阅读说明：本技术 硫酸舒欣啶在制备抗心衰产品中的应用 (Application of sulcardine sulfate in preparation of anti-heart failure products ) 是由 陈民利 马全鑫 白东鲁 胡有洪 张冠亚 金昌明 吕建东 于 2019-09-17 设计创作，主要内容包括：本发明公开了硫酸舒欣啶在制备抗心衰产品中的应用,属于药物技术领域。本发明能降低左室室间隔厚度,抑制心脏负荷增大所致的心肌肥大和心室重构；通过改善心电生理,纠正ST段压低和QT间期延长,调节心律,抗心肌梗死；具有降低由心衰所引发的心脏质量增加,降低血浆中NT-proBNP水平的作用；具有改善心室重构和心肌肥大所致的病理学变化如细胞核增大,排列紊乱,血管结构变化,外膜结缔组织增生和降低胶原沉积的药理作用,并显著降低左室重构引发心衰的死亡率。该化合物式可用已知方法合成,制备工艺成熟,能够提供可靠的化合物来源,是一种理想的抗心衰化合物,可应用于制备预防、缓解和治疗心衰的产品。(The invention discloses an application of sulcardine sulfate in preparing an anti-heart-failure product, belonging to the technical field of medicines. The invention can reduce the left ventricular septum thickness, and inhibit myocardial hypertrophy and ventricular remodeling caused by cardiac load increase; by improving the electrocardio physiology, correcting ST-segment depression and QT interval prolongation, regulating the heart rhythm and resisting myocardial infarction; has the effects of reducing heart mass increase caused by heart failure and reducing the level of NT-proBNP in blood plasma; has the pharmacological effects of improving pathological changes caused by ventricular remodeling and myocardial hypertrophy, such as nuclear enlargement, disorganization, vascular structure change, adventitial connective tissue hyperplasia and collagen deposition reduction, and remarkably reducing the mortality of heart failure caused by left ventricular remodeling. The compound formula can be synthesized by a known method, has mature preparation process, can provide a reliable compound source, is an ideal anti-heart failure compound, and can be applied to preparation of products for preventing, relieving and treating heart failure.)

1. Application of sulcardine sulfate in preparing anti-heart failure products.

2. The use of claim 1, wherein sulcardine sulfate is used as a basic parent nucleus, and the use of glycosides compounds generated by combination of saccharides and saccharides with saccharide structures is used in the anti-heart failure product.

3. The use of claim 1, wherein the heart failure is congestive heart failure due to cardiomyopathy or cardiac overload.

4. The use according to claim 1, wherein the anti-heart failure is improvement of myocardial hypertrophy and ventricular remodeling by sulcardine sulfate reducing left ventricular septal thickness and left ventricular posterior wall thickness.

5. The use as claimed in claim 1, wherein the anti-heart failure is by sulcardine sulfate to improve ST depression and QT interval prolongation, to regulate heart rhythm, or to resist myocardial infarction.

6. The use according to claim 1, wherein the anti-heart failure is a decrease in the level of NT-proBNP in the plasma by decreasing the increase in heart mass due to heart failure with sulcardine sulfate.

7. The use of claim 1, wherein the anti-heart failure is the improvement of nuclear enlargement caused by heart failure, disorganization of myocardial cells, reduction of collagen deposition and reduction of heart failure mortality by sulcardine sulfate.

8. The use according to claim 1, wherein the product is any one of a pharmaceutical product, a food product and a health product.

9. A pharmaceutical composition for resisting heart failure, which comprises an effective amount of sulcardine sulfate or a pharmaceutically acceptable salt, hydrate thereof and a pharmaceutically acceptable carrier, adjuvant or combination thereof.

10. The pharmaceutical composition of claim 9, wherein the pharmaceutical composition is in the form of any one of powder, granule, tablet and capsule.

Technical Field

The invention belongs to the technical field of medicines, and relates to application of sulcardine sulfate in preparation of heart failure resistant products.

Background

Heart Failure (HF), abbreviated as heart failure, is not a single disease but a clinical syndrome of cardiac insufficiency caused by various heart diseases, and is also the ultimate fate of most cardiovascular diseases. HF and arrhythmias are often associated with clinical use. Most antiarrhythmic drugs have the characteristics of inhibiting heart conduction, inhibiting myocardial contraction and the like, so that the clinical treatment of the drugs for treating heart failure combined arrhythmia is difficult^[1]. Therefore, it is important to find a drug which is effective on arrhythmia and can be safely used by heart failure patients or even obtain benefits.

Chronic cardiac dysfunction diseases caused by multiple factors of HF, dyspnea, fatigue, impaired exercise tolerance and fluid retention are common diseases of HF, diseases such as hypertension, coronary heart disease, cardiomyopathy, arrhythmia, pericardial disease and the like are common causes of heart failure, wherein the hypertension and the coronary heart disease are the main causes of HF at present. Congestive heart failure caused by hypertension generally occurs and progresses from structural heart disease to the appearance of heart failure symptoms to refractory end-stage heart failure. At present, HF treatment strategies mainly comprise beta receptor blockers, angiotensin 2 convertase inhibitors and other antihypertensive drugs, while arrhythmia treatment drugs mainly comprise ion channel inhibitors, and no drug which has the effect of preventing and treating HF and arrhythmia is available at present.

The sulcardine sulfate is an inhibitor of sodium, calcium and potassium multi-ion channels, is a novel structural type drug with anti-arrhythmia activity, the current research aiming at the sulcardine sulfate mainly focuses on the aspect of treating arrhythmia, and the research finds that the influence of the compound on heart failure is not reported so far. Based on the antagonism of sulcardine sulfate on calcium ion channels, the inventor thinks that sulcardine sulfate may also have a certain prevention and treatment effect on HF.

Disclosure of Invention

The purpose of the invention is as follows: the invention aims to solve the technical problem of providing the application of sulcardine sulfate in preparing anti-heart-failure products aiming at the defects of the prior art.

In order to solve the technical problems, the invention discloses application of sulcardine sulfate in preparing a heart failure resistant product.

Wherein the chemical structure of sulcardine sulfate is shown as formula I,

the sulcardine sulfate can be prepared by known methods, refer to patent CN 1299813N-substituted benzyl or phenyl aromatic sulfonamide compound and application thereof.

The application of a glycoside compound generated by taking sulcardine sulfate as a basic parent nucleus, saccharides and combination with saccharide structures in anti-heart-failure products.

Wherein the heart failure is congestive heart failure caused by myocardial pathology or cardiac overload, and the pathology is characterized by myocardial hypertrophy and left ventricular remodeling.

Wherein, the heart failure resistance is effective prevention, alleviation and treatment of heart failure.

The invention utilizes the principle that the load after increasing the left ventricular pressure enables the myocardium to generate compensatory thickening and fibrosis so as to cause the heart failure, adopts a congestive heart failure animal model established by rat left ventricular pressure overload caused by aortic arch constriction, proves the prevention and treatment effect of the sulcardine sulfate on HF, and finds that the sulcardine sulfate has certain improvement effect on the myocardial collagen deposition and ventricular remodeling of the rat model with early heart failure.

The sulcardine sulfate can reduce the left ventricular septum thickness and the left ventricular posterior wall thickness, and inhibit myocardial hypertrophy and ventricular remodeling caused by cardiac load increase; by improving the electrocardio physiology, correcting ST-segment depression and QT interval prolongation, regulating the heart rhythm and resisting myocardial infarction; has the effects of reducing the increase of heart mass caused by the increase of heart load and reducing the level of NT-proBNP in blood plasma; has pharmacological effects of improving pathological changes of myocardial hypertrophy and ventricular remodeling such as nuclear enlargement, disorganization, loss of basic structure of blood vessel, proliferation of connective tissue of adventitia and reduction of collagen deposition, and remarkably reduces mortality. Therefore, the preparation method can be applied to preparing products for preventing, relieving and treating heart failure.

Wherein the product is any one of medicine, food and health care product.

Wherein the heart failure is congestive heart failure caused by myocardial lesion or cardiac overload.

A pharmaceutical composition for resisting heart failure, which comprises an effective amount of sulcardine sulfate or a pharmaceutically acceptable salt, hydrate thereof and a pharmaceutically acceptable carrier, adjuvant or combination thereof.

The pharmaceutical composition is prepared by combining sulcardine sulfate or pharmaceutically acceptable salts and hydrates thereof with other substances in any proportion.

Wherein the dosage form of the pharmaceutical composition is any one of powder, granules, tablets and capsules.

The preparation method of the powder comprises the steps of crushing the ingredients in the pharmaceutical composition, sieving, mixing, dividing the dosage and packaging.

The preparation method of the granules comprises the steps of crushing, sieving, mixing, granulating, drying, grading and internally packaging the ingredients in the pharmaceutical composition, wherein the known pharmaceutical excipients can be added into the granules.

The preparation method of the tablet comprises the steps of crushing the ingredients in the pharmaceutical composition, sieving, blending, granulating, drying, granulating, mixing, tabletting, coating and subpackaging, and the tablet is obtained, wherein the known pharmaceutical excipients can be added into the pharmaceutical excipients.

The preparation method of the capsule comprises the steps of weighing the ingredients in the pharmaceutical composition, crushing, sieving, granulating, drying, granulating, totally mixing, filling and subpackaging, and the capsule is obtained, wherein the known pharmaceutical excipients can be added into the pharmaceutical excipients.

Has the advantages that: compared with the prior art, the invention has the following advantages:

(1) the sulcardine sulfate researched by the invention can be synthesized by a known method, has a mature preparation process, can provide a reliable compound source, and has the advantages of reliable material source, low production cost, novel action mechanism, good market prospect and the like.

(2) The invention has the advantages of reducing the left ventricular septal thickness, inhibiting myocardial hypertrophy and ventricular remodeling caused by hypertension; by improving the electrocardio physiology, correcting ST-segment depression and QT interval prolongation, regulating the heart rhythm and resisting myocardial infarction; has the effects of reducing heart mass increase caused by heart failure and reducing the level of NT-proBNP in blood plasma; has pharmacological effects of improving pathological changes caused by heart failure such as nuclear enlargement, disorganization, loss of basic structure of blood vessel, proliferation of connective tissue of adventitia and reduction of collagen deposition, and remarkably reducing death rate of heart failure. Can be used for preparing products for preventing, relieving and treating heart failure.

Drawings

FIG. 1 is a graph of the effect on left ventricular septal thickness (IVS) of rats in a heart failure model, annotated as compared to the sham operated group,^#P<0.05，^##P<0.01; compared with the model control group,^*P<0.05,^**P<0.01。

FIG. 2 is a graph of the effect on the posterior wall thickness of the Left Ventricle (LVPW) in rats in the model of heart failure 2 weeks after administration, annotated as compared to the sham-operated group,^#P<0.05，^##P<0.01; compared with the model control group,^*P<0.05,^**P<0.01。

FIG. 3 is a graph of the effect on heart failure model rat electrocardiograms 2 weeks after administration, annotated as compared to the sham-operated group,^#P<0.05，^##P<0.01; compared with the model control group,^*P<0.05,^**P<0.01。

FIG. 4 is a graph of the effect on heart failure model rat heart coefficients 2 weeks after administration, annotated as compared to sham,^#P<0.05，^##P<0.01; compared with the model control group,^*P<0.05,^**P<0.01。

FIG. 5 is the effect on the plasma NT-proBNP content of rats in the model of heart failure 2 weeks after administration, annotated as compared to the sham-operated group,^#P<0.05，^##P<0.01; compared with the model control group,^*P<0.05,^**P<0.01。

FIG. 6-1 shows HE staining results of myocardial tissues in the sham-operated group, 10 XHE staining.

FIG. 6-2 shows HE staining results of myocardial tissues in the sham-operated group, 200 XHE staining.

FIGS. 6-3 show HE staining results of myocardial tissues of the model control group, 10 XHE staining.

FIGS. 6-4 show HE staining results of myocardial tissues of the model control group, 200 XHE staining.

FIGS. 6-5 show HE staining results, 10 XHE staining, of myocardial tissue in the low dose group.

FIGS. 6-6 show HE staining results, 200 XHE staining, of myocardial tissue in the low dose group.

FIGS. 6-7 show HE staining results, 10 XHE staining, of myocardial tissues in the medium dose groups.

FIGS. 6-8 show HE staining results, 200 XHE staining, of myocardial tissue in the medium dose group.

FIGS. 6-9 show HE staining results, 10 XHE staining, of myocardial tissue in the high dose group.

FIGS. 6-10 show HE staining results, 200 XHE staining, of myocardial tissue in the high dose groups.

FIGS. 6 to 11 show HE staining results of myocardial tissues of the positive control group, 10 XHE staining.

FIGS. 6 to 12 show HE staining results of myocardial tissues of the positive control group, 200 XHE staining.

FIG. 7 is a graph of the effect on myocardial fibrosis score in rats in a heart failure model.

FIG. 8-1 shows Masson staining of myocardial tissue in the sham-operated group, 10 XMasson staining.

FIG. 8-2 shows Masson staining of myocardial tissue in the sham-operated group, 100 XMasson staining.

FIGS. 8-3 show Masson staining of myocardial tissue in the model control group and 10 XMasson staining.

FIGS. 8-4 show Masson staining of myocardial tissue in the model control group and 100 XMasson staining.

FIGS. 8-5 show Masson staining of myocardial tissue in the low dose groups, 10 XMasson staining.

FIGS. 8-6 show Masson staining of myocardial tissue in the low dose groups, 100 XMasson staining.

FIGS. 8-7 show Masson staining of myocardial tissues in the medium dose groups, 10 XMasson staining.

FIGS. 8-8 show Masson staining of myocardial tissues in the medium dose groups, 100 XMasson staining.

FIGS. 8-9 show Masson staining of myocardial tissue in the high dose groups, 10 XMasson staining.

FIGS. 8-10 show Masson staining of myocardial tissue in the high dose groups, 100 XMasson staining.

FIGS. 8-11 show Masson staining, 10 XMasson staining, of myocardial tissue in the positive control group.

FIGS. 8-12 show Masson staining, 100 XMasson staining of myocardial tissue in the positive control group.

Detailed Description

The invention will be better understood from the following examples. However, those skilled in the art will readily appreciate that the description of the embodiments is only for illustrating the present invention and should not be taken as limiting the invention as detailed in the claims.

Example 1: effect of sulcardine sulfate on early heart failure model in rats

1. Purpose of the experiment

In the embodiment, an aortic arch stenosis (TAC) is adopted to establish an early-stage Heart Failure (HF) model of a rat, low (50mg/kg), medium (150mg/kg) and high (200mg/kg) doses of sulcardine sulfate is administered once a day for 2 weeks continuously, echocardiogram detection and electrocardiogram analysis are performed, the content of plasma NT-proBNP is measured, heart histopathological examination is performed, the deposition condition of collagen fibers in myocardial tissues is observed, and the effect of the sulcardine sulfate on the heart function of the heart failure model of the rat is observed by taking captopril tablets as positive control, so that reference is provided for the clinical application of the sulcardine sulfate.

2. Test drug

2.1 test drugs

Sulcardine Sulfate (SUL), trait: white powder, batch number: d17120401; providing a unit: yangzhou Chinese medicine industry, stock Co Ltd, storage mode: protected from light at room temperature.

2.2 Positive control

Captopril tablet (Captopril), trait: white tablets, lot number: T16L 068; purchased from Hangzhou Minsheng pharmaceutical Co., Ltd, the specification: 25mg × 100 tablets, storage mode: protected from light at room temperature.

2.3 negative control

Distilled water

3. Test animal

SPF grade SD rats 92, sex: male, body weight 170-. The source is as follows: shanghai slek laboratory animals ltd, production license number: the animal quality qualification number of SCXK (Shanghai) 2017-: 2015000549137.

4. test conditions

Rat rearing room: SPF barrier environment, temperature: 20-22 ℃, humidity: 50-70%, illumination: light and shade alternation within 12 hours (6: 00 early to 18:00 late) at 150-200 Lx; noise: <50 dB; an environment automatic control system is arranged. Application license of barrier environmental facility in animal experiment research center of Zhejiang university of traditional Chinese medicine [ SYXK (Zhe) 2013-.

Drinking water: filtering and sterilizing tap water, and placing in an autoclaved drinking bottle for free drinking.

Feed: complete nutrition rat pellet feed, Co60 irradiation sterilization.

The breeding mode is as follows: free diet, housed in IVC, given sufficient water and feed, rats were housed 3-4 per cage.

Throughout the experimental period, the experimental feeding and handling of rats was performed in accordance with the 3R principle used by experimental animals for humanistic care.

5. Reagent and apparatus

5.1 Primary reagents

N-terminal pro-brain natriuretic peptide (NT-proBNP) ELISA kit, batch No. 07/2018, purchased from Nanjing Biotech.

5.2 Main Instrument

SQP electronic balance, sydows scientific instruments (beijing) ltd; 7020 full-automatic biochemical analyzer, Hitachi corporation; varioskan Flash multifunctional microplate reader, Thermo corporation; small animal anesthesia respirator, summit corporation; vevo1100 small animal ultrasound imaging system, Fujifilm corporation; EMKA noninvasive physiological signal telemetry system, EMKA corporation; AP280-2 embedding machine, MICROM semi-automatic slicer, Leica company; a digital slice scanning system, a NanoZoomer digital slice scanner; hamamatsu photonics corporation.

6. Grouping and administration of drugs

6.1 groups and dosages

6.2 dose setting basis

According to the previous researchers, the effective doses of sulcardine sulfate on rat arrhythmia models are 100mg/kg, 150mg/kg and 200mg/kg (oral gavage).

6.3 route of administration

The oral gavage is carried out, 10mL/kg, and the administration route is basically consistent with that of the clinic.

7. Test method

7.1 Molding and screening

92 male SD rats were taken and after 1 week of acclimatization, 86 of them underwent aortic arch stenosis surgery: after rat is subjected to induction anesthesia by using isoflurane, the chest hair is shaved off, the rat is fixed on a 45-degree inclined fixing table, trachea intubation is carried out, then the animal is fixed on a heat preservation table, a small animal anesthesia respirator is connected externally to assist respiration, the respiration ratio is 2: 1, the tidal volume is 30mL/kg, and the respiratory frequency is 60-65 times/min. And (3) disinfecting the thoracic operation area by iodophor, cutting off a second rib on the left side of the rat, performing layer-by-layer thoracotomy, performing distraction of an incision by using a distractor, fully exposing the thymus, sequentially separating the thymus and the aortic arch, wrapping the rear wall of the aortic arch by using a No. 3-0 suture, and stretching out from between the brachiocephalic trunk and the left common carotid artery. And (3) placing a self-made L-shaped needle with the outer diameter of 0.9mm above the aortic arch in parallel, slowly taking out the L-shaped needle after ligation, closing the thoracic cavity layer by layer, and disinfecting the incision. After the operation is finished, the animal is put into the incubator for warming and nursing until the animal wakes up; another 6 rats underwent sham surgery, and the aortic arch was not narrowed in the same procedure as the sham surgery group. After operation, 100000U penicillin is injected into the muscle at 3d, and the breeding is carried out normally to 4 weeks after 3 d.

After 4 weeks, the stored animals were subjected to echocardiography, ventricular wall thickness was recorded, and serum NT-proBNP levels were measured by bleeding. And according to the heart super-examination result and the serum NT-proBNP detection result, 40 heart failure rat models with relatively close indexes are screened for a drug administration test.

7.2 grouping and administration

The method comprises the following steps of taking the left ventricular Mass (LV Mass) measured and calculated by an ultrasonic system as a main index of 40 screened TAC postoperative heart failure rat models, and dividing the model rats into 5 groups according to weight, namely a model control group, a low, medium and high dose group and a positive control group, wherein each group comprises 8 rats. The 6 rats subjected to additional sham surgery were sham operated groups, and each group was administered the corresponding drug once a day for 2 weeks (14 days).

7.3 general signs Observation

During the test period, the mental state, the fecal condition and the fur color and state of the rat are observed, the survival condition of the animal after the operation is recorded, and the survival rate is calculated.

7.4 cardiac ultrasonography and electrocardiographic monitoring

The administration is performed for 1 week and 2 weeks under anesthesia with isoflurane gas, and electrocardiogram is detected after 2-3 hours of last administration at 2 weeks. The specific method comprises the following steps: carrying out isoflurane inhalation anesthesia on a rat, connecting an electrocardio electrode after the rat is fixed and stabilized for 5min in a supine way, monitoring the change of an electrocardiogram of the rat for 8-10 min, then measuring the left ventricular end diastolic chamber interval thickness (IVS, d), the left ventricular end systolic chamber interval thickness (IVS, s), the left ventricular end diastolic wall thickness (LVPW, d) and the left ventricular end systolic wall thickness (LVPW, s) by using a high-resolution small animal ultrasonic imaging system, measuring each index on 2-3 cardiac cycles and taking the average value of the indexes.

7.5 Biochemical index determination

After the experiment is finished, the content of the plasma N-terminal brain natriuretic peptide (NT-proBNP) is detected by an ELISA method.

7.6 histopathological Observation of the Heart

(1) Rat heart weight and heart index calculation

After blood collection of rats, the thoracic cavity is opened to pick up the heart, other tissues such as the auricle, the ascending aorta and the like are removed, the wet weight is weighed by an electronic balance, and the heart index is calculated.

(2) Masson staining observation of rat cardiac tissue

And (3) fixing the removed heart in a 10% neutral formaldehyde solution for 2 days, dehydrating, transparentizing, waxing and embedding. After wax block formation, 4 μm sections were cut with a microtome for Masson staining, as specified in the kit instructions. Then, a Nano zoom 2.0RS digital slice scanner is used for scanning and shooting tissue slices, the number and the area of collagen fiber deposition areas are observed, the myocardial collagen deposition degree is divided into 0-4 grades, integral scoring is firstly carried out, then, 3 parts with collagen deposition are taken from each animal, the characteristics of collagen distribution are observed, scoring is carried out, and the scoring standard is as follows:

0 minute: the collagen fibers are basically free of blue color, and the occasional blue collagen fibers are scattered in a dotted manner;

1 minute: the myocardial fibers are arranged orderly, and a small amount of blue collagen fibers are distributed in the myocardial interstitium in a dotted manner;

and 2, dividing: more blue collagen fibers are scattered in a dotted manner, and parts of the blue collagen fibers are connected into a belt shape;

and 3, dividing: the myocardial cells are loose and are divided by obviously proliferated and thickened collagen fibers to present 'mesh-shaped' change

And 4, dividing: a great deal of blue collagen fibers are distributed in a sheet shape, and the collagen fibers between the cardiac muscle matrixes are excessively deposited.

The scores of the 3 photographs were added and the total score for each animal was counted.

8. Observation index

8.1 general index

Survival rate, weight, behavioral activity and dietary status;

8.2 index of cardiac function

IVS, d, IVS, s, LVPW, d, LVPW, s, etc.;

8.3 Electrocardiogram analysis

Heart Rate (HR), ST elevation (STE), QT interval and T-wave amplitude, incidence of 5min arrhythmia;

8.4 Biochemical index

NT-proBNP；

8.5 histopathological Observation

HE staining and Masson staining (observation of collagen fiber deposition in myocardial tissue).

9. Data processing

Statistical analysis was performed using SPSS 22.0 software, all data averaged. + -. standard errorAnd (3) expressing that the measurement data adopts ANOVA analysis of variance to evaluate the test result, comparing every two data by adopting LSD test, and keeping the numerical value of the statistical result at 2 bits after decimal. Data analysis processing, each group allows to reject 1 abnormal data (more thanAbnormal ranges and experimental conditions).

10. Test results

10.1 Effect on general signs of rats in Heart failure model

During the administration period, the rest groups have no death except 1 death at 2 days after the administration of the model control group and 1 death at 9 days after the administration of the positive control group, and no obvious abnormality is seen in diet, weight gain and behavior activities of the test rats in each group.

After 2 weeks of administration, 2 electrocardiographic rhythms of 7 surviving rats of the model control group are abnormal by continuous 5min electrocardiographic detection, wherein 1 arrhythmia, partial ventricular extrasystole and 1P wave are inverted; only 1 ecg rhythm abnormality (P-wave inversion) survived 7 of the positive control group; the heart rate of rats in the high, medium and low dose groups was not abnormal, and the heart rate was not abnormal in the sham operation group.

TABLE 1 Effect on the survival Rate and the electrocardiographic rhythm of rats in the model of cardiac failure

10.2 Effect on Heart Structure of rat model for Heart failure

10.2.1 Effect on ventricular decay model rat left ventricular septal thickness (IVS)

As can be seen from table 2 (fig. 1), compared to the sham group, both systolic ivs(s) and diastolic ivs (d) in rats were significantly increased in thickness before and at 1 and 2 weeks after administration (P <0.01, P < 0.05); when compared with a model control group, the IVS of each administration group has no significant change in the systolic period and the diastolic period when the administration is carried out for 1 week (P is more than 0.05); when the medicine is administrated for 2 weeks, the systolic phase IVS and the diastolic phase IVS of the low-dose group, the middle-dose group and the high-dose group are reduced to different degrees, wherein the systolic phase IVS of the high-dose group and the middle-dose group is reduced remarkably (P < 0.05); the positive control group also had different degrees of reduction in systolic and diastolic IVS, with a significant reduction in diastolic IVS (P < 0.01).

TABLE 2 Effect on left ventricular septal thickness (IVS) of rats in the model of heart failure

Note: compared with the group of the pseudo-operation,^#P<0.05，^##P<0.01; compared with the model control group,^*P<0.05,^**P<0.01。

10.2.2 Effect on left ventricular posterior wall thickness (LVPW) of rats in heart failure model

As can be seen from table 3 and fig. 2, both systolic(s) and diastolic (d) LVPW increased significantly before and 2 weeks after dosing in the model control group (P <0.05, P <0.01) compared to the sham group; compared with the model control group, the systolic LVPW of the medium and high dose groups was significantly reduced at 2 weeks of administration (P <0.05, P < 0.01).

TABLE 3 Effect on left ventricular posterior wall thickness (LVPW) of rats in models of heart failure

Note: the model control group was compared with the sham-operated group,^#P<0.05，^##P<0.01; each administration group was compared with the model control group,^*P<0.05,^**P<0.01。

10.3 Effect on Heart failure model rat Electrocardiogram

As can be seen from table 4 and fig. 3, compared to the sham group, the QT interval time was significantly prolonged in the model control group (P <0.05, P <0.01), and the ST elevation value (STE) was significantly depressed (P < 0.01); compared with a model control group, the low, medium and high dose groups have obvious improvement on the STE depression degree (P <0.05, P <0.01), and the medium and high dose groups have obvious improvement on the QT interval time prolongation (P <0.05, P < 0.01).

TABLE 4 Effect on Heart failure model rat electrocardiograms

Note: compared with the group of the pseudo-operation,^#P<0.05，^##P<0.01; compared with the model control group,^*P<0.05,^**P<0.01。

10.4 Effect on Heart quality and coefficients of Heart failure model rats

10.4.1 Effect on Heart quality in rat model of Heart failure

As can be seen from Table 5, compared with the sham operation group, the heart mass measured by ultrasound in the model control group and the heart mass measured by dissection are both significantly increased (P < 0.01); compared with the model control group, after administration for 2 weeks, the heart quality is measured by ultrasonic, the heart quality measured by ultrasonic in the low, medium and high dose groups has descending tendency of different degrees, and the high dose group has statistical difference (P is less than 0.05); the results of anatomical weighing of the heart showed different reductions in the low, medium and positive controls, with the high dose group being the most significant (P > 0.05).

TABLE 5 Effect on Heart quality of rats in Heart failure model

Note: compared with the group of the pseudo-operation,^#P<0.05，^##P<0.01; compared with the model control group,^*P<0.05,^**P<0.01。

10.4.2 Effect on Heart coefficients of rats in Heart failure model

As can be seen from table 6 and fig. 4, compared with the sham group, the heart coefficients of the model control group, the ultrasound measurement and the anatomical weighing were both significantly increased (P < 0.01); compared with a model control group, after administration for 2 weeks, the anatomical weighing method by an ultrasonic measurement and calculation method shows that the heart coefficients of the medium and high dose groups are remarkably reduced (P <0.05, P <0.01), the heart coefficients of the positive control group have a reduction trend, but the difference is not remarkable (P > 0.05).

TABLE 6 Effect on heart failure model rat Heart coefficients

Note: compared with the group of the pseudo-operation,^#P<0.05，^##P<0.01; compared with the model control group,^*P<0.05,^**P<0.01。

10.5 Effect on Biochemical indicators of rats in Heart failure model

As can be seen from Table 7 and FIG. 5, the plasma NT-proBNP content was significantly increased (P <0.01) in the model control group at 2 weeks compared with the sham-operated group, and significantly decreased (P <0.05, P <0.01) in the medium/high dose group and the positive control group, and also decreased in the low dose group (P >0.05) but not significantly different (P <0.01) in the plasma NT-proBNP content in the middle/high dose group and the positive control group compared with the model control group.

TABLE 7 Effect on Biochemical indices of blood of rats in heart failure model

Note: compared with the group of the pseudo-operation,^#P<0.05，^##P<0.01; compared with the model control group,^*P<0.05,^**P<0.01。

10.6 pathological observation results on myocardial tissue of rat model of heart failure

10.6.1 HE staining of myocardial tissue in rats in Heart failure model

As can be seen from the results of HE staining (FIGS. 6-1 to 6-12), the cardiomyocytes in the sham-operated group were aligned, the nuclei were ovate, the staining was light and did not cause deep staining, no cardiomyocyte hypertrophy was observed, and no inflammatory cell infiltration was observed in the stroma (FIGS. 6-1 and 6-2); the cells of the model control group are loosely arranged, and can be seen to be broken, the cardiac fibers are thickened, the intercellular substance is in edema, inflammatory cell infiltration is caused, and bleeding is caused (see figures 6-3 and 6-4); compared with a model control group, the shapes of the myocardial cells of the high-dose group, the medium-dose group and the low-dose group are improved in different degrees, the structures are relatively complete, and the arrangement is relatively neat (see fig. 6-5 to fig. 6-10), wherein the low-dose group can see that the myocardial cells are slightly loose in arrangement, interstitial edema exists in cells, inflammatory cell infiltration exists (see fig. 6-5 and 6-6), the medium-dose group rarely has broken cells, the myocardial fibers are slightly thickened, a small amount of inflammatory cell infiltration exists, bleeding is less (see fig. 6-7 and 6-8), the myocardial cells of the high-dose group are neatly arranged, a few cells are broken, the myocardial fibers are slightly thickened, a small amount of inflammatory cell infiltration exists, and bleeding can be seen (see fig. 6-; the positive control group also has improved cardiomyocyte morphology (see fig. 6-11 and 6-12, which shows that the cardiomyocytes are relatively regularly arranged, few cells are broken, the myocardial fibers are slightly thickened, inflammatory cells infiltrate, and a small amount of bleeding is visible).

10.6.2 Effect on myocardial fibrosis in rats in Heart failure model

According to the Masson-stained collagen fiber deposition score results (see fig. 7), blue coloration was observed only in the peripheral coronary arteries and microvessels in the myocardial tissue of the sham-operated rats, not in the collagen fiber coloration, and the overall score was 1 or 0, while 5 out of 3 and above were overall scored in the model control group of 7 rats, 4, 2 and 3 out of 3 and above were overall scored in the low, medium and high dose group of 8 rats, and 2 out of 3 and above were overall scored in the positive control group of 7 rats.

As seen from the results of Masson staining of the myocardial tissues (FIGS. 8-1 to 8-12), no blue collagen fiber band was observed in the sham operation group, occasionally blue collagen fibers were scattered in a dotted manner, and only the periphery of the coronary artery and the microvasculature in the myocardial tissues were stained with blue color, but not with collagen fibers (FIGS. 8-1 and 8-2); compared with a sham operation group, the deposition of the interstitial collagen fibers of the myocardium of the model control group is obviously increased, so that the myocardial cells in the myocardial tissues of rats are loose and are divided by the obviously proliferated and thickened collagen fibers to be in a mesh shape, a large amount of blue collagen fibers are seriously distributed in a sheet shape, and the interstitial collagen fibers of the myocardium are deposited in a large amount (see fig. 8-3 and 8-4); the low, medium, high dose and positive control groups all had different degrees of reduction in myocardial interstitial collagen fiber deposition as compared to the model control group (see fig. 8-5 to 8-10), and the improvement was more evident in the medium and high dose groups, wherein a large range of blue collagen deposition areas were seen in fig. 8-5, the cardiomyocytes were more loose in fig. 8-6, and were segmented by collagen fibers that were significantly thickened to appear as a "mesh", a blue collagen deposition area was seen in fig. 8-7, blue collagen fibers "mesh" and dotted scattering were seen in fig. 8-8, and some were connected to form a band, a small range of blue collagen deposition areas were seen in fig. 8-9, and blue collagen fibers "mesh" and dotted scattering were seen in fig. 8-10, and some were connected to form a band; the positive control group showed blue collagen fibers scattered in spots and partially connected into a band, and the deposition of interstitial collagen fibers in myocardium was also reduced (see FIGS. 8-11 and 8-12).

11. Results of the experiment

(1) General conditions: 1 death patient dies on the 2 nd day after the administration of the model control group, 2 abnormal heart rhythms are found by continuous 5min electrocardio detection at 2 weeks after the administration, 1 death patient dies on the 9 th day after the administration of the positive control group, and 1 death patient dies on the 9 th day after the administration of the positive control group; the low, medium and high dose groups given sulcardine sulfate did not die, nor was any abnormal heart rhythm observed.

(2) Cardiac structure: compared with a sham operation group, the IVS thickness and the LVPW thickness of the model control group in the systolic period and the diastolic period at 1 week and 2 weeks after the administration are obviously increased (P <0.01 and P < 0.05); compared with the model control group, the low, medium and high dose groups and the positive control group have no obvious influence on systolic phase IVS and diastolic phase IVS at 1 week after administration, and have different degrees of reduction at 2 weeks after administration, wherein the high and medium dose groups have obvious reduction of systolic phase IVS (P <0.05), and the positive control group has obvious reduction of diastolic phase IVS (P < 0.01). The high and medium dose groups showed a significant decrease in systolic LVPW at 2 weeks of dosing (P <0.05, P < 0.01).

(3) Electrocardiogram indexes: compared with the sham operation group, the QT interval time of the model control group is remarkably prolonged (P <0.05, P <0.01), and the ST segment elevation value (STE) is remarkably pressed down (P < 0.01); compared with a model control group, the low, medium and high dose groups have obvious improvement on the STE depression degree (P <0.05, P <0.01), and the medium and high dose groups have obvious improvement on the QT interval time prolongation (P <0.05, P < 0.01).

(4) Heart mass and coefficient: compared with a false operation group, the heart coefficient of the model control group, the heart coefficient of the ultrasonic measurement and calculation and the heart coefficient of the anatomical weighing are both obviously increased (P is less than 0.01); compared with a model control group, after administration for 2 weeks, the anatomical weighing method by an ultrasonic measurement and calculation method shows that the heart coefficients of the medium and high dose groups are remarkably reduced (P <0.05, P <0.01), the heart coefficients of the positive control group have a reduction trend, but the difference is not remarkable (P > 0.05).

(5) Biochemical indexes are as follows: compared with a sham operation group, the plasma NT-proBNP content is obviously increased (P <0.01) when the model control group is administrated for 2 weeks, compared with the model control group, the plasma NT-proBNP content of the medium-high dose group and the positive control group is obviously reduced (P <0.05, P <0.01), and the plasma NT-proBNP content of the low dose group is also reduced, but the difference is not obvious (P > 0.05).

(6) Observation of myocardial histopathology: HE staining results show that cells in the model control group are loosely arranged, and visible cells are broken, the myocardial fibers are thickened, interstitial edema exists, inflammatory cells infiltrate and bleeding exists; compared with a model control group, the shapes of the myocardial cells of the high, medium and low dose groups are improved to different degrees, the structure is relatively complete, and the arrangement is relatively regular; the positive control group also has improved myocardial cell morphology. Masson staining results show that the deposition of interstitial collagen fibers of the myocardium of the model control group is obviously increased, so that the myocardial cells in the myocardial tissues of rats are loose and are divided by the obviously thickened collagen fibers to be in a mesh shape, a large amount of blue collagen fibers are seriously distributed in a sheet shape, and the interstitial collagen fibers of the myocardium are deposited in a large amount; compared with a model control group, the deposition of the myocardial interstitial collagen fibers of the low, medium and high dose groups and the positive control group is reduced to different degrees, and the improvement of the medium and high dose groups is obvious; the positive control group also had a reduction in myocardial interstitial collagen fiber deposition. As a result of the collagen fiber deposition score, the total score of 5 rats was 3 points or more in 7 rats in the model control group, 4 rats, 2 rats and 3 rats were 3 points or more in 8 rats in the low, medium and high dose groups, and 2 rats were 3 points or more in 7 rats in the positive control group.

12. Discussion of the related Art

HF is a clinical syndrome of cardiac insufficiency caused by various cardiac diseases and is also the ultimate destination of most cardiovascular diseases. HF and arrhythmias are often associated with clinical use. Most antiarrhythmic drugs have the characteristics of inhibiting heart conduction, inhibiting myocardial contraction and the like, so that the clinical treatment of the drugs for treating heart failure combined arrhythmia is difficult^[1]. Therefore, the test observes the influence of the antiarrhythmic sulcardine sulfate on the HF rat and provides reference for clinical application.

According to the occurrence and development process of heart failure, structural heart disease develops from the risk factors of heart failure, the symptoms of heart failure appear, and the heart failure reaches the refractory terminal stage heart failure, and can be divided into 4 stages, namely, pre-heart failure (A), pre-clinical heart failure (B), clinical heart failure (C) and refractory terminal stage heart failure (D). The definition of the heart failure in the stage A is that the patient is a high-risk group of the heart failure, does not have abnormal heart structure or function, and does not have symptoms and/or physical signs of the heart failure; phase B Heart failure is defined as the patient's progression from symptoms and/or signs of no heart failure, but has progressed to a structural heart disease^[2]. The rat myocardial hypertrophy model caused by aortic arch constriction adopted in the test is an ideal model for the pathophysiological development process from myocardial compensatory hypertrophy to uncompensated heart failure caused by reduction pressure overload, the pathological change degree is controllable, the model is stable and good in repeatability, and the induced model presents a progressive heart failure process. The laboratory earlier stage research shows that the cardiac muscle hypertrophy appears 4 weeks after rat TAC operation, which is characterized by left ventricular wall thickening, ventricular cavity reduction, impaired cardiac contraction and relaxation function and compensatory hypertrophy of cardiac muscle. At 8 weeks, the myocardium may develop an initial decompensated response, with no further thickening of the ventricular walls and further impairment of cardiac function. At 12 weeks, severe fibrosis of the myocardium and thinning of the ventricular wall lead to refractory end-stage heart failure^[3]. Thus, the starting point for administration in this trial was 4 weeks post-TAC with ejection fraction retention, but ejectionThe pathological period of ventricular remodeling corresponds to clinical stage B heart failure, and belongs to an early heart failure animal model.

Echocardiogram is a common clinical noninvasive detection method, and M-type ultrasonic detection can be used for dynamically observing myocardial thickening and cardiac function change, and IVS and LVPW reflect ventricular remodeling. The experiment shows that after TAC operation for 4 weeks, IVS and LVPW of a model control group rat are obviously thickened, the rat is prompted to have left ventricle reconstruction, after 2 weeks of administration of high-dose sulcardine sulfate, the thickness of IVS and LVPW is reduced, and the sulcardine sulfate is prompted to have a certain effect of improving ventricle reconstruction.

The electrocardiogram is also a common clinical noninvasive detection method and can diagnose various pathological states such as arrhythmia, myocardial ischemia and the like. Clinical studies show that the first onset of acute left heart failure is ST-segment depression, which is caused by myocardial ischemia for a long time and has wide damaged area, so that the S-T segment depression common in electrocardiogram can be seen, and myocardial infarction, necrotic myocardial interstitial congestion, edema and reduced compliance can be further caused, and diastolic heart failure is caused^[4]. In addition, the electrophysiological remodeling of cardiac muscle during heart failure is the basis for the occurrence of arrhythmia, and is most prominently characterized by slow repolarization of cardiac muscle, prolonged action potential, and prolonged QT interval on the body surface electrocardiogram, and is regarded as acquired long QT syndrome (LQTS)^[5]. In the experiment, the electrocardio-interval of the rats in the model control group is prolonged, the ST segment is depressed, the QT interval is prolonged, and the rats after TAC operation are prompted to have myocardial infarction to a certain extent and have myocardial electrophysiological reconstruction. After the sulcardine sulfate is administered without dose, the sulcardine sulfate has obvious improvement effect on ST-segment depression and QT interval prolongation abnormity, and the sulcardine sulfate is suggested to regulate heart rhythm and resist myocardial infarction.

NT-proBNP detection is often used in combination with echocardiography, to infer the likelihood of having HF.

NT-proBNP is derived from the cardiac muscle cells, is present in the secretory granules of the cardiac muscle cells in the form of pro-BNP, and is cleaved into NT-proBNP and BNP when the pressure of the ventricular wall changes. Therefore, NT-proBNP is closely related to the systolic-diastolic function of the heart^[6,7]. The research of the experiment finds thatThe plasma NT-proBNP level of the model control group rat is obviously increased, and the rats have descending trends of different degrees after being administered with different doses of sulcardine sulfate, which indicates that the sulcardine sulfate has certain improvement effect on the heart failure state of the rats after TAC operation.

The deposition of interstitial collagen in cardiac muscle cells and myocardial fibrosis are the main factors causing the decrease of myocardial compliance, when the long-term pressure load of heart is too heavy, the content of collagen in heart and the density of collagen network are also excessively increased along with the occurrence and development of myocardial cell hypertrophy, the crosslinking degree of the myocardial collagen is proportional to the myocardial stiffness and the diastolic function, and the collagen I with stronger tensile capacity is taken as the main factor^[6]. Mason staining enables the collagen to appear blue, and is used for detecting the collagen deposition condition in the tissues. The gross anatomy and pathological diagnosis show that the rats in the model control group have myocardial hypertrophy, increased heart weight, myocardial cell edema, enlarged cell nucleus, disordered arrangement, loss of basic structure of blood vessels, hyperplasia of connective tissue of outer membrane, infiltration and centripetal diffusion of inflammatory cells and obvious collagen deposition, and after different doses of sulcardine sulfate are given, the heart weight and the heart coefficient of the rats are reduced to different degrees, and the collagen deposition state is slightly improved.

In conclusion, the sulcardine sulfate has certain improvement effect on myocardial collagen deposition and ventricular remodeling of a rat model with early heart failure.

13. Conclusion

The experimental results show that the sulcardine sulfate has no obvious influence on the heart function of the rat model with early heart failure, and has certain improvement effect on myocardial collagen deposition and ventricular remodeling of the rat model with early heart failure.

14. Reference to the literature

[1] Chenfei, liu fan, medication for heart failure combined with arrhythmia [ J ] clinical meta-extract, 2017,32 (03): 201-204.

[2] Royal ji [ china guidelines for heart failure diagnosis and treatment 2014 ] interpretation [ J ] chinese journal of clinicians 2016,44 (05): 14-16.

[3] Xi xiao qing, chen xiao zhen, majoxin, zhangli palm, fangming bamboo shoot, chenchengcheng, cai yueqin, chen foley. 35-40+85.

[4] Lie. general ST-segment depressed myocardial infarction with acute left heart failure as the first manifestation 20 clinical analyses [ J ]. journal of practical cardio-pulmonary vascular disease, 2009,17 (9): 794-795.

[5]Choy A M,Lang C C,Chomsky D M,et al.Normalization of acquired QT prolongation in humans by intravenous potassium[J].Circulation,1997,96(7)：2149-2154.)

[6] Study on relationship between cardiac hypertrophy index and cardiac function of heart failure model rats [ J ] proceedings of first university of medicine, 2010 (5): 596-599.

The invention provides a thinking and a method for applying sulcardine sulfate in preparing anti-heart failure products, and a method and a way for realizing the technical scheme are many, the above description is only a preferred embodiment of the invention, and it should be noted that, for a person skilled in the art, a plurality of improvements and decorations can be made without departing from the principle of the invention, and the improvements and decorations should also be regarded as the protection scope of the invention. All the components not specified in the present embodiment can be realized by the prior art.