冠心Ⅱ号提取工艺改进及其对急性心肌梗死大鼠心脏功能的影响

Effects of Process Improvement Guanxin Ⅱ on Cardiac Function in Rats after Acute Myocardial Infarction

目的对冠心Ⅱ号进行提取工艺改进并观察其对急性心肌梗死(AMI)大鼠心脏功能的影响。方法将冠心Ⅱ号提取工艺进行改进和优化得到精制过滤液和纯化洗脱液,并采用高效液相色谱仪进行主要成分分析。采用左冠状动脉前降支下结扎法制备AMI模型,造模成功的50只大鼠按随机数字表法分为模型组、尼可地尔片组(2.7 mg/kg体重)、复方川丹颗粒组(川丹颗粒,4.60 g生药/kg体重)、冠心Ⅱ号精制组(4.60 g生药/kg体重)、冠心Ⅱ号纯化组(4.60 g生药/kg体重)组,每组10只。另设10只穿线不结扎为假手术组。术后第2天开始灌胃给药,连续4周,假手术组、模型组灌胃等容积纯净水。采用高频超声心动图检测各项心脏形态学和功能指标。ELSIA法检测心肌钙蛋白T(cTnT)、内皮素-1(ET-1)、降钙素基因相关肽(CGRP)、肌酸激酶同工酶MB(CK-MB)、血栓素B2(TXB2)和6-酮-前列腺素F1α(6-Keto-PGF1α)水平。TTC法染色测量心肌梗死面积,HE染色观察心肌病理形态和炎症细胞浸润状态。结果丹酚酸B和芍药苷外标含量分别为川丹颗粒0.767%、0.418%,冠心Ⅱ号精制组4.247%、1.546%,冠心Ⅱ号纯化组32.758%、12.150%。与假手术组比较,模型组ET-1、TXB2升高(P<0.05,P<0.01),心脏超声学检测指标改善(P<0.05,P<0.01)。与模型组比较,冠心Ⅱ号方两组和尼可地尔片组心脏超声学检测指标改善(P<0.05,P<0.01),大鼠心重指数、梗死面积比值、cTnT、ET-1和TXB2水平亦降低(P<0.05,P<0.01);川丹颗粒组部分心脏超声学检测指标改善(P<0.05,P<0.01),大鼠心重指数、cTnT、ET-1和TXB2降低(P<0.05,P<0.01);各给药组大鼠心肌缺血区的心肌细胞炎症浸润和细胞形态不同程度改善。结论冠心Ⅱ号提取工艺改进的主要有效成分丹酚酸B和芍药苷含量有较大提高,可提高AMI后大鼠心脏的射血分数、抑制心室重构及改善心功能,部分指标优于川丹颗粒。

Objective To improve the technology of Guanxin Ⅱ prescription and observe its effect on cardiac function in rats after acute myocardial infarction(AMI). Methods The extraction process of Guanxin Ⅱ prescription was improved and optimized to obtain the filtrate and purified eluent and the main components were analyzed by high performance liquid chromatography. AMI model rats were established by ligating left anterior descending coronary artery. Totally 50 rats were randomly divided into model group,Nicorandil Tablet group(2.7 mg/kg body weight),compound Chuandan Granule group(Chuandan Granule,4.60 g crude drug/kg body weight),refined Guanxin Ⅱ group(4.60 g raw drug/kg body weight),and purified Guanxin Ⅱ group(4.60 g crude drug/kg body weight). The sham group underwent the same procedure except for LAD ligation. 10 rats in each group. Gavage was started on the 2 nd day after surgery for 4 weeks. The sham group and model group were given equal volume of purified water by gavage. The cardiac morphological and functional parameters were detected by high frequency echocardiography. Cardiac troponin T(cTnT),endothelin-1(ET-1),calcitonin gene-related peptide(CGRP),creatine kinase isoenzyme MB(CK-MB),thrombus B2(TXB2)and 6-keto-prostaglandin F1α(6-Keto-PGF1α)were detected by ELISA. The area of myocardial infarction was measured by TTC staining,and the pathological morphology and inflammatory cell infiltration were observed by HE staining. Results The external standard contents of salvianolic acid B and paeoniflorin were 0.767% and 0.418% in Chuandan Granule group,4.247% and 1.546% in refined Guanxin Ⅱ group,and 32.758% and 12.150% in purified Guanxin Ⅱ group,respectively. Compared with the sham group,cTnT,ET-1 and TXB2 increased(P <0.05,P <0.01),the cardiac ultrasound index improved(P<0.05,P<0.01). Compared with the model group,the cardiac ultrasound index improved heart weight index,the ratio of left ventricular infarct area to myocardial area,cTnT,ET-1 and TXB2 decreased(P<0.05,P<0.01)in Nicorandil Tablet group,refined Guanxin Ⅱ group and purified Guanxin Ⅱ group;partial cardiac ultrasound index improved,cardiac weight index,cTnT,ET-1 and TXB2 decreased(P<0.05)in Chuandan Granule group;each drug group improved the inflammatory infiltration and morphology of myocardial cells in the ischemic area. Conclusions The content of salvianolic acid B and paeoniflorin,the main active ingredients of Guanxin Ⅱ,was significantly increased,and it also increased the ejection fraction of rat heart after AMI,inhibited ventricular remodeling and improved cardiac function. Some of the indicators are better than those of Chuandan Granule group.