided into sham operation group(2-week sub group,8-week sub group,18 week sub group),model group(2-week sub group,8-week sub group,18 week sub group),Qishenyiqi dropping pill low dose group(8-week sub group,18 week sub group),Qishenyiqi dropping pill high dose group(8-week sub group,18 week sub group)and high dose group(8-week sub group,18 week sub group),10 in each subgroup.The rats in the model group were used to narrow the abdominal aorta to establish the model of increased cardiac afterload,while the rats in the sham operation group were only threaded without narrowing the abdominal aorta.The operation of Qishenyiqi dropping pill group was the same as that of the model group.Two weeks after the operation,Qishenyiqi dropping pill was administrated daily by gavage with liquid extract(150 mg/kg·d in low dose group and 300 mg/kg·d in high dose group).The real-time ultrasound imaging system and carotid intubation were used to detect the cardiac structure and hemodynamic indexes before the operation,2 weeks,8 weeks and 18 weeks after the operation.After completing the above steps,the rats were killed,the left ventricle was taken,the rat heart RNA library was established,and the library was sequenced by Illumina hiseqTM2500/miseqTM with high-throughput,and the biological information was analyzed.Results Compared with the model group,Qishenyiqi dropping pill group could increase the cardiac output,left ventricular ejection fraction,the maximum rate of left ventricular pressure rise,decrease the mean arterial pressure,left ventricular end diastolic diameter and left ventricular end diastolic pressure.The number of differentially expressed genes in the low dose group and high dose group of Qishenyiqi dropping pill increased from the 8th week to the baseline,which was 2050 and 1710 genes respectively.The number of differentially expressed genes decreased from the 18th week,which was 1741 and 1558 genes respectively.At the 8th and 18th week,the number of differentially expressed genes in the low dose group was lower than that in the model group at the 2nd week,which was 2011 and 1557 genes respectively;at the 8th and 18th week,the number of differentially expressed genes in the high dose group was higher than that in the model group at the 2nd week,which was 1183 and 1554 genes respectively.At 8 weeks after operation,the number of differentially expressed genes in the low-dose group and high-dose group of Qishenyiqi dropping pill was 46 and 78 genes respectively,compared with the model group at 8 weeks;at 18 weeks after operation,the number of differentially expressed genes in the low-dose group and high-dose group of Qishenyiqi dropping pill was 52 and 32 genes respectively,compared with the model group at 18 weeks.The histogram of GO enrichment showed that at the 8th week after modelling,the different genes of high dose group and model group were mainly concentrated in the ion transport,chemical stimulation,metal ion transport,hydrolase activity and binding function in biological process;at the 18th week after modelling,the different genes of high dose group and model group were relatively dispersed It mainly focuses on the catalytic activity,peptidase activity,cytoskeleton protein binding and so on.The results of KEGG enrichment analysis showed that at the 8th week after the model making operation,the different genes in the high dose group and the model group were mainly enriched in circadian rhythm,glycerophospholipin metabolism and other pathways;at the 18th week after the model making operation,the different genes in the high dose group and the model group were mainly enriched in HIF-1 signalling pathway.Conclusion Qishenyiqi dropping pill can improve the cardiac hemodynamics and cardiac remodelling in rats with increased afterload,which may be related to the significant changes of gene expression in the heart of model rats caused by drugs,and the changes of a series of signal pathways such as biological process,cell structure and molecular function of myocardial cells and stromal cells.