阅读说明：本技术 一种丹参红花提取物及其制备方法与应用 (Red sage root and safflower extract and preparation method and application thereof ) 是由 姜苗苗 雷鹏 樊官伟 曹远林 于 2020-05-19 设计创作，主要内容包括：本发明提供了一种丹参红花提取物及其制备方法与应用,通过进一步的提取有效筛选和保留了核心功效成分,减少了药物的副作用,通过抑制葡萄糖代谢、刺激脂肪酸代谢功能改善缺血再灌注心脏的能量代谢障碍；通过抑制血小板脱颗粒、发生聚集改善血流动力学参数,预防血栓的发生；上调抗氧化应激内源性代谢标志物牛磺酸的水平,抑制脂质过氧化,保护心肌抗氧化应激损伤；还可以通过补体系统抑制缺血再灌注损伤引起的心肌炎症发应；对减轻冠心病患者的心肌缺血再灌注损伤效果显著。(The invention provides a red sage root and safflower extract and a preparation method and application thereof, which effectively screens and retains core functional components through further extraction, reduces the side effect of the medicament, and improves the energy metabolism disorder of ischemia reperfusion heart by inhibiting glucose metabolism and stimulating fatty acid metabolism function; the occurrence of thrombus is prevented by inhibiting the degranulation and aggregation of the platelets to improve hemodynamic parameters; the level of taurine which is an endogenous metabolic marker of oxidative stress resistance is adjusted up, lipid peroxidation is inhibited, and myocardial oxidative stress damage is protected; can also inhibit myocardial inflammation caused by ischemia reperfusion injury through complement system; has obvious effect of relieving myocardial ischemia reperfusion injury of patients with coronary heart disease.)

1. A red sage root and safflower extract is characterized in that: is prepared by the following steps:

(1) passing the Saviae Miltiorrhizae radix-Carthami flos concentrated water extract through octadecyl silane bonded silica gel open type chromatographic column, respectively eluting with three gradient eluents of 0.1% formic acid water, 0.1% formic acid-water methanol 90:10 solution, and 0.1% formic acid water-methanol 5:95 solution, and washing 27, 14 and 1 column volume respectively for each gradient;

(2) collecting the first 33 column volume fractions as D1 and concentrating and drying under reduced pressure;

(3) collecting fractions of 34 th and 35 th column volumes, adopting an A-phase 0.1% formic acid water-B-phase methanol elution system, further separating by an SB-C18 high performance liquid chromatography column under the ultraviolet detection wavelength of 280nm, wherein the sample injection amount is 400 mu L, the mobile phase is set as 95% A phase and 5% B phase for washing at 0-20min, the flow rate is 10mL/min, the mobile phase with the same proportion is adopted at 21-22min, and the flow rate is 8 mL/min; collecting the flow part obtained in 0-22min as D2;

(4) mixing D1 and D2, concentrating under reduced pressure, and lyophilizing.

2. The salvia miltiorrhiza and safflower extract of claim 1, wherein: is prepared by the following steps: mixing Saviae Miltiorrhizae radix and Carthami flos, soaking in warm water, and concentrating the filtrate to obtain Saviae Miltiorrhizae radix-Carthami flos concentrated water extract.

3. The salvia miltiorrhiza and safflower extract of claim 2, wherein: is prepared by the following steps: mixing the red sage root and the safflower according to the mass ratio of 3:1, soaking twice with 10 times of water at 65 ℃ for 1 hour each time, filtering, combining the filtrates, and concentrating to 1 time of volume to obtain the red sage root-safflower concentrated water extract.

4. The salvia miltiorrhiza and safflower extract of claim 1, wherein: the main components comprise glucose, galactose, arabinose, fructose, rhamnose, rutinose, rutulose, melibiose, sucrose, trehalose, succinic acid, malonic acid, formic acid, isoleucine, leucine, valine, threonine, alanine, proline, pyroglutamic acid, trigonelline and uridine.

5. The Salvia miltiorrhiza and safflower extract of claim 4, wherein: the contents (mean ± standard deviation) of each main component are respectively:

6. a method for preparing the salvia miltiorrhiza and safflower extract as claimed in claim 1, which is characterized in that: the method comprises the following specific steps:

(1) passing the Saviae Miltiorrhizae radix-Carthami flos concentrated water extract through octadecyl silane bonded silica gel open type chromatographic column, respectively eluting with three gradient eluents of 0.1% formic acid water, 0.1% formic acid-water methanol 90:10 solution, and 0.1% formic acid water-methanol 5:95 solution, and washing 27, 14 and 1 column volume respectively for each gradient;

(2) collecting the first 33 column volume fractions as D1 and concentrating and drying under reduced pressure;

(3) collecting fractions of 34 th and 35 th column volumes, adopting an A-phase 0.1% formic acid water-B-phase methanol elution system, further separating by an SB-C18 high performance liquid chromatography column under the ultraviolet detection wavelength of 280nm, wherein the sample injection amount is 400 mu L, the mobile phase is set as 95% A phase and 5% B phase for washing at 0-20min, the flow rate is 10mL/min, the mobile phase with the same proportion is adopted at 21-22min, and the flow rate is 8 mL/min; collecting the flow part obtained in 0-22min as D2;

(4) mixing D1 and D2, concentrating under reduced pressure, and lyophilizing.

7. Use of the red sage and safflower extract of claim 1 for the preparation of a medicament for preventing and alleviating/alleviating myocardial ischemia reperfusion injury.

Technical Field

The invention relates to the technical field of traditional Chinese medicines, in particular to a red sage root and safflower extract and a preparation method and application thereof.

Background

Coronary Heart Disease (CHD) is a common cardiovascular disease, and the morbidity and mortality of the CHD tend to rise year by year, thereby seriously harming the life health of human beings. In recent years, Percutaneous Coronary Intervention (PCI) has become more important as a treatment for coronary heart disease, but myocardial ischemia-reperfusion injury (MI/RI) is likely to occur. MI/RI mechanisms include oxidative stress injury, lipid peroxidation, calcium ion overload, inflammatory response, disorders of energy metabolism, apoptosis, and the like.

The Danhong prescription is composed of two medicines of red sage root and safflower, has the efficacies of promoting blood circulation by removing blood stasis, promoting blood circulation by relaxing vein, eliminating phlegm and promoting tissue regeneration, and eliminating evil and not damaging vital qi, and can be used for treating diseases such as coronary heart disease, angina pectoris, myocardial infarction, cerebral thrombosis and the like. Clinical data show that: the Danhong prescription can obviously reduce Acute Myocardial Infarction (AMI), reduce the incidence rate of MI/RI arrhythmia, relieve the injury of oxidative stress on vascular endothelium, improve the vascular endothelium function after PCI operation, inhibit the release of inflammatory factors and stabilize atheromatous plaque.

Recent researches show that the Danhong formula contains a large amount of plant nascent chemical components, and the pharmacodynamic action of the composition of the components is not reported.

Disclosure of Invention

The invention aims to solve the technical problem of providing a red sage root and safflower extract.

The invention also aims to provide a preparation method of the red sage root and safflower extract.

The invention also aims to provide application of the red sage root and safflower extract.

The technical scheme adopted by the invention is as follows:

a red sage root and safflower extract is prepared by the following method:

(1) passing the Saviae Miltiorrhizae radix-Carthami flos concentrated water extract through octadecyl silane bonded silica gel open type chromatographic column, respectively eluting with three gradient eluents of 0.1% formic acid water, 0.1% formic acid-water methanol (90:10), and 0.1% formic acid water-methanol (5:95), and washing each gradient with 27, 14 and 1 column volume respectively;

(2) collecting the first 33 column volume fractions as D1 and concentrating and drying under reduced pressure;

(3) collecting 34 th and 35 th column volume fractions, further separating with 0.1% formic acid water (A phase) -methanol (B phase) elution system at 280nm ultraviolet detection wavelength by SB-C18 high performance liquid chromatography column with sample amount of 400 μ L, washing with 95% A phase and 5% B phase at 0-20min flow rate of 10mL/min, and using the same ratio of mobile phase at 21-22min flow rate of 8 mL/min; collecting the flow part obtained in 0-22min as D2;

(4) mixing D1 and D2, concentrating under reduced pressure, and lyophilizing.

Preferably, the red sage and safflower extract is prepared by the following method: mixing Saviae Miltiorrhizae radix and Carthami flos, soaking in warm water, and concentrating the filtrate to obtain Saviae Miltiorrhizae radix-Carthami flos concentrated water extract.

Preferably, the red sage and safflower extract is prepared by the following method: mixing the red sage root and the safflower according to the mass ratio of 3:1, soaking twice with 10 times of water at 65 ℃ for 1 hour each time, filtering, combining the filtrates, and concentrating to 1 time of volume to obtain the red sage root-safflower concentrated water extract.

Preferably, the extract of red sage and safflower mainly comprises glucose, galactose, arabinose, fructose, rhamnose, rutinose, rutulose, melibiose, sucrose, trehalose, succinic acid, malonic acid, formic acid, isoleucine, leucine, valine, threonine, alanine, proline, pyroglutamic acid, trigonelline and uridine.

Preferably, the content (mean ± standard deviation) of the red sage and safflower extract is as follows:

the preparation method of the red sage root and safflower extract comprises the following specific steps:

(1) passing the Saviae Miltiorrhizae radix-Carthami flos concentrated water extract through octadecyl silane bonded silica gel open type chromatographic column, respectively eluting with three gradient eluents of 0.1% formic acid water, 0.1% formic acid-water methanol (90:10), and 0.1% formic acid water-methanol (5:95), and washing each gradient with 27, 14 and 1 column volume respectively;

(2) collecting the first 33 column volume fractions as D1 and concentrating and drying under reduced pressure;

(3) collecting 34 th and 35 th column volume fractions, further separating with 0.1% formic acid water (A phase) -methanol (B phase) elution system at 280nm ultraviolet detection wavelength by SB-C18 high performance liquid chromatography column with sample amount of 400 μ L, washing with 95% A phase and 5% B phase at 0-20min flow rate of 10mL/min, and using the same ratio of mobile phase at 21-22min flow rate of 8 mL/min; collecting the flow part obtained in 0-22min as D2;

(4) mixing D1 and D2, concentrating under reduced pressure, and lyophilizing.

The application of the red sage root and safflower extract in preparing the medicine for preventing and relieving/alleviating myocardial ischemia-reperfusion injury.

The myocardial ischemia reperfusion injury refers to myocardial ischemia-induced reperfusion injury caused by open heart surgery, coronary artery bypass surgery, coronary artery transluminal angioplasty, thrombolysis surgery, sudden increase in myocardial medial branch circulation blood volume, and the like.

The invention has the beneficial effects that:

the red sage root and safflower extract effectively screens and retains core functional components through further extraction, reduces the side effect of the medicament, and improves the energy metabolism disorder of ischemia reperfusion heart through inhibiting glucose metabolism and stimulating fatty acid metabolism function; the occurrence of thrombus is prevented by inhibiting the degranulation and aggregation of the platelets to improve hemodynamic parameters; the level of taurine which is an endogenous metabolic marker of oxidative stress resistance is adjusted up, lipid peroxidation is inhibited, and myocardial oxidative stress damage is protected; can also inhibit myocardial inflammation caused by ischemia reperfusion injury through complement system; has obvious effect of relieving myocardial ischemia reperfusion injury of patients with coronary heart disease.

Drawings

FIG. 1 shows the structural formulas of 22 chemical components in the primary component of Chinese medicine;

FIG. 2 is a box line graph of the primary composition of Chinese medicine modulating the metabolite markers in the myocardium;

FIG. 3 is a bar graph of primary component regulatory myocardial lipid markers in a traditional Chinese medicine;

FIG. 4 is a boxplot of metabolite markers in serum regulated by primary components of traditional Chinese medicine;

FIG. 5 bar graph of primary components of Chinese herbs modulating lipid markers in serum.

Detailed Description

To further illustrate the present invention, reference is made to the following examples: