CN109091532B - Application of Tongguan capsule in preparation of medicine for treating hypertension and heart failure - Google Patents

Abstract

The invention relates to the field of medicines, in particular to a new application of a Tongguan capsule, and in particular relates to an application of the Tongguan capsule in preparation of a hypertensive heart failure medicine, wherein the main active ingredients of the Tongguan capsule comprise the following raw material medicines in percentage by weight: 40-60% of astragalus membranaceus, 20-50% of salvia miltiorrhiza and 5-25% of leech. Experiments prove that the coronary dredging capsule has good treatment and relief effects on improving ventricular hypertrophy, increasing cardiac contractility, delaying ventricular remodeling and treating and improving hypertensive heart failure, and the treatment effect of the coronary dredging capsule is superior to that of metoprolol; moreover, the capsule has less medicinal flavor, reliable curative effect determination, definite medicinal effect components and less adverse reaction, and can obviously improve the prognosis and the life quality of patients.

Description

Application of Tongguan capsule in preparation of medicine for treating hypertension and heart failure

Technical Field

The invention relates to the field of traditional Chinese medicines, in particular to application of a Tongguan capsule in preparation of a medicine for treating hypertension and heart failure.

Background

Hypertension, if poorly controlled for long periods of time, can cause structural and functional changes in the heart, known as hypertensive heart disease, including: early-stage hypodiastolic function of the left ventricle and left ventricular hypertrophy, the progressive occurrence of hypomyocardial contractility finally leads to heart failure, and 70 percent of the heart failure is caused by hypertension. Currently, the heart failure mechanism caused by hypertension is not completely clear, the basic mechanism leading to the development of heart failure is myocardial remodeling, and hypertension can cause myocardial hypertrophy, myocardial fibrosis and myocardial reduction with contractility (gull and the like, the prevention and treatment of hypertensive heart failure [ J ]. Chinese medical journal, 2012,47(7): 6-9.). Under the action of certain inducement (such as infection, emotional agitation, mental stress, drinking and the like), patients with hypertension are easy to have acute heart failure, and if the patients are not actively treated, the death rate is very high. Moreover, hypertension heart failure is more common than heart failure caused by endocardial infection or arrhythmia, has serious symptoms and is difficult to reverse, and most of the hypertension heart failure cannot be treated by using antihypertensive drugs or cardiotonic drugs alone.

The development of hypertensive heart failure is a progressive process that once initiated, can progress itself even in the stable phase of the clinic without new myocardial damage. In recent years, the medicine and surgical treatment of heart failure are greatly improved, a more systematic treatment scheme is provided clinically, the morbidity and the mortality cannot be reduced ideally, and the large-scale clinical application of the medicine and the surgical treatment in the heart failure treatment is limited due to the side effect of western medicines and relatively high treatment cost, so that the long-term clinical treatment is difficult to complete. Therefore, the seeking of a treatment method and a medicine with definite clinical curative effect and small safety and toxic and side effects are always paid attention to by clinicians in various countries, so that people turn to the traditional medical treatment method of traditional Chinese medicine.

The patent CN101647856A discloses a medicine for treating coronary heart disease, which is particularly suitable for treating patients after coronary heart disease intervention, the active ingredients of the medicine comprise astragalus root, salvia miltiorrhiza and leech, the effect of the medicine for treating stable angina pectoris of coronary heart disease is obvious, and the most common reason of stable angina pectoris of coronary heart disease is that myocardial oxygen consumption is increased on the basis of coronary artery fixed stenosis, and the pathological basis is coronary artery atherosclerotic stenosis. The medicine for treating coronary heart disease provided by the patent can effectively treat and integrally regulate patients after coronary heart disease intervention by acting on a plurality of targets of atherosclerosis and restenosis such as vascular smooth muscle cells, endothelial cells, platelets, a blood coagulation-fibrinolysis system, blood fat, inflammatory factors and the like, but the coronary-through capsule is not reported to be used for treating hypertension and heart failure at present.

Disclosure of Invention

The invention aims to provide a new application of a Tongguan capsule, namely the capsule for preparing a medicine for treating hypertension and heart failure.

According to one aspect of the invention, the invention provides a new application of the Tongguan capsule, namely the application of the Tongguan capsule in preparing a medicine for treating hypertension and heart failure, wherein the main active ingredients of the Tongguan capsule comprise the following raw material medicines in percentage by weight: 40-60% of astragalus membranaceus, 20-50% of salvia miltiorrhiza and 5-25% of leech.

In some embodiments, the main active ingredients in the Tongguan capsule are preferably 45-55% of astragalus, 25-35% of salvia miltiorrhiza and 10-20% of leech; the optimal mixture ratio is as follows: 50% of astragalus root, 35% of salvia miltiorrhiza and 15% of leech.

In some embodiments, the medicament consists of the main active ingredient in the Tongguan capsule and pharmaceutically acceptable excipients, and can be prepared into capsules, soft capsules, granules, tablets, dripping pills or oral liquid for patients to take.

The medicine for treating hypertensive heart failure can adopt the corresponding existing preparation method according to the specification requirements of specific preparations, wherein the recommended method is as follows:

the method comprises the following steps:

the preparation method of the medicine for treating hypertensive heart failure comprises the following steps:

(1) weighing Saviae Miltiorrhizae radix and Hirudo according to a certain proportion, cleaning, oven drying, pulverizing, and sieving with 120 mesh sieve to obtain Saviae Miltiorrhizae radix-Hirudo powder;

(2) weighing radix astragali according to a ratio, adding 10 times of water each time, decocting for three times, boiling for 1h, 45min and 45min each time, filtering through 60-mesh filter cloth while hot, combining the three filtrates, concentrating until the water content is less than or equal to 8%, cooling to room temperature, slowly adding 60% ethanol by volume, refrigerating and standing for 24 h-48 h, filtering, concentrating until the relative density is 1.10-1.20 at 20 ℃, and recovering ethanol to obtain radix astragali extract;

(3) and (3) fully and uniformly mixing the salvia-leech powder prepared in the step (1) and the astragalus extract prepared in the step (2), drying, crushing, sieving by a 120-mesh sieve, adding a pharmaceutically acceptable excipient, and preparing into capsules, granules, tablets or soft capsules.

The method 2 comprises the following steps:

the preparation method of the medicine for treating hypertensive heart failure comprises the following steps:

(1) weighing the astragalus and the salvia miltiorrhiza according to the proportion, adding 10 times of water each time, decocting for three times, boiling for 1h, 45min and 45min each time respectively, filtering through 60-mesh filter cloth while the mixture is hot, combining the three filtrates, concentrating until the water content is less than or equal to 8%, cooling to room temperature, slowly adding ethanol with the volume ratio of 60%, refrigerating and standing for 24 h-48 h, filtering, concentrating until the relative density is 1.10-1.20 at 20 ℃, and recovering the ethanol to obtain astragalus-salvia miltiorrhiza extract;

(2) weighing leeches according to the proportion, cleaning, drying, crushing, and sieving with a 120-mesh sieve to obtain leech powder;

(3) and (3) fully and uniformly mixing the extract prepared in the step (1) and the leech powder prepared in the step (2), drying, crushing, sieving by a 120-mesh sieve, and adding a pharmaceutically acceptable excipient to prepare capsules, granules, tablets or soft capsules.

The method 3 comprises the following steps:

the preparation method of the medicine for treating hypertensive heart failure comprises the following steps:

(1) weighing the salvia miltiorrhiza according to the proportion, adding 7 times of ethanol, heating and refluxing for 1.5h, filtering, adding 50% filtrate of ethanol into dregs, heating and refluxing for 1h, filtering, combining the filtrates, concentrating to the relative density of 1.10-1.20 at 20 ℃, and recovering ethanol to obtain salvia miltiorrhiza extract;

(2) weighing astragalus membranaceus according to a ratio, adding the astragalus membranaceus into the salvia miltiorrhiza dregs, adding 10 times of water each time, decocting for three times, boiling for 1 hour, 45 minutes and 45 minutes respectively each time, filtering through 60-mesh filter cloth while the decoction is hot, combining filtrate obtained in the three times, concentrating the filtrate under reduced pressure until the water content is less than or equal to 8%, cooling to room temperature, slowly adding 60% ethanol in volume ratio, refrigerating and standing for 24 hours to 48 hours, filtering, concentrating until the relative density is 1.10-1.20 at 20 ℃, and recovering the ethanol to obtain an astragalus membranaceus extract;

(3) weighing leeches according to the proportion, cleaning, drying, crushing, and sieving with a 120-mesh sieve to obtain leech powder;

(4) and (3) fully and uniformly mixing the extract prepared in the steps (1) and (2) and the leech powder prepared in the step (3), drying, crushing, sieving by a 120-mesh sieve, and adding a pharmaceutically acceptable excipient to prepare capsules, granules, tablets or soft capsules.

The method 4 comprises the following steps:

the preparation method of the medicine for treating hypertensive heart failure comprises the following steps:

(1) weighing the astragalus and the salvia according to the proportion, adding 10 times of water each time, decocting for three times, boiling for 1h, 45min and 45min each time respectively, filtering through 60-mesh filter cloth while the mixture is hot, combining the three filtrates, concentrating until the water content is less than or equal to 8%, cooling to room temperature, slowly adding ethanol with the volume ratio of 60%, refrigerating and standing for 24 h-48 h, filtering, concentrating, and recovering the ethanol to obtain astragalus-salvia concentrated solution;

(2) weighing Hirudo according to a certain proportion, adding 10 times of water, decocting for three times, boiling for 1 hr, 45min and 45min each time, filtering with 60 mesh filter cloth while hot, mixing filtrates, and concentrating the filtrate under reduced pressure to obtain Hirudo concentrated solution;

(3) and (3) uniformly mixing the concentrated solutions prepared in the step (1) and the step (2), adding pharmaceutically acceptable excipient, and preparing the oral liquid or the dropping pill.

Animal experiments and clinical use results prove that the coronary dredging capsule has good treatment and relief effects on improving ventricular hypertrophy, increasing cardiac contractility, delaying ventricular remodeling and treating and improving hypertensive heart failure, and the treatment effect of the coronary dredging capsule is superior to that of metoprolol; moreover, the capsule has less medicinal flavor, reliable curative effect determination, definite medicinal effect components and less adverse reaction, and can obviously improve the prognosis and the life quality of patients.

Drawings

FIG. 1 is a graph of the survival of rats when animals were modeled;

FIG. 2 is an echocardiogram of four groups of rats;

FIG. 3 is a left ventricular Ejection Fraction (EF) measurement of four groups of rats, where # indicates P < 0.05 compared to the control group and # indicates P < 0.05 compared to the ISO group;

FIG. 4 is a left ventricular short axis shortening (FS) measurement of four groups of rats, where # indicates P < 0.05 compared to the control group and # indicates P < 0.05 compared to the ISO group;

FIG. 5 shows the lung-diameter ratio measurements of four groups of rats, wherein P < 0.01 in comparison with the control group and # indicates P < 0.01 in comparison with the ISO group;

fig. 6 shows the results of measurements of the ratios of the heart to shin of four groups of rats, where P < 0.001 compared to the control group and P < 0.05 compared to the ISO group;

FIG. 7 is a graph of lung volume ratio measurements for four groups of rats, where P < 0.01 compared to the control group;

FIG. 8 is a graph of cardiac body ratio measurements for four groups of rats, where P < 0.001 compared to the control group and # indicates P < 0.01 compared to the ISO group;

FIG. 9 is a graph showing the gross appearance of four groups of rat hearts and the results of HE staining of cardiomyocytes;

FIG. 10 is a graph showing results of sirius red staining of four groups of rat cardiac muscle cells;

FIG. 11 is a graph comparing the measurement results of the cross-sectional area of cardiomyocytes after one week administration to four groups of rats, where P < 0.001 compared to the control group and # indicates P < 0.001 compared to the ISO group;

FIG. 12 is a graph comparing the results of measurement of fibrosis of heart tissues after administration for one week in four groups of rats, wherein ×) represents P < 0.01 as compared with the control group, # represents P < 0.05 as compared with the ISO group, and # represents P < 0.05 as compared with the ISO group;

FIG. 13 is an electrophoretogram of CaMKII delta and MAPKs pathway proteins of ISO group, MET group and TG group;

FIG. 14 is a graph of the level of ERK phosphorylation, where P < 0.05 compared to control;

FIG. 15 is a graph of the results of the ratio of phosphorylated P38 to unactivated P38, wherein # represents P < 0.05 compared to the control group and # represents P < 0.05 compared to the ISO group;

FIG. 16 is a graph showing the results of the ratio of phosphorylated JNK to non-activated JNK, wherein P < 0.05 compared to the control group and P < 0.05 compared to the ISO group;

FIG. 17 is a graph of CaMKII delta expression, where P < 0.05 compared to the control group and P < 0.05 compared to the ISO group are indicated;

FIG. 18 is an electrophoretogram of ISO-, MET-and TG-group HDAC4, MEF2 and GATA4 pathway proteins;

FIG. 19 is a graph of HDAC4 expression, wherein P < 0.01 in comparison to control, # indicates P < 0.05 in comparison to control, and # indicates P < 0.01 in comparison to ISO;

FIG. 20 is a graph showing the expression of MEF2, wherein P < 0.05 in comparison with the control group and P < 0.05 in comparison with the ISO group.

Detailed Description

The present invention will be described in further detail with reference to the accompanying drawings. Unless otherwise specified, the reagents used in the following tests are commercially available.

Example 1

The preparation method of the Tongguan capsule for preparing the medicine for treating hypertension and heart failure comprises the following steps:

(1) weighing the raw material medicines of astragalus, salvia miltiorrhiza and leech respectively, wherein the weight percentages of the raw material medicines are as follows: astragalus root: red sage root: leech 50%: 35%: 15 percent;

(2) cleaning Saviae Miltiorrhizae radix and Hirudo, oven drying, pulverizing, and sieving with 120 mesh sieve to obtain Saviae Miltiorrhizae radix-Hirudo powder;

(3) adding 10 times of water into astragalus each time, decocting for three times, boiling for 1 hour, 45 minutes and 45 minutes each time, filtering through 60-mesh filter cloth while the astragalus is hot, combining the three filtrates, concentrating until the water content is less than or equal to 8%, cooling to room temperature, slowly adding ethanol with the volume ratio of 60%, refrigerating and standing for 24-48 hours, filtering, concentrating until the relative density is 1.10-1.20 at 20 ℃, and recovering ethanol to obtain astragalus extract;

(4) and (3) fully and uniformly mixing the red sage root-leech powder prepared in the step (2) and the astragalus extract prepared in the step (3), drying, crushing, sieving by a 120-mesh sieve, adding a pharmaceutically acceptable excipient, and preparing into capsules.

Example 2

The preparation method of the Tongguan capsule for preparing the medicine for treating hypertension and heart failure comprises the following steps:

(1) weighing the raw material medicines of astragalus, salvia miltiorrhiza and leech respectively, wherein the weight percentages of the raw material medicines are as follows: astragalus root: red sage root: leech 60%: 20%: 20 percent;

(2) mixing the astragalus and the salvia, adding 10 times of water each time, decocting for three times, boiling for 1h, 45min and 45min each time respectively, filtering through 60-mesh filter cloth while the mixture is hot, combining the three filtrates, concentrating until the water content is less than or equal to 8%, cooling to room temperature, slowly adding 60% ethanol by volume, refrigerating and standing for 24 h-48 h, filtering, concentrating until the relative density is 1.10-1.20 at 20 ℃, and recovering ethanol to obtain astragalus-salvia extract;

(3) cleaning Hirudo, oven drying, pulverizing, and sieving with 120 mesh sieve to obtain Hirudo powder;

(4) and (4) fully and uniformly mixing the extract prepared in the step (3) and the leech powder prepared in the step (4), drying, crushing, sieving by a 120-mesh sieve, and adding a pharmaceutically acceptable excipient to prepare granules.

Example 3

The preparation method of the Tongguan capsule for preparing the medicine for treating hypertension and heart failure comprises the following steps:

(1) weighing the raw material medicines of astragalus, salvia miltiorrhiza and leech respectively, wherein the weight percentages of the raw material medicines are as follows: astragalus root: red sage root: leech is 40%: 50%: 10 percent;

(2) adding 7 times of ethanol into the salvia miltiorrhiza, heating and refluxing for 1.5h, filtering, adding 50% of ethanol into dregs, heating and refluxing for 1h, filtering, combining the filtrates, concentrating to the relative density of 1.10-1.20 at 20 ℃, and recovering the ethanol to obtain salvia miltiorrhiza extract;

(3) adding astragalus into the salvia miltiorrhiza residue, adding 10 times of water each time, decocting for three times, boiling for 1h, 45min and 45min respectively each time, filtering through 60-mesh filter cloth while the decoction is hot, combining the three filtrates, concentrating the filtrate under reduced pressure until the water content is less than or equal to 8%, cooling to room temperature, slowly adding ethanol with the volume ratio of 60%, refrigerating and standing for 24 h-48 h, filtering, concentrating until the relative density is 1.10-1.20 at 20 ℃, and recovering the ethanol to obtain astragalus extract;

(4) cleaning Hirudo, oven drying, pulverizing, and sieving with 120 mesh sieve to obtain Hirudo powder;

(5) and (3) fully and uniformly mixing the extract prepared in the step (2) and the extract prepared in the step (3) and the leech powder prepared in the step (4), drying, crushing, sieving by a 120-mesh sieve, adding a pharmaceutically acceptable excipient, and preparing the tablet.

Example 4

The preparation method of the Tongguan capsule for preparing the medicine for treating hypertension and heart failure comprises the following steps:

(1) weighing the raw material medicines of astragalus, salvia miltiorrhiza and leech respectively, wherein the weight percentages of the raw material medicines are as follows: astragalus root: red sage root: leech 55%: 40%: 5 percent;

(2) mixing the astragalus and the salvia, adding 10 times of water each time, decocting for three times, boiling for 1h, 45min and 45min each time respectively, filtering through 60-mesh filter cloth while hot, combining the three filtrates, concentrating until the water content is less than or equal to 8%, cooling to room temperature, slowly adding ethanol with the volume ratio of 60%, refrigerating and standing for 24 h-48 h, filtering, concentrating, and recovering the ethanol to obtain an astragalus-salvia concentrated solution;

(3) adding 10 times of water into Hirudo, decocting for three times, boiling for 1 hr, 45min and 45min, filtering with 60 mesh filter cloth, mixing filtrates, and concentrating the filtrate under reduced pressure to obtain Hirudo concentrated solution;

(4) and (3) uniformly mixing the concentrated solutions prepared in the step (2) and the step (3), adding pharmaceutically acceptable excipient, and preparing the oral liquid or the dropping pill.

The effect of the Tongguan capsule in treating hypertensive heart failure is demonstrated by animal experiments, wherein the Tongguan capsule is prepared according to the preparation method shown in example 1.

Animal molding

SPF SD male rats 90, with weight 230-280g, were randomly divided into 10 control groups, 30 model groups, 25 positive (metoprolol) control groups and 25 open crown capsule groups. Rats were recorded for survival given the corresponding drug intervention, where:

control group (CON): injecting normal saline into abdominal cavity once a day, and irrigating stomach with distilled water for one week continuously;

model group (ISO): performing subcutaneous injection of isoproterenol 10mg/kg once a day, and performing intragastric administration with distilled water for one week continuously;

positive control group (MET): performing intraperitoneal injection of 10mg/kg of isoproterenol and 30mg/kg of metoprolol once a day for one week continuously;

coronary capsule group (TG): the isoproterenol is injected into the abdominal cavity at the dose of 10mg/kg twice a day, and the capsule for treating coronary disease is injected into the stomach at the dose of 0.5g/kg, and is continuously used for one week.

In the animal molding process, the survival of the mice is shown in figure 1, and it can be seen from the figure that the survival rate of the CON group is 100%, the survival rate of the ISO group is 53.66%, the survival rate of the MET group is 75%, and the survival rate of the TG group is 78.05%. The survival rate of the ISO group is obviously reduced compared with that of the control group (P is less than 0.001), the survival rate of the TG group is obviously improved compared with that of the ISO group (P is less than 0.05), and the survival rate of the MET group has an improvement trend, but has no statistical significance compared with that of the ISO group.

Comparison of cardiac function parameters of rats in groups II and III

1. Test method

After one week of administration, using an ultra-high resolution small animal ultrasound imaging system (Vevo2100, Visual Sonies), anesthetizing with tribromoethanol, t-amyl alcohol, atropine, anesthetizing the rat, placing on a constant temperature pad at 37 ℃, connecting the ends of the four limbs of the rat with 4 electrodes through a conductive paste, and fixing with an adhesive tape. Removing rat chest hair with a razor and depilatory cream, coating an ultrasonic coupling agent on the rat chest region, performing echocardiography on the rat by using an RMV707B type high-frequency ultrasonic probe, placing the probe on the left chest of the rat, moving the section to a left ventricle short axis section at the papillary muscle level after obtaining a satisfactory sternal side left ventricle long axis two-dimensional image, enabling the papillary muscle to be located in the four o' clock direction, recording ventricular wall motion through M-type ultrasonic, analyzing cardiac structure function, and calculating left ventricle Ejection Fraction (EF) and left ventricle short axis shortening rate (FS).

Hypertensive heart failure causes a decline in left ventricular contractile function of the heart, and EF and FS are the first indicators for determining left ventricular contractile ability and therefore can be used to assess the severity of heart failure.

2. Test results

The echocardiogram of each group of rats is shown in fig. 2, the EF and FS measurement results are shown in fig. 3 and fig. 4, and the graphs show that the EF value of the ISO group is obviously reduced (p is less than 0.05) compared with the CON group, the EF value of the TG group is obviously improved (p is less than 0.05) compared with the ISO group, and the MET group has an increasing trend but has no statistical significance compared with the ISO group.

According to experimental results, the EF and FS of rats can be obviously increased after the TG group is treated, and the heart failure can be effectively improved.

Weighing heart, lung and liver and measuring length of tibia

1. Test method

Weighing the rat, after anesthesia, taking out the heart, lung and liver of the rat, separating the left ventricle and weighing; and taking out the left tibia, measuring the length of the tibia by using a vernier caliper, and calculating the heart-body ratio, the heart-tibia ratio, the lung-body ratio, the lung-diameter ratio and the like.

2. Test results

The lung-to-shank ratio and heart-to-shank ratio measurements of the rats in each group are shown in fig. 5 and fig. 6, respectively, the lung-to-shank ratio and the heart-to-shank ratio CON in the ISO group are obviously increased (the lung-to-shank ratio P is less than 0.01 and the heart-to-shank ratio P is less than 0.001), and the lung-to-shank ratio is obviously reduced (P is less than 0.01) and the heart-to-shank ratio is obviously reduced (P is less than 0.05) in the ISO group compared with the ISO group after MET and TG treatments.

The measurement results of the lung body ratio and the heart body ratio of rats in each group are respectively shown in fig. 7 and fig. 8, the lung body ratio of the ISO group is obviously increased compared with the CON group (the lung body ratio P is less than 0.01), and the lung body ratio of the ISO group is obviously reduced compared with the lung body ratio of the ISO group (P is less than 0.01) after MET and TG treatment. In ISO group, the CON group was significantly elevated (heart-to-body ratio P < 0.001), and after MET and TG treatments, heart was reduced compared with ISO group, but it was not statistically significant.

Fourth, evaluation of myocardial hypertrophy and fibrosis

1. Test method

After the heart specimen was taken out, it was fixed with 4% paraformaldehyde solution for 24 hours, and then dehydrated and embedded in paraffin by a conventional method. The cardiomyocytes were subjected to HE staining and sirius red staining to observe cardiomyocyte hypertrophy and changes in the extracellular matrix proteins of the cardiomyocytes.

2. Test results

As shown in fig. 9 and fig. 10, the general appearance of the heart, HE staining of cardiomyocytes, sirius red staining patterns were shown for different groups after one week of administration, respectively, as observed from HE stained sections of cardiomyocytes: the myocardial cells of the MET group and the TG group are orderly and compact in arrangement and complete in shape; the arrangement of the myocardial cells of the ISO group and the CON group is disordered and loose, the volume of the myocardial cells is obviously increased, and the shape is irregular.

Fig. 11 and 12 are a cross-sectional area graph and a fibrosis ratio graph of myocardial cells of different groups, respectively, and the results show that: compared with the CON group, the heart of the ISO group is obviously increased, the transverse cutting area of the myocardial cells is obviously increased (p is less than 0.001), and the myocardial cell hypertrophy is obviously improved (p is less than 0.001) after the MET and TG are treated; tianlang scarlet staining shows that ISO causes cardiac tissue fibrosis (p < 0.01), and MET and TG interventions can obviously improve cardiac fibrosis (MET p < 0.01 and TG p < 0.01).

Fifth, Western Blot for detecting cardiac tissue protein expression

1. Test method

Ventricular tissue was homogenized and frozen, and expression of the pathway proteins CaMKII δ, HDAC4, MEF2, MAPKs and GATA4 was assessed by SDS polyacrylamide gel electrophoresis, membrane transfer (PVDF membrane), blocking, hybridization, chemiluminescence visualization.

2. Test results

As can be seen from FIGS. 13-17, ISO caused a significant increase in phosphorylation levels of the pathway proteins MAPKs (ERK, JNK and p38) (p < 0.05), and MET and TG treatment inhibited phosphorylation activation of MAPKs (p < 0.05). ISO caused increased CaMKII delta expression, MET and TG treatment reduced ISO caused CaMKII delta upregulation (p < 0.05).

As can be seen from FIGS. 18-20, ISO caused the transcriptional inhibitor HDAC4 to be transferred from nucleus to cytoplasm, the expression of cytoplasmic HDAC4 was up-regulated (p < 0.01), and both MET and TG treatments could significantly reduce the level of cytoplasmic HDAC4 (MET p < 0.01; TG p < 0.05). ISO caused the transcription factors MEF2 and GATA4 to be up-regulated (p < 0.05), MET and TG can reduce their expression (p < 0.05).

Therefore, the characterization of the above experiment and data results shows that the coronary circulation promoting capsule has a good effect of improving hypertensive heart failure. Moreover, the medicine for treating hypertension and heart failure has the advantages of less medicine taste, definite and reliable curative effect and definite medicine effect components.

What has been described above are merely some embodiments of the present invention. It will be apparent to those skilled in the art that various changes and modifications can be made without departing from the inventive concept thereof, and these changes and modifications can be made without departing from the spirit and scope of the invention.

Claims (7)

1. The application of the Tongguan capsule in preparing the medicine for treating hypertension and heart failure,

the active ingredients of the Tongguan capsule comprise the following raw materials in percentage by weight: 40-60% of astragalus membranaceus, 20-50% of salvia miltiorrhiza and 5-25% of leech.

2. The use according to claim 1, wherein the medicament consists of the active ingredient in the Tongguan capsule and pharmaceutically acceptable excipients.

3. The use of claim 2, wherein the medicament is a capsule, soft capsule, granule, tablet, drop pill or oral liquid.

4. The use according to any one of claims 1 to 3, wherein the medicament is prepared by the following preparation method:

(1) weighing Saviae Miltiorrhizae radix and Hirudo according to a certain proportion, cleaning, oven drying, pulverizing, and sieving with 120 mesh sieve to obtain Saviae Miltiorrhizae radix-Hirudo powder;

(2) weighing radix astragali according to a ratio, adding 10 times of water each time, decocting for three times, boiling for 1h, 45min and 45min each time, filtering through 60-mesh filter cloth while hot, combining the three filtrates, concentrating until the water content is less than or equal to 8%, cooling to room temperature, slowly adding 60% ethanol by volume, refrigerating and standing for 24 h-48 h, filtering, concentrating until the relative density is 1.10-1.20 at 20 ℃, and recovering ethanol to obtain radix astragali extract;

(3) and (3) fully and uniformly mixing the salvia-leech powder prepared in the step (1) and the astragalus extract prepared in the step (2), drying, crushing, sieving by a 120-mesh sieve, adding a pharmaceutically acceptable excipient, and preparing into capsules, granules, tablets or soft capsules.

5. The use according to any one of claims 1 to 3, wherein the medicament is prepared by the following preparation method:

(1) weighing the astragalus and the salvia miltiorrhiza according to the proportion, adding 10 times of water each time, decocting for three times, boiling for 1h, 45min and 45min each time respectively, filtering through 60-mesh filter cloth while the mixture is hot, combining the three filtrates, concentrating until the water content is less than or equal to 8%, cooling to room temperature, slowly adding 60% ethanol by volume, refrigerating and standing for 24 h-48 h, filtering, concentrating until the relative density is 1.10-1.20 at 20 ℃, and recovering the ethanol to obtain astragalus-salvia miltiorrhiza extract;

(2) weighing leeches according to the proportion, cleaning, drying, crushing, and sieving with a 120-mesh sieve to obtain leech powder;

(3) and (3) fully and uniformly mixing the extract prepared in the step (1) and the leech powder prepared in the step (2), drying, crushing, sieving by a 120-mesh sieve, and adding a pharmaceutically acceptable excipient to prepare capsules, granules, tablets or soft capsules.

6. The use according to any one of claims 1 to 3, wherein the medicament is prepared by the following preparation method:

(1) weighing salvia miltiorrhiza according to a ratio, adding 7 times of ethanol, heating and refluxing for 1.5h, filtering, adding 50% filtrate of ethanol into dregs of a decoction, heating and refluxing for 1h, filtering, combining filtrates, concentrating to a relative density of 1.10-1.20 at 20 ℃, and recovering ethanol to obtain salvia miltiorrhiza extract;

(2) weighing astragalus membranaceus according to a ratio, adding the astragalus membranaceus into the salvia miltiorrhiza dregs, adding 10 times of water each time, decocting for three times, boiling for 1 hour, 45 minutes and 45 minutes respectively each time, filtering through 60-mesh filter cloth while the decoction is hot, combining filtrate obtained in the three times, concentrating the filtrate under reduced pressure until the water content is less than or equal to 8%, cooling to room temperature, adding ethanol with the volume ratio of 60%, refrigerating and standing for 24 hours to 48 hours, filtering, concentrating until the relative density is 1.10-1.20 at 20 ℃, and recovering the ethanol to obtain an astragalus membranaceus extract;

(3) weighing leeches according to the proportion, cleaning, drying, crushing, and sieving with a 120-mesh sieve to obtain leech powder;

(4) and (3) fully and uniformly mixing the extract prepared in the steps (1) and (2) and the leech powder prepared in the step (3), drying, crushing, sieving by a 120-mesh sieve, and adding a pharmaceutically acceptable excipient to prepare capsules, granules, tablets or soft capsules.

7. The use according to any one of claims 1 to 3, wherein the medicament is prepared by the following preparation method:

(1) weighing the astragalus and the salvia according to the proportion, adding 10 times of water each time, decocting for three times, boiling for 1h, 45min and 45min each time respectively, filtering through 60-mesh filter cloth while the mixture is hot, combining the three filtrates, concentrating until the water content is less than or equal to 8%, cooling to room temperature, slowly adding ethanol with the volume ratio of 60%, refrigerating and standing for 24 h-48 h, filtering, concentrating, and recovering the ethanol to obtain astragalus-salvia concentrated solution;

(2) weighing Hirudo according to a certain proportion, adding 10 times of water, decocting for three times, boiling for 1 hr, 45min and 45min each time, filtering with 60 mesh filter cloth while hot, mixing filtrates, and concentrating the filtrate under reduced pressure to obtain Hirudo concentrated solution;

(3) and (3) uniformly mixing the concentrated solutions prepared in the step (1) and the step (2), adding pharmaceutically acceptable excipient, and preparing the oral liquid or the dropping pill.

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