Application of Xinsunning preparation in preparation of medicine for preventing or treating vagus nerve atrial fibrillation

Abstract

The invention belongs to the technical field of medicine application, and relates to an application of a Xinsunning preparation in preparing a medicine for preventing or treating vagus nerve atrial fibrillation, the Xinsunning preparation reduces the occurrence rate of vagus nerve atrial fibrillation, shortens the time of vagus nerve atrial fibrillation, reduces the P-wave time history of vagus nerve atrial fibrillation, and improves the treatment effect of the vagus nerve atrial fibrillation. The sympathetic nerve tension and the sympathetic nerve activity are obviously reduced, and the vagus nerve tension and the vagus nerve activity are obviously improved, so that the vagus nerve atrial fibrillation is prevented or treated, and the dosage of the Xinxin preparation for preventing or treating the vagus nerve atrial fibrillation is 0.041 g / kg.d to 0.165 g / kg.d. The invention provides a new way for the application of the Xinsunning preparation in preparation of medicines for preventing and treating vagus nerve atrial fibrillation.

Description

Technical Field

[0001] The invention belongs to the technical field of drug application, and relates to the use of a xinsuning preparation in preparing a drug for preventing or treating vagal atrial fibrillation. Background Art

[0002] Xinsu Ning Capsule (National Medicine Standard No. Z20050131, Shaanxi Momed Qixuehe Pharmaceutical Co., Ltd.) is composed of 11 Chinese medicinal herbs: Coptis chinensis, Pinellia ternata, Poria cocos, Citrus aurantium, Dichroa chinensis, Lotus seed core, Sophora flavescens, Cyathula capitata, Ginseng, Ophiopogon japonicus, and Licorice. It is an effective prescription for the treatment of phlegm-heat disturbing heart type arrhythmia (premature beats) in clinical practice, and has the effects of clearing away heat and resolving phlegm, calming the heart and relieving palpitations. It is used for palpitations, chest tightness, irritability, easy startle, dry mouth and bitter taste, insomnia, dizziness, and irregular pulse caused by phlegm-heat disturbing the heart; mild and moderate ventricular premature beats caused by coronary heart disease and viral myocarditis with the above symptoms.

[0003] Atrial fibrillation (AF) is a supraventricular tachyarrhythmia with uncoordinated atrial electrical excitation and ineffective atrial contraction. Clinically, the types of AF are very complex, and multiple factors are involved in its formation. "Atrial Fibrillation: Current Understanding and Treatment Recommendations (2021)" lists two of the most recognized mechanisms, namely electrophysiological mechanisms and pathophysiological mechanisms, which include atrial remodeling, neural regulation, and genetic influences. The causes of AF are different, and the response to drug treatment is also different. For example, dofetilide is very effective in terminating persistent AF in dogs associated with chronic heart failure, but it is very ineffective for tachycardia-induced remodeling AF; propafenone is effective for newly occurring AF, but has poor efficacy for persistent AF and atrial flutter; dronedarone is only effective after paroxysmal and persistent AF is converted to sinus rhythm, but it cannot be used for permanent AF, heart failure, and AF caused by left ventricular systolic dysfunction. Therefore, therapeutic drugs that work on one type of AF cannot be fully extended to other types of AF in clinical practice.

[0004] In 1978, Coumel proposed the concept of vagal atrial fibrillation. Vagal atrial fibrillation (Vagal AF) is a type of atrial fibrillation, which usually occurs together with paroxysmal atrial fibrillation and is usually considered a benign condition, but it may also develop into persistent atrial fibrillation. The characteristics of vagal atrial fibrillation are: 1. The attacks often occur at night or during rest, and rarely or never occur during physical activities or emotional excitement. 2. Atrial fibrillation attacks are often related to eating, especially after dinner. Because after dinner, the vagus nerve is not only in the excitement and stimulation of the digestive period, but also in the transition from day to night, when the vagus nerve tension gradually increases; the alcohol absorption period after drinking is also a triggering factor. The attacks of most patients are related to the gastric digestion period after meals, and a few patients' atrial fibrillation is directly related to swallowing during eating and food irritating the pharynx and esophagus. 3. The attacks often end in the early morning or early morning, when the excitability of the vagus nerve decreases and the excitability of the sympathetic nerve is relatively enhanced.

[0005] At present, the main methods for treating vagal atrial fibrillation include drugs, catheter ablation and surgery. Ion channel blockers under electrophysiological mechanism research cannot cover the treatment range of vagal atrial fibrillation. Most antiarrhythmic drugs are ineffective, which is a characteristic of vagal atrial fibrillation. In Western medicine, the use of digitalis or digoxin for treatment promotes its occurrence. The use of β-blockers can shorten the duration of each vagal atrial fibrillation, but at the same time, it can make the attack more frequent. At the same time, there is no relevant report on the treatment of vagal atrial fibrillation with Chinese patent medicine. Since its launch, Xinsuning preparations have been proven safe and effective for rapid arrhythmias due to their multi-component, multi-target, multi-pathway and systemic regulation characteristics, and have outstanding advantages. Xinsuning capsules mainly exert anti-arrhythmic effects through the electrophysiological mechanism of blocking multiple ion channels (NaV1.5, CaV1.2, hERG, etc.) and regulating non-ionic targets (NNI3, CACNA1S, SCN5A, etc.). Xinsuning preparations have great application value and prospects in the clinical treatment of vagal atrial fibrillation. Summary of the invention

[0006] In view of this, the present invention aims to solve one of the technical problems in the related art at least to a certain extent. The present invention provides a use of a Xinsuning preparation in preparing a drug for preventing or treating vagal atrial fibrillation.

[0007] The present invention prevents or treats vagal atrial fibrillation by utilizing the xinsuning preparation to reduce the incidence of vagal atrial fibrillation, shorten the duration of vagal atrial fibrillation and reduce the duration of P waves.

[0008] In order to achieve the above object, the technical solution adopted by the present invention is:

[0009] A use of a xinsuning preparation in preparing a drug for preventing or treating vagal atrial fibrillation.

[0010] The Xinsuning preparation is used in preparing drugs for preventing or treating vagal atrial fibrillation by reducing the incidence of vagal atrial fibrillation.

[0011] The invention discloses a use of the Xinsuning preparation in preparing a medicine for preventing or treating vagal atrial fibrillation by shortening the time of vagal atrial fibrillation.

[0012] The invention discloses a use of the Xinsuning preparation in preparing a drug for preventing or treating vagal atrial fibrillation by reducing the P wave duration.

[0013] The invention discloses a use of the Xinsuning preparation in preparing a medicine for preventing or treating vagal atrial fibrillation by reducing sympathetic nerve tension and sympathetic nerve activity.

[0014] The invention discloses a use of the Xinsuning preparation in preparing a medicine for preventing or treating vagal atrial fibrillation by increasing vagal nerve tension and vagal nerve activity.

[0015] It is understandable that in some patients with vagal atrial fibrillation, vagal nerve stimulation may shorten the wavelength by shortening the atrial effective refractory period (ERP), thereby creating conditions for the occurrence of reentry. The specific effect of acetylcholine ACh on specific cholinergic synapses depends to a certain extent on the type of ACh receptors on the synapses. The main distribution in the atrial muscle is the M2 subtype of muscarinic cholinergic receptors (mAChR). ACh secreted by the parasympathetic nervous system acts on the M2 receptors in the atrial muscle and then activates the β and γ subunits of G proteins, thereby interacting with ACh-dependent K+ current (I KACh ) combined, resulting in I KACh Open, so I KACh Also known as G protein-gated inwardly rectifying K (GIRK) channels, the activation of GIRK leads to K + The outflow of vagus nerve fibers in the atrial cavity shortens the duration of the action potential (AP) and the ERP. In addition, since the distribution of atrial vagal nerve fibers is uneven, the increased excitability of the vagus nerve causes an increase in the dispersion of the atrial ERP, which can enhance the sensitivity of the atrium to reentrant movement, thereby inducing vagal atrial fibrillation.

[0016] Xinsuning preparations can improve the regulatory mechanism of vagal atrial fibrillation. They can effectively improve the electrocardiogram manifestations of rats with vagal atrial fibrillation, reduce the incidence of vagal atrial fibrillation in rats, shorten the duration of vagal atrial fibrillation, and reduce the duration of P waves. They can significantly reduce the sympathetic nerve tone and sympathetic nerve activity, and significantly increase the vagal nerve tone and vagal nerve activity, thereby preventing or treating vagal atrial fibrillation.

[0017] It is further specified that the dosage of the Xinsuning preparation for preventing or treating vagal atrial fibrillation is 0.041 g / kg.d to 0.165 g / kg.d.

[0018] It is further specified that the Xinsuning preparation is prepared by combining the following raw materials in proportion by mass: the mass ratio of Coptis chinensis, Pinellia ternata, Poria cocos, Immature Citrus aurantium, Dichroa febrifuga, Lotus seed core, Sophora flavescens, Artemisia annua, Ginseng, Ophiopogon japonicus and Licorice is 1:0.745:0.745:0.496:0.745:0.135:0.745:0.745:0.496:0.745:0.496.

[0019] It is further defined that the Xinsuning preparation is prepared by the following steps:

[0020] (1) Purify, slice, and dry the raw materials of coptis root, pinellia tuber, tuckahoe, immature bitter orange, radix chebulae, lotus seed core, sophora flavescens, artemisia annua, ginseng, ophiopogon japonicus, and liquorice according to the mass ratio;

[0021] (2) Take coptis root, immature bitter orange, radix chebulae, lotus seed core and sophora flavescens, add 60% ethanol, reflux extraction twice, each time for 1.5 hours, combine the extracts, filter, reduce the pressure of the filtrate to recover ethanol, continue to concentrate to a thick paste with a relative density of 1.36 to 1.38 at 60°C, dry at 80°C to form a dry paste, and discard the residue;

[0022] (3) Take ginseng, Poria cocos, and Pinellia tuber and add 70% ethanol, reflux extract twice, each time for 2 hours, combine the extracts, filter, and collect the filtrate in another container for later use; set aside the residue for later use;

[0023] (4) The residue obtained in step (3) is combined with Ophiopogon japonicus, Artemisia annua and Glycyrrhiza uralensis, and decocted twice with water, each time for 1 hour, the decoctions are combined, filtered, and the filtrate is concentrated to a clear paste with a relative density of 1.05 to 1.06 at 80° C., ethanol is added to make the alcohol content 70%, stirred evenly, and allowed to stand for 24 hours; the supernatant is taken, filtered, and the filtrate is combined with the filtrate collected in another device in step (3), the ethanol is recovered under reduced pressure, and the mixture is further concentrated to a thick paste with a relative density of 1.36 to 1.38 at 60° C., and dried at 80° C. to obtain a dry paste;

[0024] (5) The two dry pastes obtained in step (2) and step (4) are combined, crushed into fine powder, and other pharmaceutical excipients are added, mixed, and prepared into a preparation.

[0025] It is further defined that the Xinsuning preparation of the present invention is prepared according to conventional pharmaceutical methods, including oral liquid preparations such as tablets, capsules, granules, pills, etc., and auxiliary materials used in pharmacy are added during the preparation process, such as: fillers, binders, disintegrants, lubricants, flavoring agents, etc.

[0026] Compared with the prior art, the present invention has the following beneficial effects:

[0027] 1. Compared with the existing Xinsuning capsule which mainly blocks multiple ion channels and regulates non-ionic targets, the present invention has found that the Xinsuning preparation can effectively improve the electrocardiogram performance of rats with vagal atrial fibrillation, reduce the incidence of vagal atrial fibrillation in rats, shorten the duration of vagal atrial fibrillation and reduce the P wave duration of vagal atrial fibrillation, significantly reduce sympathetic nerve tone and sympathetic nerve activity, and significantly increase vagal nerve tone and vagal nerve activity, thereby preventing or treating vagal atrial fibrillation.

[0028] 2. In the present invention, with the increase of the dosage of the Xinsuning preparation, the incidence of vagal atrial fibrillation and the duration of atrial fibrillation are significantly reduced. When the dosage of the Xinsuning preparation is 1.04g / kg.d (high dose), the incidence of vagal atrial fibrillation is 0, and the mean duration of atrial fibrillation is 0s; while the P wave duration increases with the increase of the dosage; when it is 0.26g / kg.d (low dose), the mean duration of the P wave is 0.0169s, which is close to the blank group of 0.016s and is 0.006s lower than that of the model group, indicating that the dosage adjustment of the Xinsuning preparation has a good preventive and therapeutic effect on vagal atrial fibrillation.

[0029] 3. The present invention uses an ACh-CaCl2 model to simulate the effect of vagal overexcitation on the heart. Compared with other atrial fibrillation models, the Ach in the model used in the present invention is acetylcholine, which is a neurotransmitter of the vagus nerve and acts on the M2 receptors on the heart; high concentrations of Ach simulate excessive activation of the vagus nerve, which can better correspond to vagal atrial fibrillation, making the research process of the present invention more scientific and accurate.

[0030] 4. The present invention verifies the mechanism of action of xinsuning in treating vagal atrial fibrillation by adopting the ACh-CaCl2 model, fills the technical gap of xinsuning preparations in improving vagal atrial fibrillation, finds a new approach for the research on the treatment of vagal atrial fibrillation, realizes a new idea for the new use of old drugs, saves the new drug development process, reduces costs, and ensures safe and reliable medication. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 Flow chart for modeling vagal atrial fibrillation in rats;

[0032] Figure 2 The electrocardiograms of rats in each group are shown in Figure 2.

[0033] Figure 3 This is a comparison chart of the incidence of vagal atrial fibrillation in rats;

[0034] Figure 4 This is a comparison chart of the duration of vagal atrial fibrillation in rats;

[0035] Figure 5 This is a comparison of the P wave duration of the rat electrocardiogram;

[0036] Figure 6 This is a comparison chart of heart rate variability in rats;

[0037] Figure 7 This is a recording of vagus nerve activity in rats;

[0038] Figure 8 This is a recording of sympathetic nerve activity in rats;

[0039] Fig. 9 This is a comparison chart of the vagus nerve and sympathetic nerve activities in rats. DETAILED DESCRIPTION

[0040] The present invention will now be described in detail with reference to the accompanying drawings and specific implementation methods.

[0041] The invention develops a use of a Xinsuning preparation in preparing a drug for preventing or treating vagal atrial fibrillation. The Xinsuning preparation is prepared by the following raw materials in mass proportion: the mass ratio of coptis root, pinellia tuber, tuckahoe, immature bitter orange, radix changshan, lotus seed heart, sophora flavescens, artemisia annua, ginseng, ophiopogon japonicus and liquorice is 1:0.745:0.745:0.496:0.745:0.135:0.745:0.745:0.496:0.745:0.496.

[0042] Xinsuning preparations can prevent or treat vagal atrial fibrillation by reducing the incidence of vagal atrial fibrillation, shortening the duration of vagal atrial fibrillation, and reducing the duration of P waves, significantly reducing sympathetic nerve tone and sympathetic nerve activity, and significantly increasing vagal nerve tone and vagal nerve activity.

[0043] The dosage of Xinsuning preparation for preventing or treating vagal atrial fibrillation is 0.041g / kg.d to 0.165g / kg.d.

[0044] The present invention is described below with reference to specific embodiments. It should be noted that these embodiments are merely illustrative and do not limit the present invention in any way.

[0045] Example 1 Preparation of Xinsuning Capsules

[0046] Composition formula: 334g of Coptis chinensis, 250g of Pinellia ternata, 250g of Poria cocos, 167g of Immature Citrus aurantium, 250g of Dichroa chinensis, 42g of Lotus seed core, 250g of Sophora flavescens, 250g of Artemisia annua, 167g of Ginseng, 250g of Ophiopogon japonicus, and 167g of Licorice.

[0047] The process steps are as follows:

[0048] (1) The raw materials are cleaned, sliced, and dried for later use.

[0049] (2) 334 g of Coptis chinensis, 167 g of Citrus aurantium, 250 g of Radix Dichroae, 42 g of Pellets of Nelumbo nucifera and 250 g of Sophora flavescens were added with 60% ethanol and refluxed for 2 times, each time for 1.5 hours. The extracts were combined and filtered, and the filtrate was decompressed to recover ethanol, and the filtrate was further concentrated to a thick paste with a relative density of 1.36 to 1.38 at 60° C., and dried at 80° C. to obtain a dry paste, and the residue was discarded;

[0050] (3) Take 167g of ginseng, 250g of Poria cocos and 250g of Pinellia ternata, add 70% ethanol, reflux extract twice, each time for 2 hours, combine the extracts, filter, and collect the filtrate in another container for later use; the residue is set aside for later use;

[0051] (4) The residue obtained in step (3) is combined with 250 g of Radix Ophiopogonis, 250 g of Artemisia annua and 167 g of Radix Glycyrrhizae, and decocted twice with water, each time for 1 hour. The decoctions are combined, filtered, and the filtrate is concentrated to a clear paste with a relative density of 1.05 to 1.06 at 80°C. Ethanol is added to make the alcohol content 70%, stirred evenly, and allowed to stand for 24 hours. The supernatant is taken, filtered, and the filtrate is combined with the filtrate collected in another device in step (3), and the ethanol is recovered under reduced pressure. The mixture is further concentrated to a thick paste with a relative density of 1.36 to 1.38 at 60°C, and dried at 80°C to obtain a dry paste.

[0052] (5) Combine the two dry pastes obtained in step (2) and step (4), crush them into fine powder, add appropriate amount of dextrin, mix well, put them into capsules, and make 1000 capsules.

[0053] Example 2 Preparation of Xinsu Ning Tablets

[0054] The composition formula of Xinsu Ning Tablets is the same as that of Xinsu Ning Capsules, and the process steps (1) to (4) are also the same as those of Xinsu Ning Capsules, except that:

[0055] (5) The two dry pastes obtained in step (2) and step (4) are combined, crushed, added with an appropriate amount of dextrin, granulated with 85% ethanol, dried at 60°C, granulated, added with 0.5% magnesium stearate, mixed well, pressed into 1000 tablets, and coated.

[0056] Example 3 Preparation of Xinsu Ning Pills

[0057] The composition formula of Xinsu Ning Pills is the same as that of Xinsu Ning Capsules, and the process steps (1) to (4) are also the same as those of Xinsu Ning Capsules, except that:

[0058] (5) The two dry pastes obtained in step (2) and step (4) are combined and crushed, and 35-50 g of refined honey and an appropriate amount of water are added to every 100 g of powder to make pills, and the pills are dried to make water-honey pills; or 80-130 g of refined honey are added to make small honey pills or large honey pills.

[0059] Example 4 Preparation of Xinsuning Granules

[0060] The composition formula of Xinsu Ning Granules is the same as that of Xinsu Ning Capsules, and the process steps (1) to (4) are also the same as those of Xinsu Ning Capsules, except that:

[0061] (5) The two dry pastes obtained in step (2) and step (4) are combined, and an appropriate amount of sucrose powder is added to form granules, and dried to obtain the granules.

[0062] In the preparation process, auxiliary materials used in pharmacy such as fillers, binders, disintegrants, lubricants, flavoring agents and the like may also be added to oral liquid preparations.

[0063] Example 5

[0064] The Xinsu Ning capsules prepared in Example 1 were used to carry out the following experimental process.

[0065] 5.1 Experimental Materials

[0066] 5.1.1 Experimental instruments

[0067] The instruments used in this embodiment are shown in Table 1.

[0068] Table 1 Main experimental instruments

[0069]

[0070] 5.1.2 Experimental reagents

[0071] Xinsuning capsules were purchased from Shaanxi Momede Qixuehe Pharmaceutical Co., Ltd. (approval number: National Medicine Standard Z20050131).

[0072] Amiodarone, item number A8423-10G, specification 100 g, purchased from SIGMA. Acetylcholine, item number A2661, specification 100 g, purchased from SIGMA. Calcium chloride (CaCl2) C1016, specification 100 g, purchased from SIGMA.

[0073] 5.1.3 Experimental animals

[0074] Fifty male CD (SD) rats, weighing 250±20 g, were housed in the SPF standard animal breeding room of the Experimental Animal Center of Shandong University of Traditional Chinese Medicine.

[0075] Feeding conditions: temperature controlled at 24 ± 2 ° C; humidity controlled at 50 ± 10%; artificial light 12h / d; ventilation rate 15 times / h, rats free access to water and food. The rats used in the experiment were purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd., animal production license number: SCXK (Beijing) 2021-0011. All animals in the experiments were strictly in accordance with the "Guiding Principles for the Care and Use of Laboratory Animals" of Shandong University of Traditional Chinese Medicine (Animal Use License: SYXK (Beijing) 2021-0011). This study has been approved by the Animal Ethics Review Committee of Shandong University of Traditional Chinese Medicine.

[0076] 5.2 Experimental methods

[0077] 5.2.1 Grouping

[0078] The experiment was divided into the following six groups: control group (CON group), vagal atrial fibrillation model group (AF group), amiodarone treatment group (AF+AMD group), low-dose treatment group of xinsuning preparation (AF+XSN(L) group, xinsuning dose 0.26g / kg / d), medium-dose treatment group of xinsuning preparation (AF+XSN(M) group, xinsuning dose 0.52g / kg / d) and high-dose treatment group of xinsuning preparation (AF+XSN(H group, xinsuning dose 1.04g / kg / d).

[0079] Referring to the equivalent dose coefficient conversion method appendix of "Pharmacological Experimental Methodology" edited by Professor Xu Shuyun, the equivalent dose ratio table between humans and animals converted by body surface area, the human clinical dose is X mg / kg, and the dose formula converted to rats is: X mg / kg×70kg×0.018 / 200g=X mg / kg×70kg×0.018 / 0.2kg=6.3X mg / kg. The equivalent dose of rats is 6.3 times that of humans. The low-dose treatment group of the xinsuning preparation (AF+XSN(L) group, xinsuning dose of 0.26g / kg / d) was equivalent to the human dose of 0.041g / kg / d; the medium-dose treatment group of the xinsuning preparation (AF+XSN(M) group, xinsuning dose of 0.52g / kg / d) was equivalent to the human dose of 0.082g / kg / d; the high-dose treatment group of the xinsuning preparation (AF+XSN(H) group, xinsuning dose of 1.04g / kg / d) was equivalent to the human dose of 0.165g / kg / d.

[0080] 5.2.2 Drug preparation

[0081] AF+XSN(L) group: weigh an appropriate amount of the contents of Xinsuning capsules and add water to prepare a 0.26 g / kg suspension aqueous solution.

[0082] AF+XSN(M) group: weigh an appropriate amount of the contents of Xinsuning capsules and add water to prepare a 0.52 g / kg suspension aqueous solution.

[0083] AF+XSN(H) group: weigh an appropriate amount of the contents of Xinsuning capsules and add water to prepare a 1.04 g / kg suspension aqueous solution.

[0084] AF+AMD group: Weigh 50 mg of amiodarone hydrochloride and dissolve it in 10 mL of water to prepare a suspension aqueous solution with a concentration of 5.0 mg / mL.

[0085] Preparation method of calcium chloride acetylcholine (ACh-CaCl2) mixed solution: Dissolve 1.51g of CaCl2 and 10mg of ACh in 151mL of 0.9% NaCl solution (normal saline), and immediately divide into 3 EP tubes. Prepare the model drug freshly every day, keep it away from light during the preparation process, and store it on ice for later use.

[0086] 5.2.3 Atrial Fibrillation Modeling

[0087] The commonly used atrial fibrillation models are as follows: ① Aconitine atrial fibrillation model: 5 μL of 0.03% aconitine was locally injected at any location of the right auricle or left pulmonary vein of the dog, followed by 5 stimulations of 5 seconds near the injection site, which successfully induced persistent atrial fibrillation. This model can be used for the study of focal atrial fibrillation. ② Electrically stimulated rapid atrial pacing atrial fibrillation model: A higher frequency of electrical stimulation was used to repeatedly stimulate the right atrium of the goat to establish a persistent and stable chronic atrial fibrillation model. This type of atrial fibrillation is currently the most studied and is widely used in the study of the mechanism, electrophysiological and structural changes, and treatment of chronic atrial fibrillation. ③ Mitral regurgitation atrial fibrillation model: The femoral artery of the experimental dog was hooked with a catheter with a hook knife to cut the mitral valve key cable, causing moderate to severe regurgitation. After a few weeks, atrial burst stimulation was given, and 52.6% of the dogs had persistent atrial fibrillation. This model can be used for electrophysiological, pathological and therapeutic studies of valvular disease combined with atrial fibrillation. Compared with the above models, the ACh-CaCl2 model used in the present invention simulates the effects of excessive vagal nerve excitation on the heart. A rat vagal atrial fibrillation model was prepared by tail vein injection of ACh-CaCl2. On the one hand, ACh was injected intravenously to increase the ACh concentration in the rat body. This rapidly activated GIRK, resulting in shortening of AP and ERP, and vagal atrial fibrillation. On the other hand, CaCl2 was used to cause transient calcium overload in atrial myocytes, resulting in shortening of atrial action potential duration (APD) and wavelength (WL). The specific steps of this study are:

[0088] CD (SD) rats were injected with a mixture of ACh and CaCl2 (66 μg + 10 mg / mL, 1 mL kg -1 The solution was prepared immediately before use and kept away from light. The prepared modeling drug was placed in an ice box away from light. The CD (SD) rats in the control group were injected with an equal amount of saline (1 mL kg -1 The specific process is as follows: put the rat into the anesthesia induction box to induce anesthesia (concentration 5%, ventilation volume 1Lvmin -1 ) for 3 minutes, and then maintain anesthesia with a mask (concentration 2%, ventilation volume 0.4 L min -1 ) Place the rat in supine position on a heating pad and wipe it gently and repeatedly with an alcohol cotton ball. Start looking for the vein from the distal end of the rat's tail, i.e. the tip of the tail. Use a 1mL insulin syringe to extract the CaCl2 and ACh mixture. The air in the syringe must be exhausted to avoid death of the rat due to injection of air. Hold the tail with your fingers and insert the needle from the distal end at an angle of 10° to 15°. The depth of the needle varies depending on the position of the rat's tail. When the needle enters the blood vessel, there will be a sense of emptiness and the syringe needs to be withdrawn. If there is blood returning, it indicates that the needle is in the vein. Keep the needle inserted steadily before injection. The injection should be completed within 5 seconds. Repeat the above process once a day at the same time period for 10 consecutive days.

[0089] 5.2.4 Drug administration

[0090] The drug was administered by intragastric gavage at a dose of 1 mL kg -1 (Control group: normal saline; AF+XSN(L) group; AF+XSN(M) group; AF+XSN(H) group; AF+AMD group).

[0091] 5.2.5 Experimental process

[0092] See also Figure 1 The specific experimental process used in this embodiment is as follows.

[0093] Day 1: ACh-CaCl2 modeling;

[0094] Day2: ACh-CaCl2 modeling;

[0095] Day 3: ACh-CaCl2 modeling, ECG monitoring;

[0096] Day 4: ACh-CaCl2 modeling and drug gavage;

[0097] Day5: ACh-CaCl2 modeling and drug gavage;

[0098] Day6: ACh-CaCl2 modeling, drug gavage;

[0099] Day7: ACh-CaCl2 modeling and drug gavage;

[0100] Day8: ACh-CaCl2 modeling and drug gavage;

[0101] Day9: ACh-CaCl2 modeling and drug gavage;

[0102] Day 10: ACh-CaCl2 modeling, drug gavage, and electrocardiogram monitoring.

[0103] 5.2.6 Electrocardiogram test

[0104] The specific process of electrocardiogram detection is as follows: each group of rats is placed in the anesthesia induction box of the anesthesia machine to induce anesthesia (concentration 5%, ventilation volume 1L / min) for 3 minutes, and then anesthesia is maintained with a mask (concentration 2%, ventilation volume 0.4L / min). The rats are fixed on a wooden board in a supine position, and the needles are inserted subcutaneously. The acupuncture needles are inserted into the right upper limb, right lower limb, and left lower limb of the rat in a parallel and centripetal manner. The needle handles are connected to the PowerLab 8 / 35 multi-channel physiological instrument electrocardiogram electrodes, with the right upper limb as the negative electrode, the right lower limb as the ground wire, and the left lower limb as the positive electrode. Record the standard limb II lead electrocardiogram. After the electrocardiogram is connected, observe the static electrocardiogram for 10 minutes. If an abnormal heart rhythm occurs, it will be eliminated. The appearance of f waves and the disappearance of p waves are used as signs of AF occurrence, and the disappearance of f waves and the appearance of p waves are used as signs of AF termination. The induction time of vagal atrial fibrillation and the duration of vagal atrial fibrillation are recorded.

[0105] 5.2.7 Analysis of heart rate variability (HRV) in rats

[0106] Sympathetic and parasympathetic control of the cardiovascular system determines the oscillatory responses of heart rate and blood pressure to exogenous and endogenous stimuli. These rhythmic components can be analyzed and quantified using heart rate variability (HRV). Heart rate variability (HRV) refers to the variation in the difference between the heart beats and cycles. To investigate the effect of electroacupuncture of the Xinsuning preparation on heart rate variability in rats with vagal atrial fibrillation, the rats in each group were fixed on an animal heating pad in a standard supine position, and the limbs were fixed with tape. The electrocardiogram (5 mm / mV, 1 k / s sampling rate) including atrial fibrillation p-waves, RR intervals, and f-waves was recorded using a multi-channel physiological recorder (AD Instrument PowerLab 8 / 35, Australia) for 5 minutes. The frequency domain power spectral density analysis method is a commonly used HRV analysis method. Spectral analysis provides information on the frequency and amplitude of specific rhythms in the HRV waveform, and these oscillations can be quantified in any given cycle. The ratio of low frequency to high frequency (LF / HF) is usually used to evaluate the balance of sympathetic and vagal nerves: LF is 0.2-0.75Hz, and HF is 0.75-2.5Hz. The ECG data were recorded and analyzed using the LabChart 8.0 system (ADInstrument PowerLab 8 / 35, Australia) with a sampling frequency of 4k / s. The incidence of vagal AF, AF duration, standard deviation of neural network interval (SDNN), and P-wave duration were calculated. The typical ECG manifestation of vagal AF is arrhythmia, with the presence of F waves and the disappearance of P waves.

[0107] 5.2.8 Multi-fiber recording of sympathetic and vagus nerve activity in the neck

[0108] Sympathetic and vagus nerve activity in the neck was directly measured by multi-fiber nerve recording. Rats were anesthetized with 2% isoflurane and placed in the supine position on a heating pad with body temperature maintained at (37 ± 2) °C. The hair of the neck was cleaned and the skin and muscle were separated by a midline incision. The trachea and left carotid artery were separated under a microscope using a glass separation needle. The sympathetic nerves are located on the medial side of the carotid artery, while the vagus nerve is located on the lateral side. The sympathetic and vagus nerves were fixed with a glass separation needle and the epineurium was carefully peeled off with fine forceps. A small amount of liquid paraffin was added to keep the nerve moist and maintain a suitable local temperature. Then a bipolar platinum wire electrode was gently hooked around the sympathetic nerve, ensuring that the inner surface of the electrode was close to the sympathetic nerve, and the recording was performed for 20 seconds. The vagus nerve was recorded in the same way for 20 seconds. The low-pass (LP) filter was set to 100 Hz and the high-pass (HP) filter was set to 1 Hz. The GND grounding needle electrode was inserted into the deep layer of the rat's lower limb muscle, and the reference needle electrode was inserted into the subcutaneous tissue of the electrode locality; the signal was collected by the Apollo in vivo electrophysiological recording system; the in vivo electrophysiological analysis system (version: NeuroExplorer 5) was used to analyze the discharge frequency of the two autonomic nerves.

[0109] 5.2.9 Data processing

[0110] For the statistical analysis of the electrocardiogram, the noise of the preliminary record was filtered out by high-pass and low-pass filtering functions. The 5-min electrocardiogram recording segment was first selected using the Labchart8.0 analysis software, and the time of atrial fibrillation was analyzed in the Powerlab pro cardiovascular module. The incidence of atrial fibrillation in each group of SD male rats was obtained based on the number of occurrences. The P-wave was obtained from the software analysis in the selected segment, and the SDNN index was further obtained based on the RR interval.

[0111] The frequency domain data of the selected segments were analyzed from the HRV module to obtain low frequency (LF), very low frequency (VLF), and high frequency (HF). The raw data were exported and then exported into Excel format. The data were integrated and graphed using GraphPad Prism v8.0 software.

[0112] The experimental data were analyzed using SPSS26.0 software, and the statistical graphs were drawn using GraphPad Prism 8.30 software. The measured data were expressed as mean ± standard deviation (X ± SEM). One-way ANOVA was used to compare repeated measurement data of multiple groups, and LSD-t test was used to compare data between two groups. P < 0.05 was considered statistically significant. All statistical results were tested with a test statistic α = 0.05 as the significance test standard. *P < 0.05, **P < 0.01, ***P < 0.001. LF is 0.2-0.75Hz, and HF is 0.75-2.5Hz.

[0113] 5.3 Conclusion

[0114] 5.3.1 Xinsuning preparations can reduce the incidence of vagal atrial fibrillation, shorten the duration of vagal atrial fibrillation, and reduce the duration of the P wave of atrial fibrillation.

[0115] like Figure 2 The electrocardiogram results of the rats on the 10th day shown in the figure show that the control group (CON group) had obvious P waves; the P waves disappeared in the vagal atrial fibrillation model group (AF group), and continuous irregular jagged f waves appeared; the amiodarone treatment group (AF+AMD group) had obvious P waves and short-term atrial block; the low-dose treatment group of XSN preparation (AF+XSN(L) group), the medium-dose treatment group of XSN preparation (AF+XSN(M) group) and the high-dose treatment group of XSN preparation (AF+XSN(H) group) had obvious P waves.

[0116] like Figure 3The statistical results of the incidence of vagal atrial fibrillation in rats shown in the figure show that the incidence of vagal atrial fibrillation in the CON group is 0%; the incidence of vagal atrial fibrillation in the AF group is 82.61%; the incidence of vagal atrial fibrillation in the AF+AMD group is 25%; the incidence of vagal atrial fibrillation in the AF+XSN(L) group is 14.29%; the incidence of vagal atrial fibrillation in the AF+XSN(M) group is 4.76%; the incidence of vagal atrial fibrillation in the AF+XSN(H) group is 0%. Compared with the AF group, the low, medium and high dose treatment groups of Xinsuning preparation can significantly reduce the incidence of vagal atrial fibrillation in rats.

[0117] like Figure 4 The statistical results of the duration of vagal atrial fibrillation in rats shown in the figure show that the mean duration of vagal atrial fibrillation in the CON group is 0s; the mean duration of vagal atrial fibrillation in the AF group is 1.813s; compared with the AF group, the duration of vagal atrial fibrillation in the AF group has a significant statistical difference (P<0.001); the mean duration of vagal atrial fibrillation in the AF+AMD group is 0.355s; the mean duration of vagal atrial fibrillation in the AF+XSN(L) group is 0.147s; the mean duration of vagal atrial fibrillation in the AF+XSN(M) group is 0.058s, and the mean duration of vagal atrial fibrillation in the AF+XSN(H) group is 0s. Compared with the AF group, the duration of vagal atrial fibrillation in the AF+AMD group, the low, medium and high dose treatment groups of Xinsuning preparation were significantly reduced, with statistical differences (P<0.001).

[0118] like Figure 5 The statistical results of the P wave duration of rats shown in the figure show that the mean P wave duration of the CON group was 0.016s; the mean P wave duration of the AF group was 0.022s; compared with the AF group, the P wave duration of the AF group was significantly increased in the CON group, with a statistically significant difference (P < 0.01); the mean P wave duration of the AF+AMD group was 0.0189s; compared with the AF group, the P wave duration of the AF+AMD group was decreased, but there was no statistical difference (P > 0.05); the mean P wave duration of the AF+XSN (L) group was 0.0169s; the mean P wave duration of the AF+XSN (M) group was 0.0171s, and the mean P wave duration of the AF+XSN (H) group was 0.01758s. The P wave duration in the AF+XSN(L) group was significantly reduced, with statistical significance (P<0.01). The P wave duration in both the AF+XSN(M) group and the AF+XSN(H) group was reduced, with statistical significance (P<0.05).

[0119] It can be seen from this embodiment that the use of the Xinsuning preparation can effectively improve the electrocardiogram performance of rats with vagal atrial fibrillation, reduce the incidence of vagal atrial fibrillation in rats, shorten the duration of vagal atrial fibrillation, and reduce the P wave duration of vagal atrial fibrillation, indicating that the Xinsuning preparation can prevent and treat vagal atrial fibrillation.

[0120] 5.3.2 Xinsuning preparations can significantly reduce sympathetic nerve tone and significantly increase vagus nerve tone.

[0121] Heart rate variability (HRV) is a widely used measurement to assess the cardiac autonomic nervous system. Changes in the defined high frequency range (HF 0.75 Hz-2.5 Hz) reflect vagal tone, and LF 0.2 Hz-0.75 Hz reflect sympathetic tone.

[0122] like Figure 6 As shown, HRV was determined by analyzing the changes in the time / frequency domain components of VLF and HF obtained by analyzing the electrocardiogram. It was found that the LF in the AF group rats was significantly higher than that in the control group rats, with a statistical difference (P < 0.001); the AF+XSN(M) group could significantly reduce the LF, with a statistical difference (P < 0.001). The HF in the AF group rats was significantly lower than that in the control group rats, with a statistical difference. (P < 0.001); the AF+XSN(M) group could significantly increase the HF, with a statistical difference (P < 0.01). The LF / HF(%) in the AF group rats was significantly higher than that in the control group rats, with a statistical difference (P < 0.001); the AF+XSN(M) group could significantly reduce the LF / HF(%), with a statistical difference (P < 0.01).

[0123] 5.3.3 Xinsuning preparations can significantly reduce sympathetic nerve activity and significantly increase vagus nerve activity.

[0124] To further investigate the role of autonomic neurons in the treatment of vagal AF with the xinsuning preparation, peripheral autonomic nerve recordings were performed to analyze the cervical sympathetic and vagal nerve activities in rats, respectively.

[0125] like Figure 7 , Figure 8 and Fig. 9 As shown in the figure, compared with the control group, the vagus nerve activity of the rats in the AF group was significantly decreased, with a statistical difference (P < 0.01); the vagus nerve activity of the AF+XSN(M) group was increased, with a statistical difference (P < 0.05). Compared with the control group, the sympathetic nerve activity of the rats in the AF group was increased, with a statistical difference (P < 0.05); the sympathetic nerve activity of the rats in the AF+XSN(M) group was significantly decreased, with a statistical difference (P < 0.001).

[0126] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. Use of a xinsuning preparation in preparing a drug for preventing or treating vagal atrial fibrillation.

2. The use of the Xinsuning preparation in the preparation of drugs for preventing or treating vagal atrial fibrillation by reducing the incidence of vagal atrial fibrillation.

3. Use of the Xinsuning preparation in the preparation of drugs for preventing or treating vagal atrial fibrillation by shortening the duration of vagal atrial fibrillation.

4. Use of the Xinsuning preparation in the preparation of drugs for preventing or treating vagal atrial fibrillation by reducing the P wave duration.

5. Use of the Xinsuning preparation in the preparation of drugs for preventing or treating vagal atrial fibrillation by reducing sympathetic nerve tension and sympathetic nerve activity.

6. Use of the Xinsuning preparation in preparing drugs for preventing or treating vagal atrial fibrillation by increasing vagal nerve tone and vagal nerve activity.

7. The use according to any one of claims 1 to 6, characterized in that: The dosage of the Xinsuning preparation for preventing or treating vagal atrial fibrillation is 0.041 g / kg.d to 0.165 g / kg.d.

8. The use according to any one of claims 1 to 6, characterized in that: The Xinsuning preparation is prepared by combining the following raw materials in proportion by mass: the mass ratio of coptis chinensis, pinellia ternata, poria, immature bitter orange, radix chebulae, lotus seed core, sophora flavescens, artemisia annua, ginseng, ophiopogon japonicus and liquorice is 1:0.745:0.745:0.496:0.745:0.135:0.745:0.745:0.496:0.745:0.

496.

9. The use according to claim 8, characterized in that The Xinsuning preparation is prepared by the following steps: (1) Purify, slice, and dry the raw materials of coptis root, pinellia tuber, tuckahoe, immature bitter orange, radix chebulae, lotus seed core, sophora flavescens, artemisia annua, ginseng, ophiopogon japonicus, and liquorice according to the mass ratio; (2) Take coptis root, immature bitter orange, radix chebulae, lotus seed core and sophora flavescens, add 60% ethanol, reflux extraction twice, each time for 1.5 hours, combine the extracts, filter, reduce the pressure of the filtrate to recover ethanol, continue to concentrate to a thick paste with a relative density of 1.36 to 1.38 at 60°C, dry at 80°C to form a dry paste, and discard the residue; (3) Take ginseng, Poria cocos, and Pinellia tuber and add 70% ethanol, reflux extract twice, each time for 2 hours, combine the extracts, filter, and collect the filtrate in another container for later use; set aside the residue for later use; (4) The residue obtained in step (3) is combined with Ophiopogon japonicus, Artemisia annua and Glycyrrhiza uralensis, and decocted twice with water, each time for 1 hour, the decoctions are combined, filtered, and the filtrate is concentrated to a clear paste with a relative density of 1.05 to 1.06 at 80° C., ethanol is added to make the alcohol content 70%, stirred evenly, and allowed to stand for 24 hours; the supernatant is taken, filtered, and the filtrate is combined with the filtrate collected in another device in step (3), the ethanol is recovered under reduced pressure, and the mixture is further concentrated to a thick paste with a relative density of 1.36 to 1.38 at 60° C., and dried at 80° C. to obtain a dry paste; (5) The two dry pastes obtained in step (2) and step (4) are combined, crushed into fine powder, and other pharmaceutical excipients are added, mixed, and prepared into a preparation.

10. The use according to any one of claims 1 to 6, characterized in that: The Xinsuning preparation is an oral liquid preparation such as tablets, capsules, granules, pills, etc.

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