CN110731979A - Application of rhus chinensis total phenolic acid pellets in preparation of medicines for preventing and/or treating cardiovascular diseases - Google Patents
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Abstract
The invention provides application of rhus chinensis total phenolic acid pellets in preparation of a medicine for preventing and/or treating cardiovascular diseases. Experiments prove that the prepared rhus chinensis total phenolic acid pellets can inhibit the elevation of an ST segment of an electrocardiogram and improve the change of T waves, can obviously reduce the myocardial ischemia positive rate and the arrhythmia incidence rate, and show that the rhus chinensis total phenolic acid pellets have obvious prevention and protection effects on cardiovascular diseases, particularly arrhythmia. Therefore, the rhus chinensis total phenolic acid pellets have good application prospect in preparing medicaments for preventing and/or treating cardiovascular diseases, particularly arrhythmia.
Description
Technical Field
The invention belongs to the field of medicine preparation, and particularly relates to application of rhus chinensis total phenolic acid pellets in preparation of medicines for preventing and/or treating cardiovascular diseases, particularly arrhythmia.
Background
Coronary Heart Disease (CHD) is kinds of common cardiovascular diseases, which are short for Coronary atherosclerotic heart disease, important marks of the CHD are atherosclerotic plaques, which can cause chronic stenosis, spasm or obstruction of Coronary arteries with the passage of time, thereby causing myocardial ischemia, hypoxia or infarction to cause heart disease, acute myocardial ischemia is kinds of myocardial ischemia, and means relative or absolute insufficient blood supply of myocardium and myocardial cell hypoxia caused by various reasons.
Arrhythmia is the most common complex and dangerous cardiovascular disease or complication, is causing high mortality of cardiovascular disease, can be singly attacked, and can also be accompanied with other cardiovascular diseases, the clinical manifestations of arrhythmia are palpitation, shortness of breath, chest distress, hypodynamia, insomnia, dreaminess and other symptoms, and the existing medicines for treating arrhythmia include amiodarone, lidocaine, Diaoxinxuekang capsules and the like.
However, most of the existing medicines for treating cardiovascular diseases such as coronary heart disease and arrhythmia have side effects, and the traditional Chinese medicines are concerned with due to relatively small side effects, in recent years, the research on the cardiovascular diseases by the traditional Chinese medicines is increasing, people have better effects of preventing and treating the cardiovascular diseases by applying the traditional Chinese medicines, and corresponding traditional Chinese medicine preparations are continuously developed.
However, the basis of the drug effect substance of rhus chinensis for radically treating coronary heart disease is not clear up till now. So far, scholars at home and abroad also have insufficient research on chemical components of rhus chinensis roots, so that the research on the chemical components in the rhus chinensis roots is of great significance in finding medicines capable of effectively treating cardiovascular diseases (such as coronary heart disease, acute myocardial ischemia, arrhythmia, heart disease caused by anoxia or infarction, and the like).
Disclosure of Invention
The invention aims to provide application of rhus chinensis total phenolic acid pellets in preparation of medicines for preventing and/or treating cardiovascular diseases, in particular arrhythmia.
The invention provides application of types of rhus chinensis total phenolic acid pellets in preparation of medicines for preventing and/or treating cardiovascular diseases.
, the cardiovascular diseases include coronary heart disease, myocardial ischemia, and heart disease.
Further , the myocardial ischemia is acute myocardial ischemia.
Further , the cardiovascular disease is arrhythmia.
, the medicine can reduce ST segment change of electrocardiogram and reduce arrhythmia incidence.
the preparation method comprises collecting total phenolic acid of Rhus chinensis Mill, and making into pellet.
, the preparation method of the rhus chinensis total phenolic acid pellet comprises the following steps:
(1) adding organic solvent into Rhus chinensis Mill, extracting, and filtering to obtain filtrate;
(2) eluting and purifying the filtrate obtained in the step (1) by using macroporous resin as an adsorption filler: collecting the eluted part of the alcohol solvent by using water as an eluent and using the alcohol solvent as an eluent, concentrating, freezing and drying to obtain the rhus chinensis total phenolic acid;
(3) mixing the rhus chinensis total phenolic acid obtained in the step (2), a balling promoter, a pore-forming agent, an anti-sticking agent and a wetting agent, and preparing the rhus chinensis total phenolic acid pellets in a low-temperature extrusion spheronizer.
, in the step (1), the rhus chinensis is rhus chinensis tubers, the organic solvent is ethanol, preferably 70% ethanol, and the mass-volume ratio of the rhus chinensis to the organic solvent is 1 (10-30) g/mL;
and/or the extraction temperature is 70-90 ℃, and the extraction time is 60-120 minutes; preferably, the mass volume ratio of the rhus chinensis to the ethanol is 1: 20 g/mL; the extraction temperature is 80 ℃, and the extraction time is 90 minutes.
, in the step (2), the using amount of the water is 4BV, the using amount of the alcohol solvent is 4BV, and the alcohol solvent is 50% ethanol;
and/or the macroporous resin is SP825L macroporous resin, the sample loading amount is 0.35g rhus chinensis/g macroporous resin, and the flow rate of the eluent is 2 BV.h-1。
, in step (3), the balling accelerant is a cellulose balling accelerant, the pore-forming agent is a water-soluble pore-forming agent, the anti-sticking agent is an inorganic non-metal oxide, and the wetting agent is an alcohol substance, preferably, the balling accelerant is microcrystalline cellulose, the pore-forming agent is lactose, the anti-sticking agent is silicon dioxide, and the wetting agent is 60% ethanol;
the process conditions of the low-temperature extrusion rounding machine are as follows: extrusion speed 43r min-1Speed of rounding is 1800 r.min-1And the rounding time is 4 min.
, in the step (3), the mass volume ratio of the rhus chinensis total phenolic acid to the balling promoter to the pore-forming agent to the anti-sticking agent to the wetting agent is 45g to 94.5g to 10.5g to 6.75g to 110 mL.
In the present invention, the pellets are spherical or spheroidal solid dosage forms having a diameter of less than 2.5mm, and can be filled into capsules, compressed into tablets, or made into other formulations.
The experimental result shows that the rhus chinensis total phenolic acid pellet prepared by the invention can inhibit the elevation of an ST segment of an electrocardiogram and improve the change of T wave, can obviously reduce the myocardial ischemia positive rate and the arrhythmia incidence rate, and particularly has the effect of reducing the arrhythmia incidence rate of myocardial ischemia rats which is obviously superior to that of a YFM total phenolic acid group. The rhus chinensis total phenolic acid pellets have obvious prevention and protection effects on cardiovascular diseases, particularly arrhythmia. Therefore, the rhus chinensis total phenolic acid pellets have good application prospect in preparing medicaments for preventing and/or treating cardiovascular diseases, particularly arrhythmia.
Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.
The present invention will be described in further detail by way of examples , but it should not be construed that the scope of the above subject matter is limited to the examples described below.
Drawings
Fig. 1 shows typical electrocardiograms of each group.
Detailed Description
The raw materials and equipment used in the invention are known products and are obtained by purchasing commercial products.
Example 1 preparation of crude extract of Rhus chinensis Mill
Taking Rhus chinensis Mill tuber (5g), adding 70% ethanol (100mL), extracting at 80 deg.C for 90min twice, filtering, collecting filtrate, concentrating, and drying to obtain Rhus chinensis Mill crude extract.
Example 2 preparation of Total phenolic acids of Rhus chinensis Mill
Collecting Rhus chinensis Mill tuber (5g), adding 70% ethanol (100mL), extracting at 80 deg.C for 90min twice, and filtering to obtain filtrate. Using SP825L macroporous resin (diameter-height ratio of 1:5) as adsorption filler, and adding 1 BV.h into the filtrate-1The sample was applied at a flow rate (amount of sample: 0.35g crude drug/g dry resin). Then 4BV of water is firstly usedEluent at 2 BV.h-1Removing impurities, eluting with 4BV 50% ethanol at 2 BV.h-1Eluting, collecting 50% ethanol eluate, concentrating, and freeze drying to obtain total phenolic acid of Rhus chinensis Mill.
Example 3 preparation of Total phenolic acid pellets of Rhus chinensis Mill
The prescription composition is as follows: 45g of rhus chinensis total phenolic acid, 94.5g of microcrystalline cellulose, 10.5g of lactose, 6.75g of silicon dioxide and 60% ethanol (110 +/-2) mL.
Weighing the raw materials according to the formula, and preparing the rhus chinensis total phenolic acid pellet by adopting a Mini-250 experimental type low-temperature extrusion spheronizer, wherein the preparation process comprises the following steps: extrusion speed 43r min-1Speed of rounding is 1800 r.min-1And the rounding time is 4 min.
The beneficial effects of the present invention are demonstrated by the following experimental examples.
Experimental example 1 evaluation of therapeutic Effect of Rhus chinensis Total phenolic acid on myocardial ischemia rats
1 Instrument and reagent
1.1 instruments
BL-420A biological function test system (Chengdotai alliance software Co., Ltd.), XS105 electronic analytical balance (Mettler-Torledo instruments Co., Ltd.), a desk-top refrigerated centrifuge (Beckmann Kulter, USA), 5418-Eppendorf centrifuge (Elegde GmbH, Germany), HH-4 digital display thermostat water bath (Waals, U.S. China, Ltd.), a vortex mixer shaker (Hai , Linebel instruments Co., Ltd.), a multifunctional microplate reader (Infinite, Austria), UV-1800 ultraviolet spectrophotometer (Shimadzu).
1.2 reagent
Diaoxinxuekang capsules (specification: 100mg, Duoduo pharmaceutical group Co., Ltd., lot # 1801051), nifedipine (specification: 10mg, Huazhou Chinese medicinal industry Co., Ltd., lot # 20180304), crude extract of Rhus chinensis (prepared in example 1), total phenolic acids of Rhus chinensis (prepared in example 2), pellets of total phenolic acids of Rhus chinensis (prepared in example 3, drug loading amount: 28.7%), pituitrin injection (specification: 6U: 2mL, Shanghai medicine Biochemical industry Co., Ltd., lot # 1704201), 0.9% sodium chloride injection (specification: 500 mL: 4.5g, King Fu pharmaceutical Co., Ltd., Fuzhou, lot # 161215A43), Lactate Dehydrogenase (LDH) kit (96T), and Creatine Kinase (CK) kit (50T) (Ministry of biological engineering research).
1.3 animals
SPF grade SD rats, male, with a body mass of 250 ± 20g, purchased from shanghai slyke laboratory animals llc, laboratory animal license number: SCXK (Shanghai) 2012-0002.
2 method of experiment
2.1 animal grouping and modeling
SD rats are fed for 5 days adaptively, 10% chloral hydrate is used for anesthesia, the electrocardiogram of the rats is measured, 56 rats with normal limb II-lead electrocardiogram (the S point is not higher than the baseline after anesthesia) are selected, and the selected rats are randomly divided into 7 groups: normal group, model group, positive medicine Di' ao Xinxuekang capsule (12.26 mg. mL)-1) Nifedipine (1.23 mg. mL) as positive drug-1) Crude extract of rhus chinensis (named YFM crude extract group, 1.84 g.mL)-1) Rhus chinensis total phenolic acid (named as YFM total phenolic acid group, 39 mg/mL)-1) Rhus chinensis total phenolic acid pellets (named YFM pellet group, 136 mg/mL)-1) Each group had 8. From the 6 th day, the normal group and the model group are filled with stomach and water for 10 days, and the rest are continuously filled with stomach and water for 10 days; the volume of the drug (or water) is 5 mL/kg-1After 2h of the last administrations, 1 U.kg of posterior pituitrin (pit) was administered by sublingual intravenous injection-1The method comprises establishing an acute myocardial ischemia model, and recording a II-lead electrocardiogram by using a BL-420A biological function experiment system. When sublingual intravenous injection pit causes myocardial ischemia of rats, the phase 2 can be generally divided by observing the change of ST segment of electrocardiogram, 5-30 s T waves are increased after the phase 1 injection, and ST segment is raised (> 0.1 mV); 30s to minutes after injection at stage 2, ST shift downward, T wave decrease, plateau, bilateral or inverted, heart rate slow, P-R and Q-T interval prolonged, ischemic change at stage 1 or stage 2 becomes positive, otherwise negative. The ST segment is raised to be more than 0.1mV 30s after the experimental model group injects the posterior pituitrin, the ST segment is moved downwards after 30s, and the T wave is low and flat or inverted, which indicates that the molding is successful.
2.2 index detection
2.2.1 measurement of changes in electrocardiography in rats with myocardial ischemia
10% chloral hydrate is used for intraperitoneal injection anesthesia (0.3mL 100 g)-1) Fixing the anesthesia apparatus on an operation table board after anesthesia, inserting a needle for injection under the skin of four limbs, connecting the needle with limb leads by using an alligator clip, and recording a II-lead electrocardiogram of the rat limb; taking PP connecting line as a baseline, recording electrocardiogram ST segment values of 0, 5, 10, 15, 30s and 1, 5 and 10min before and after injection of the pit, and analyzing and comparing ST segment changes before and after injection of the pit of each group.
patients with wide or abnormal QRS waves, no inverse P wave or inverted P wave, arrhythmia and the like appear in the electrocardiogram, and the patients with arrhythmia and the like are recorded as arrhythmia, and the patients with patients with arrhythmia are recorded as positive myocardial ischemia, wherein J point is increased by more than 1.5mV, T wave is decreased by more than 50%, the patients are subjected to bidirectional inversion, ST segment is increased by 0.1mV or pressed by 0.5mV, and the fluctuation condition of ST segment and the change of heart rate before and after sublingual intravenous injection of pituitrin are observed and recorded.
2.2.2 detection of Biochemical indicators in serum of rats with myocardial ischemia
Lactate Dehydrogenase (LDH) and phosphocreatine kinase (CK) in serum are important indexes for evaluating myocardial damage, the level of LDH and CK can measure the degree of myocardial damage, myocardial cell membranes are damaged by acute myocardial ischemia and hypoxia, and then a large amount of oxygen radicals are generated, so that LDH and CK which are markers of myocardial damage are released into blood in a large amount, and the levels of LDH and CK in serum are obviously increased.
After measuring electrocardiogram, 5mL of blood is taken from abdominal aorta of rat, left for 2h at room temperature, centrifuged for 10min (3500 r.min)-1) Carefully remove the serum into an EP tube and store it at-80 ℃ for further use. During measurement, serum is taken out, is quickly thawed, and is strictly operated according to the kit steps to measure the activity of LDH and CK in the serum.
2.3 statistical treatment
Statistical analysis of the data was performed using SPSS21 software to analyze the dataThe comparison among the groups adopts one-factor variance analysis, data are compared pairwise by an LSD method, and when P is less than 0.05, the significance difference is judged.
3 results of the experiment
3.1 myocardial ischemia rat electrocardiogram ST segment variation
The ST segment of the first 30s of the rats in the model group after being injected with the pituitrin is obviously raised, the ST segment value is raised to be more than 0.1mV, and the rats have significant difference (P is less than 0.01) compared with the normal group, which indicates that the modeling is successful, and the typical electrocardiograms of the rats in each group are obtained by real-time monitoring and observation after the modeling, see figure 1. through statistical analysis, as can be seen from table 1, the ST segment of each group before the sublingual intravenous injection of pit is carried out for the modeling has no significant difference, and indicates that the administration by gastric lavage has no influence on the electrocardiograms of the normal rats, the observation of the electrocardiograms of 5, 10 and 15s after the injection of pit shows that although the ST segment of each administration group is raised, the raising amplitude of the ST segment at the stage is reduced compared with the model group, the ST segment raising value is less than 0.1mV, and compared with the model group, except for YFM crude extract, each group has significant difference.
From table 1, it can be found that the maximum change amount of the ST segment of the YFM crude extract is 0.080mV, the maximum change amount of the ST segment of the YFM total phenolic acid group is 0.062mV, and the maximum change amount of the ST segment of the YFM pellet group is 0.059 mV., the rhus chinensis crude extract has definite effect on relieving the pit-induced acute myocardial ischemia, and the rhus chinensis total phenolic acid pellets have obvious protective effect on rats with acute myocardial ischemia, wherein the protective effect of the rhus chinensis total phenolic acid pellets is most obvious, and the ST segment change of the rats with acute myocardial ischemia can be quickly reduced.
Note a: p < 0.05, P < 0.01 compared to model group.
3.2 Heart Rate Change in rats with myocardial ischemia
The results are shown in Table 2. After sublingual intravenous injection of pit, the heart rate of rats in each group (except a normal group) is gradually reduced along with the time extension within 10min, the heart rate reduction of the model group at each time point is particularly obvious, and the heart rate reduction is remarkably different from that of the normal group (P < 0.01); although the heart rate of each administration group is slowed down, the heart rate of each administration group is not significantly different from that of a normal group (P is more than 0.05). When pit 5s is injected, the heart rate reduction amplitude of the model group is large, and the heart rate reduction amplitude is remarkably different from that of the normal group (P is less than 0.01); at 10s, the heart rate of each administration group was reduced except for the YFM crude extract, but the reduction was smaller than that of the model group, and the heart rate was significantly higher than that of the model group (P < 0.05 or 0.01). The administration groups can reduce the arrhythmia incidence rate of the myocardial ischemia rats, wherein the effect of the YFM pellet group on reducing the arrhythmia incidence rate of the myocardial ischemia rats is obviously superior to that of the YFM total phenolic acid group (P is less than 0.05), and the effect is most similar to that of the positive drug group.
Note: p < 0.05, P < 0.01, compared to model group;
comparing YFM micropill with YFM total phenolic acid, # P is less than 0.05.
3.3 Biochemical index results of serum from rats with myocardial ischemia
The results are shown in Table 3. Compared with the normal group, the activity level of LDH and CK in the serum of the rat of the sublingual intravenous injection pit model group is obviously increased, and the significant difference is realized (P is less than 0.01). Compared with the model group, the activity levels of LDH and CK in the serum of each administration group are reduced, and the significant difference is achieved (P is less than 0.01). And the reduction effect of the YFM total phenolic acid and the YFM crude extract on the activity levels of LDH and CK is better than that of the YFM pellet, so that the regulation effect of the YFM total phenolic acid and the YFM crude extract on the biochemical indexes of LDH and CK in the serum of a myocardial ischemia rat is better than that of the YFM pellet.
TABLE 3 notesMeasurement result of serum biochemical index of rat with acute myocardial ischemia induced by Rapid pit: (n=8)
Group of | Dosage per g.kg-1 | LDH/U·mL-1 | CK/U·L-1 |
Normal group | — | 6194.42±1347.27** | 2.61±0.32** |
Model set | — | 9871.87±2324.99 | 9.08±1.44 |
Di' ao Xinxuekang capsule | 0.0613 | 7003.24±1220.02** | 3.62±1.07** |
Nifedipine | 0.00613 | 5075.10±2551.07** | 3.30±1.09** |
Crude YFM extract | 9.2 | 5712.58±1706.16** | 3.40±0.76** |
YFM Total phenolic acid | 0.195 | 4631.41±1063.14** | 4.30±1.03** |
YFM pellet | 0.68 | 5866.71±1471.00** | 4.45±1.16** |
Note a: p < 0.05 compared to the model group,
b, note: p < 0.01 compared to model group.
In conclusion, the rhus chinensis total phenolic acid pellet prepared by the invention can inhibit the elevation of the ST segment of an electrocardiogram and improve the change of T wave, can obviously reduce the myocardial ischemia positive rate and the arrhythmia incidence rate, and particularly has the effect of reducing the arrhythmia incidence rate of myocardial ischemia rats which is obviously better than that of the YFM total phenolic acid group. The rhus chinensis total phenolic acid pellets have obvious prevention and protection effects on cardiovascular diseases, particularly arrhythmia. Therefore, the rhus chinensis total phenolic acid pellets have good application prospect in preparing medicaments for preventing and/or treating cardiovascular diseases, particularly arrhythmia.
Claims (10)
1. Application of total phenolic acid pellet of Rhus chinensis Mill in preparing medicine for preventing and/or treating cardiovascular diseases is provided.
2. Use according to claim 1, characterized in that: the cardiovascular diseases include coronary heart disease, myocardial ischemia, and heart disease.
3. Use according to claim 2, characterized in that: the myocardial ischemia is acute myocardial ischemia.
4. Use according to claim 1, characterized in that: the cardiovascular disease is arrhythmia.
5. Use according to claim 1, characterized in that: the medicine can reduce the change of ST segment of electrocardiogram and reduce the incidence rate of arrhythmia.
6. The use of any one of of claims 1-5, wherein the total phenolic acid of Rhus chinensis Mill is prepared by collecting total phenolic acid of Rhus chinensis Mill, and making into pellet.
7. Use according to claim 6, characterized in that: the preparation method of the rhus chinensis total phenolic acid pellet comprises the following steps:
(1) adding organic solvent into Rhus chinensis Mill, extracting, and filtering to obtain filtrate;
(2) eluting and purifying the filtrate obtained in the step (1) by using macroporous resin as an adsorption filler: collecting the eluted part of the alcohol solvent by using water as an eluent and using the alcohol solvent as an eluent, concentrating, freezing and drying to obtain the rhus chinensis total phenolic acid;
(3) mixing the rhus chinensis total phenolic acid obtained in the step (2), a balling promoter, a pore-forming agent, an anti-sticking agent and a wetting agent, and preparing the rhus chinensis total phenolic acid pellets in a low-temperature extrusion spheronizer.
8. Use according to claim 7, characterized in that: in the step (1), the rhus chinensis is rhus chinensis tubers; the organic solvent is ethanol, preferably 70% ethanol; the mass volume ratio of the rhus chinensis to the organic solvent is 1: (10-30) g/mL;
and/or the extraction temperature is 70-90 ℃, and the extraction time is 60-120 minutes; preferably, the mass volume ratio of the rhus chinensis to the ethanol is 1: 20 g/mL; the extraction temperature is 80 ℃, and the extraction time is 90 minutes;
and/or in the step (2), the using amount of the water is 4BV, the using amount of the alcohol solvent is 4BV, and the alcohol solvent is 50% ethanol;
and/or the macroporous resin is SP825L macroporous resin, the sample loading amount is 0.35g rhus chinensis/g macroporous resin, and the flow rate of the eluent is 2 BV.h-1。
9. Use according to claim 8, characterized in that: in the step (3), the balling accelerant is a cellulose balling accelerant, the pore-forming agent is a water-soluble pore-forming agent, the anti-sticking agent is an inorganic non-metal oxide, and the wetting agent is an alcohol substance, preferably, the balling accelerant is microcrystalline cellulose, the pore-forming agent is lactose, the anti-sticking agent is silicon dioxide, and the wetting agent is 60% ethanol;
the process conditions of the low-temperature extrusion rounding machine are as follows: extrusion speed 43r min-1Speed of rounding is 1800 r.min-1And the rounding time is 4 min.
10. Use according to claim 9, characterized in that: in the step (3), the mass volume ratio of the rhus chinensis total phenolic acid to the balling promoter to the pore-forming agent to the anti-sticking agent to the wetting agent is 45 g: 94.5 g: 10.5 g: 6.75 g: 110 mL.
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Cited By (1)
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CN111297926A (en) * | 2019-03-22 | 2020-06-19 | 福建中医药大学 | Rhus chinensis total phenolic acid pellet and preparation method thereof |
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2019
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CN111297926A (en) * | 2019-03-22 | 2020-06-19 | 福建中医药大学 | Rhus chinensis total phenolic acid pellet and preparation method thereof |
CN111297926B (en) * | 2019-03-22 | 2022-04-05 | 福建中医药大学 | Rhus chinensis total phenolic acid pellet and preparation method thereof |
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