贵州医科大学学报

2016, v.41;No.194(11) 1301-1306

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剪刀股总酚酸提取条件的优化及对小鼠缺血心肌的保护作用
Optimization of Extraction of Total Phenolic Acid from Ixeris Debilis A. Gray and Its Protective Effect on Myocardial Ischemia in Mice

王威;许丽娟;
WANG Wei;XU Lijuan;The 3~(th) Hospital of Zhangjiakou;The 4~(th) Hospital of Zhangjiakou;

摘要(Abstract):

目的:优化剪刀股总酚酸的提取方法,观察剪刀股总酚酸对小鼠缺血心肌的保护作用。方法:通过正交试验优化剪刀股总酚酸的提取条件,并采用高效液相色谱法测定没食子酸、原儿茶酸、绿原酸的含量;选取昆明种小鼠60只,随机分为正常对照组,模型对照组,阳性对照组,剪刀股总酚酸高、中、低剂量给药组共6组,采用异丙肾上腺素法制备心肌缺血小鼠模型,测定血清中乳酸脱氢酶(LDH)、肌酸激酶MB(CK-MB)的活性及心肌中超氧化物歧化酶(SOD)的活性和丙二醛(MDA)的含量,并通过Nagar-Olsen染色观察小鼠心肌缺血损伤程度。结果:剪刀股总酚酸最佳提取条件为提取温度为70℃,液料比为10∶1,醇浓度为15%,提取90 min(30 min×3);剪刀股总酚酸平均提取率为4.85%,RSD为1.988%;剪刀股中没食子酸、原儿茶酸和绿原酸分别在0.60~9.60、0.90~14.40、1.00~16.00 mg/L,与峰面积呈良好线性关系,平均回收率分别为97.27%、98.15%、97.17%(n=9);与模型组相比,阳性对照组和剪刀股总酚酸(高、中剂量)组心肌缺血小鼠血清中LDH、CK-MB活性显著降低,心肌中SOD的活性升高,心肌MDA的含量降低(P<0.05);Nagar-Olsen染色显示,高、中剂量的剪刀股总酚酸组及阳性对照组小鼠心肌缺血损伤红染面积明显减少。结论:剪刀股中总酚酸含量较高,剪刀股总酚酸对心肌缺血小鼠的心肌有一定保护作用。
Objective: To optimize the extraction of total phenolic acid from Ixeris debilis A. Gray and to explore its protective effect on myocardial ischemia induced by isoproterenol in mice. Method: The total phenolic acid extraction conditions from Ixeris debilis A. Gray were optimized by orthogonal test,and HPLC method was adopted for the determination of gallic acid,protocatechuic acid,chlorogenic acid content. 60 Kunming mice were divided into 6 groups: normal control group,model control group,positive control group,low dose group,middle dose group and high dose groups. A mouse model of myocardial ischemia was prepared by the method of isoproterenol. The effect of total phenolic acid in Ixeris debilis A. gray on myocardial ischemia model was studied by measuring the activities of serum lactate dehydrogenase(LDH),serum creatine kinase MB(CK-MB),the activities of the superoxide dismutase(SOD) and the content of malondialdehyde(MDA) in myocardium. The degree of myocardial ischemic injury in mice was observed by Nagar-Olsen staining. Result: The best condition forextracting the total phenolicacid from Ixeris debilis A. Gray was heating the Ixeris debilis A. Gray in 10times( Volume / quality) alcohol( 15%) at 70℃ for 90( 30 × 3) minutes. The average extraction yield of the total phenolic acid in Ixeris debilis A. Gray was 4. 85%( RSD = 1. 988%). The contents of gallic acid,protocatechuic acid and chlorogenic acid in Ixeris Debilis A. Gray showed good linear relationships with the peak area in the ranges of 0. 60 ~ 9. 60,0. 90 ~ 14. 40 and 1. 00 ~ 16. 00 μg / mL,respectively,and the average recoveries( n = 9) were 97. 27%,98. 15%,and 97. 17%,respectively.Compared with the model group,the activities of LDH,CK-MB in serum and the content of myocardial MDA of mice in the positive control group and the total phenolic acid groups( high,middle dose) were decreased and the activities of myocardial SOD were increased. The results of Nagar-Olsen staining showed that the area of cardiac muscles injured by ischemia in the positive control group and the total phenolic acid groups( high,middle dose) were diminished significantly. Conclusion: The content of total phenolic acid in Ixeris debilis A. Gray is at a relatively high level,which has a protective effect on the myocardium of mice with myocardial ischemia.

关键词(KeyWords): 心肌缺血;异丙肾上腺素;小鼠;剪刀股;总酚酸
myocardial ischemic;isoproterenol;mice;Ixeris debilis A. Gray;total phenolic acid

Abstract:

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作者(Author): 王威;许丽娟;
WANG Wei;XU Lijuan;The 3~(th) Hospital of Zhangjiakou;The 4~(th) Hospital of Zhangjiakou;

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DOI: 10.19367/j.cnki.1000-2707.2016.11.014

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