CN113559206B

CN113559206B - Compound traditional Chinese medicine for preventing and treating atrial fibrillation

Info

Publication number: CN113559206B
Application number: CN202110941627.7A
Authority: CN
Inventors: 宫丽鸿
Original assignee: Individual
Current assignee: Guizhen Biotechnology Liaoning Co ltd
Priority date: 2021-08-17
Filing date: 2021-08-17
Publication date: 2022-03-01
Anticipated expiration: 2041-08-17
Also published as: CN113559206A

Abstract

The invention discloses a compound traditional Chinese medicine for preventing and treating atrial fibrillation, belonging to the technical field of traditional Chinese medicines; the compound Chinese medicinal materials comprise pericarpium Citri Tangerinae, rhizoma Pinelliae, caulis Bambusae in Taenia, fructus Aurantii, Scutellariae radix, bupleuri radix, radix Paeoniae alba, radix Glycyrrhizae Preparata, radix Sophorae Flavescentis, rhizoma Nardostachyos, Bombyx Batryticatus, herba Taxilli, Saviae Miltiorrhizae radix, processed Notoginseng radix, ramulus Cinnamomi, Os Draconis and Concha Ostreae; the compound traditional Chinese medicine can inhibit the occurrence and development of atrial fibrillation from multiple channels and multiple target points, can improve the activation, oxidative stress, fibrosis and prothrombotic state of autonomic nerves of the atrial fibrillation, has good treatment effect on phlegm-turbidity and blood-stasis type atrial fibrillation, can obviously improve the attack frequency and duration of the atrial fibrillation, and has the clinical effective rate as high as 86%; the recurrence rate of treating atrial fibrillation by adopting the compound traditional Chinese medicine is low, and the recurrence rate of three months is 80%; meanwhile, the compound traditional Chinese medicine provided by the invention is used for treating atrial fibrillation, and has no obvious toxic or side effect and adverse reaction.

Description

Compound traditional Chinese medicine for preventing and treating atrial fibrillation

Technical Field

The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a compound traditional Chinese medicine for preventing and treating atrial fibrillation.

Background

Atrial Fibrillation (abbreviated as "Atrial Fibrillation") is the most common cardiac arrhythmia in clinic, meaning the loss of regular, ordered electrical Atrial activity, and the replacement with a rapidly disorganized Fibrillation wave is the most serious disorder of electrical Atrial activity. Atrial chaotic fibrillation loses effective contraction and relaxation, worsening or loss of atrial pumping function, and, in addition, diminished conduction of atrioventricular junctions to rapid atrial activation, causes a very irregular ventricular response. Therefore, ventricular rhythm disorder, cardiac function impairment and atrial mural thrombosis are the main pathological characteristics of patients with atrial fibrillation, the rapid ventricular rate affects diastole, the cardiac pumping function is reduced, and the patients have cardiac insufficiency or cerebral infarction symptoms. By 2010, about 3350 million patients with atrial fibrillation are estimated, the rate and incidence of patients with atrial fibrillation gradually increase, and the atrial fibrillation aggravates the risks of heart failure and stroke, and affects the quality of life. The high incidence and prevalence of atrial fibrillation not only bring pain to patients, but also bring economic pressure to society, and is a cardiovascular epidemic that seriously harms the health of people at present.

At present, western medicine treatment for atrial fibrillation mainly comprises rhythm control (including non-medicine treatment such as anti-arrhythmia medicine treatment, cardioversion and radiofrequency ablation treatment), ventricular rate control (taking beta receptor blockers, non-dihydropyridine calcium ion antagonists, digoxin, amiodarone and the like), prevention of thromboembolism and stroke caused by atrial fibrillation, atrial fibrillation upstream treatment, atrial fibrillation surgical treatment and the like, but the medicines have more adverse reactions, can resist arrhythmia, but have side effects of arrhythmia, and western medicine treatment methods such as a commonly used radiofrequency ablation technology and the like are expensive and have high recurrence rate and can cause myocardial burn, so the current western medicine treatment for atrial fibrillation is a double-edged sword, and the western medicine treatment for atrial fibrillation is in a bottleneck stage.

With the vigorous development of the traditional Chinese medicine industry, the traditional Chinese medicine shows excellent advantages in the treatment of a plurality of diseases, such as small side effect, lasting drug effect, higher patient compliance, capability of playing a role from a plurality of ion channels and a plurality of target points, and the like, and obviously improves the clinical symptoms and prognosis of patients. Therefore, if a traditional Chinese medicine capable of effectively preventing and treating atrial fibrillation can be researched and developed, the traditional Chinese medicine has important significance for clinical treatment of atrial fibrillation.

Disclosure of Invention

In order to overcome the technical problems in the prior art, the invention provides the compound traditional Chinese medicine for preventing and treating atrial fibrillation, and the compound traditional Chinese medicine has a good effect on treating phlegm-turbidity and blood-stasis type atrial fibrillation.

In order to achieve the purpose, the invention provides the following technical scheme:

the invention provides a compound traditional Chinese medicine for preventing and treating atrial fibrillation, which comprises, by weight, 10-20 parts of dried orange peel, 5-15 parts of pinellia ternate, 5-15 parts of bamboo shavings, 5-15 parts of fructus aurantii, 5-15 parts of scutellaria baicalensis, 5-15 parts of radix bupleuri, 5-15 parts of radix paeoniae alba, 5-15 parts of honey-fried licorice root, 10-20 parts of radix sophorae flavescentis, 5-15 parts of rhizoma nardostachyos, 2-8 parts of stiff silkworm, 5-15 parts of loranthus parasiticus, 10-20 parts of salvia miltiorrhiza, 1-5 parts of cooked pseudo-ginseng, 3-9 parts of cassia twig, 20-30 parts of keel and 20-30 parts of oyster.

Further, the raw materials comprise, by weight, 15 parts of dried orange peel, 10 parts of pinellia ternate, 10 parts of bamboo shavings, 10 parts of fructus aurantii, 10 parts of scutellaria baicalensis, 10 parts of radix bupleuri, 10 parts of radix paeoniae alba, 10 parts of honey-fried licorice root, 15 parts of radix sophorae flavescentis, 10 parts of rhizoma nardostachyos, 5 parts of stiff silkworm, 10 parts of parasitic loranthus, 15 parts of salvia miltiorrhiza, 2 parts of cooked pseudo-ginseng, 6 parts of cassia twig, 25 parts of keel and 25 parts of oyster.

The invention also provides a preparation method of the compound traditional Chinese medicine for preventing and treating atrial fibrillation, which comprises the following steps: weighing the raw materials according to the parts by weight, mixing, crushing to obtain mixed traditional Chinese medicine powder, adding water with the weight 15 times that of the mixed traditional Chinese medicine powder, soaking for 30-40 minutes, and decocting for 60-70 minutes by strong fire for the first time; adding 10 times of water by weight for decocting for 40-50 minutes for the second time; adding 5 times of water by weight for decocting for 20-30 minutes for the third time; and combining the liquid medicines obtained by 3 times, filtering by gauze, and concentrating in a water bath at 100 ℃ to one third of the original volume to obtain the compound traditional Chinese medicine for preventing and treating atrial fibrillation.

Further, the amount of water added into the traditional Chinese medicine powder is 5-10 times of the weight of the powder, and the slow fire decocting time is 3-5 hours.

The theoretical basis of the formula of the invention is as follows: with the continuous improvement of living standard of people, patients are addicted to eating fat, sweet, thick and greasy food, have heavy taste, hinder spleen from generating dampness, and dampness is accumulated into phlegm, the phlegm becomes stasis for a long time, the phlegm stasis is transformed into heat and generates wind for a long time, and the wind victory causes movement, which is in accordance with the clinical characteristics of sudden and intermittent atrial fibrillation. Therefore, the invention provides the syndrome that the atrial fibrillation mechanism in the traditional Chinese medicine is phlegm-stasis wind-driving, and wind pathogen disturbs the veins of heart by phlegm-stasis.

The phlegm turbidity and blood stasis type atrial fibrillation has the clinical performance characteristics that:

the main symptoms are: palpitation, restlessness, chest distress, restlessness, easy operation due to convulsion, and heart pain like needling.

With accompanying symptoms: insomnia and dreamful sleep, dry and bitter mouth, purple lips and nails, constipation and scanty and brownish urine.

Tongue manifestation: the tongue is purple dark or has ecchymosis, and the coating is yellow and greasy.

The pulse condition: the pulse is wiry, smooth or knotted.

According to the theoretical research results and symptom expression, the compound traditional Chinese medicine for preventing and treating atrial fibrillation is researched and developed.

Compared with the prior art, the invention has the following beneficial effects:

the compound traditional Chinese medicine is prepared according to the principle of monarch, minister, assistant and guide, takes scutellaria baicalensis, caulis bambusae in taeniam, pericarpium citri reticulatae, pinellia ternate, radix sophorae flavescentis, salvia miltiorrhiza, cooked panax notoginseng and stiff silkworm as monarch medicines, and has the effects of clearing heat, drying dampness, reducing phlegm, promoting blood circulation, removing blood stasis, calming endogenous wind and dredging collaterals; pinellia ternate and dried orange peel have the effects of drying dampness and eliminating phlegm, pungent and dispersing and regulating qi, scutellaria baicalensis, caulis bambusae in taeniam and sophora flavescens have the effects of clearing heat and drying dampness, reducing phlegm and descending qi, regulating spleen and stomach to ensure that phlegm is self-dissolved, salvia miltiorrhiza and cooked panax notoginseng have the effects of nourishing blood and promoting blood circulation, removing stasis and dredging collaterals, and are matched with stiff silkworm to obtain the effects of calming endogenous wind and dredging collaterals so as to inhibit the symptom of 'wind movement'; radix bupleuri, fructus aurantii, radix paeoniae alba and herba taxilli are used as ministerial drugs to regulate qi activity and communicate heart and kidney; radix bupleuri and fructus aurantii have the functions of soothing liver, regulating qi, reducing phlegm, radix paeoniae alba has the functions of nourishing yin, softening liver, nourishing blood, inhibiting liver wind, enabling qi movement to be smooth, enabling the heart and kidney to ascend and descend normally, and enabling loranthus parasiticus to enter liver and kidney channels, so that the traditional Chinese medicine has the functions of dispelling wind and tonifying liver and kidney; cassia twig, honey-fried licorice root, dragon bone and oyster shell are used as adjuvant drugs, so that the qi is tonified, the yang is boosted, the heart vessel is warmed, and the nerves and palpitation are calmed; prepared licorice root, radix Glycyrrhizae Praeparata, ramulus Cinnamomi with pungent and sweet taste and capable of resolving yang, has the effects of invigorating qi and supporting yang, and warming and dredging heart vessels; the raw dragon bone and the raw oyster shell have the effects of tranquilizing mind and astringing floating heart, and the four medicines are compatible to achieve the effects of tonifying qi and yang, warming and dredging heart vessels, calming nerves and stabilizing palpitation and help to recover the function of the sinoatrial node; the nardostachys chinensis batal is used as a guiding drug, has the effects of promoting qi circulation, relieving depression, activating spleen, invigorating stomach and promoting qi circulation to reduce phlegm. The medicines in the formula are combined to play the effects of clearing heat, eliminating phlegm, removing blood stasis, calming endogenous wind, stopping palpitation and soothing nerves. The compound traditional Chinese medicine has good treatment effect on the phlegm-turbidity and blood-stasis type atrial fibrillation, can obviously improve the attack frequency and duration of the atrial fibrillation, and has the clinical effective rate as high as 86%; the recurrence rate of treating atrial fibrillation by adopting the compound traditional Chinese medicine is low, and the recurrence rate of three months is 80%; meanwhile, the compound traditional Chinese medicine provided by the invention is used for treating atrial fibrillation, and has no obvious toxic or side effect and adverse reaction.

The invention adopts the compound traditional Chinese medicine to treat the atrial fibrillation, and reduces the economic burden of patients and the national medical cost compared with non-medicine treatment means such as radio frequency ablation, left atrial appendage occlusion and the like. In the action mechanism, the compound traditional Chinese medicine can inhibit the occurrence and development of atrial fibrillation from multiple paths and multiple targets, greatly shortens the duration time of the atrial fibrillation, can improve the activation, oxidative stress, fibrosis and prothrombotic state of autonomic nerves of the atrial fibrillation, solves the clinical problems, widens the possibility of the current atrial fibrillation treatment, further promotes the vigorous development of the traditional Chinese medicine in the atrial fibrillation treatment, and has great social value.

Drawings

FIG. 1 is an electrocardiogram of rats before modeling;

FIG. 2 is an electrocardiogram of atrial fibrillation in rats;

FIGS. 3(a) to (f) are graphs showing the myocardial ultrastructures of rats of Control group, Model group, High dose group, Middle dose group, Low dose group and Verapamil group, respectively.

Detailed Description

Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention. It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

As used herein, the terms "comprising," "including," "having," "containing," and the like are open-ended terms that mean including, but not limited to.

Example 1

The compound traditional Chinese medicine for preventing and treating atrial fibrillation is prepared from the following raw materials:

15g of dried orange peel, 10g of pinellia ternate, 10g of bamboo shavings, 10g of fructus aurantii, 10g of scutellaria baicalensis, 10g of radix bupleuri, 10g of radix paeoniae alba, 10g of honey-fried licorice root, 15g of radix sophorae flavescentis, 10g of rhizoma nardostachyos, 5g of stiff silkworm, 10g of parasitic loranthus, 15g of salvia miltiorrhiza, 2g of cooked pseudo-ginseng, 6g of cassia twig, 25g of dragon bone and 25g of oyster.

The preparation method comprises the following steps:

weighing decoction pieces of the raw materials according to the weight, mixing, crushing to obtain mixed traditional Chinese medicine powder, adding 15 times of water by weight, soaking for 40 minutes, decocting for 70 minutes by strong fire, and pouring out liquid medicine; then adding 10 times of water by weight, decocting for 50 minutes, and pouring out the liquid medicine; then adding 5 times of water by weight, decocting for 30 minutes, and pouring out the liquid medicine; and combining the liquid medicines obtained by 3 times, filtering by gauze, and concentrating in a water bath at 100 ℃ to one third of the original volume to obtain the compound traditional Chinese medicine for preventing and treating atrial fibrillation.

Example 2

10g of dried orange peel, 5g of pinellia ternate, 15g of bamboo shavings, 5g of fructus aurantii, 15g of scutellaria baicalensis, 5g of radix bupleuri, 15g of radix paeoniae alba, 5g of honey-fried licorice root, 20g of radix sophorae flavescentis, 5g of rhizoma nardostachyos, 8g of stiff silkworm, 5g of parasitic loranthus, 20g of salvia miltiorrhiza, 1g of cooked pseudo-ginseng, 9g of cassia twig, 20g of dragon bone and 30g of oyster.

The preparation method comprises the following steps:

weighing decoction pieces of the raw materials according to the weight, mixing, crushing to obtain mixed traditional Chinese medicine powder, adding 15 times of water by weight, soaking for 30 minutes, decocting for 60 minutes by strong fire, and pouring out liquid medicine; then adding 10 times of water by weight, decocting for 40 minutes, and pouring out the liquid medicine; then adding 5 times of water by weight, decocting for 30 minutes, and pouring out the liquid medicine; and combining the liquid medicines obtained by 3 times, filtering by gauze, and concentrating in a water bath at 100 ℃ to one third of the original volume to obtain the compound traditional Chinese medicine for preventing and treating atrial fibrillation.

Example 3

20g of dried orange peel, 15g of pinellia ternate, 5g of bamboo shavings, 15g of fructus aurantii, 5g of scutellaria baicalensis, 15g of radix bupleuri, 5g of radix paeoniae alba, 15g of honey-fried licorice root, 10g of radix sophorae flavescentis, 15g of rhizoma nardostachyos, 2g of stiff silkworm, 15g of parasitic loranthus, 10g of salvia miltiorrhiza, 5g of cooked pseudo-ginseng, 3g of cassia twig, 30g of dragon bone and 20g of oyster.

The preparation method comprises the following steps:

weighing decoction pieces of the raw materials according to the weight, mixing, crushing to obtain mixed traditional Chinese medicine powder, adding 15 times of water by weight, soaking for 40 minutes, decocting for 70 minutes by strong fire, and pouring out liquid medicine; then adding 10 times of water by weight, decocting for 40 minutes, and pouring out the liquid medicine; then adding 5 times of water by weight, decocting for 20 minutes, and pouring out the liquid medicine; and combining the liquid medicines obtained by 3 times, filtering by gauze, and concentrating in a water bath at 100 ℃ to one third of the original volume to obtain the compound traditional Chinese medicine for preventing and treating atrial fibrillation.

Comparative example 1

The difference from example 1 is that Bombyx Batryticatus is not included in the starting material.

Comparative example 2

The difference from example 1 is that cooked notoginseng is not included in the raw materials.

Effect verification

1. Animal experiments

1.1 Experimental animals

60 male SPF-grade SD rats (purchased from Liaoning Biotechnology GmbH, Inc., animal qualification No.: SYXK (Liao) 2013-. The experimental animals are raised in the animal experiment center of Liaoning Chinese medicine university, are fed with free drinking water and high-fat feed, and have room temperature controlled at 20-24 deg.C and humidity of about 22%.

1.2 drugs and reagents

The compound traditional Chinese medicine comprises the following components: the compound traditional Chinese medicine decoction is prepared according to the method of the embodiment 1, and is packaged in bags of 100 mL/bag and stored in a refrigerator at the temperature of-4 ℃.

Verapamil hydrochloride tablets (Tianjin central pharmaceutical limited, national standard H12020051), specification: 40 mg/tablet).

Ach (Ach) (Source leaf Biotech Co., Ltd., S30170-5g, CAS #60-31-1, molecular formula C)₇H₁₆ClNO₂) (ii) a Anhydrous calcium chloride (CaCl)₂S24110-500g, CAS # 10043-52-4); 10% chloral hydrate solution (Feijing Biotech Co., Ltd., specification: 100 mL). Rat neuropeptide Y (NPY) ELISA kit, rat superoxide dismutase (SOD) ELISA kit, rat glutathione peroxidase (GSH-Px) ELISA kit, rat matrix metalloproteinase 2(MMP-2) ELISA kit and rat P Selectin (P-Selectin) kit.

1.3 animal modeling and electrocardiographic measurements

The 60 male SD rats are randomly divided into a blank group (10 rats) and a modeling group (50 rats), and the modeling of the experiment refers to a modeling method in Chenchunlin 'establishment of rat atrial fibrillation model', and Ach (66ug/ml) -CaCl is injected into the tail vein of the rat₂(10mg/mL) of the mixture was used to prepare a turbid phlegm and blood stasis type atrial fibrillation rat model, and the dosage was calculated as 0.1mL/100g, and the solution was injected continuously for 7 days.

At 7d, all rats were weighed and anesthetized with chloral hydrate at a dose of 0.3mLl/100 g. Connecting a Power Lab 15T multi-conductive physiological recorder, lying the rat on a wood board in a supine position, fixing four limbs by rubber bands, inserting a metal needle into the skin of the disinfected four limbs, dipping a little saline water at the tail end of the lead to clamp the exposed end of the needle, measuring II-lead electrocardiogram, dynamically observing for 5 minutes, and finding that all rats of the molding group are successfully molded.

1.4 animal grouping with drug intervention

The 50 rats successfully modelled were randomly divided into 6 groups, i.e., 10 Model groups, 10 High-dose groups, 10 medium-dose groups, 10 Low-dose groups, 10 Verapamil groups, and 6 Control groups, i.e., blank groups (Control groups) without modeling.

The rat administration dosage is converted by referring to the coefficient of equivalent dose ratio (1:6.25) converted according to body surface area in pharmacological experiment methodology compiled by Xutaiyun master, the weight of a human is 60kg as reference, and the specific intragastric administration dosage is as follows:

and (5) after the molding is successful, performing intragastric administration at the 2 nd day, wherein the intragastric administration volume is 3mL, and continuously performing for 2 weeks. In the period, the tail vein injection of Ach-CaCl is continued₂The mixture was administered after 1h of intragastric administration.

1.5 Observation index

After the gastric perfusion is finished, the Control group, the Model group, the High dose group, the Middle dose group and the Low dose group are respectively used for detecting the related indexes, wherein 8 rats respectively remain in the Control group, the Model group, the High dose group, the Middle dose group and the Low dose group, the Verapami group has a large number of deaths, and 7 rats remain in the Verapami group.

(1) Duration of electrocardiogram atrial fibrillation of rats in each group: the appearance of f waves and the disappearance of P waves in the electrocardiogram are used, the absolute difference of P-R intervals is used as a mark for the occurrence of atrial fibrillation, the appearance of P waves and the disappearance of f waves are used as marks for the termination of atrial fibrillation, corresponding time is recorded, and the electrocardiogram before modeling and the electrocardiogram during atrial fibrillation are respectively shown in fig. 1 and fig. 2.

(2) The change of the ultrastructure of each group of rats was observed by an ultramicroelectron microscope (x 10000 times): after the chest is opened and the heart is exposed, a liquid transfer gun is used for extracting glutaraldehyde stationary liquid, the glutaraldehyde stationary liquid is dripped onto living tissues, the heart is sheared and placed on a glass slide with the 4 ℃ glutaraldehyde stationary liquid, then a hard blade is used for shearing small tissues at the left atrium part, the small tissues are sheared into small blocks of about 1 cubic millimeter, the small blocks are placed into an EP tube containing 1mL of 4 ℃ glutaraldehyde stationary liquid, the sample is ensured to completely enter the stationary liquid, and the fixed blocks are placed in a 4 ℃ refrigerator for temporary placement.

(3) And (3) serum detection: taking blood from abdominal aorta, centrifuging for 15min, collecting supernatant, detecting the contents of NPY, SOD, GSH-Px, MMP-2 and Selectin-P in serum according to the ELISA kit specification, and operating according to the kit specification.

1.6 statistical methods

Analysis was performed using SPSS23.0 statistical software for all measurements

Expressed, ANOVA analysis of variance, P<At 0.05, P is statistically significant<0.01 is a significant difference.

1.7 results

1.7.1 general status of rats in each group

Except for the Control group, the rats in all groups had different degrees of weight loss, fair activity, sensitive response, mental fatigue, lusterless hair and inconsistent lodging. The abdominal respiration is accelerated after tail vein injection by a Model group, the heart rate is accelerated, the eyes are congested, the action is slow, the patient can recover in 10-15min, the patient is weak, sleepy, anorexia and weight loss; the High dose group and the Middle dose group have the advantages that the eyeball bleeding is reduced compared with the Model group, the activity is relatively High, and the appetite is not obviously reduced; the appetite of the Low dose group and the Verapamil group is slightly reduced, the state is hypodynamia, the eyeball still bleeds blood, and the recovery is about 4-5 min.

1.7.2 improvement of duration of atrial fibrillation of Electrocardiogram in rats after gastric lavage

All the model groups have atrial fibrillation, and have statistical significance difference (P is less than 0.05) compared with the Control group; the duration of atrial fibrillation of rats in all the drug groups is reduced to different degrees compared with that of the Model group, the difference has statistical significance (P is less than 0.05), and the comparison between the groups has no significant difference (P is more than 0.05), and the specific atrial fibrillation duration of each group is shown in a table 1:

TABLE 1 comparison of atrial fibrillation times in rats of each group

Group of	n	Duration (S)
			Control group	8	0.00
Model group	8	14.04±6.88^*
			High dose group	8	6.64±3.74^##
Middle dose group	8	6.93±1.72^##
			Low dose group	8	7.15±3.81^##
Verapamil group	7	9.02±0.81^#

Note: p compared to Control group<0.05，**P<0.01; compared with a Model group^#P<0.05，^##P<0.01。

1.7.3 Observation of atrial muscle ultrastructures of rats in each group, the myocardial ultrastructures of rats in Control group, Model group, High dose group, Middle dose group, Low dose group and Verapaml group are shown in FIGS. 3(a) to (f), and it can be seen from FIG. 3 that: the muscle segments of the atrial myocytes of rats in the Control group are complete and clear, the arrangement of the Z bands is neat, coherent, clear and orderly, the I bands are clear and visible, the arrangement of myofilaments is neat and compact, the mitochondrial structure is normal, the arrangement of mitochondrial cristae is compact, the structure is complete, and the phenomenon of intercalated disc separation is not seen.

Model group rats had broken muscle segments of atrial myocytes, discontinuous Z-bands, disordered arrangement, blurred I-bands, myofilament lysis and breakage of the cardiomyocytes, voids, mitochondrial swelling, increase and even rupture, and cristae lysis and aggregation together in vacuole-like changes.

The muscle segments of the atrial myocytes of the rats in the High dose group are slightly broken and relatively regularly arranged, myofilaments are slightly dissolved, mitochondria still swell and increase, but cristae dissolution is obviously reduced, the structure is relatively complete, and the phenomenon of intercalated disc separation is improved.

The muscle segments of atrial myocytes of rats in the Middle dose group are relatively complete, and the Z has a dislocation phenomenon, is orderly arranged, slightly blurs myofilaments, swells mitochondria, occasionally dissolves mitochondria and has relatively complete structure.

The muscle segments of atrial myocytes of rats in the Low dose group are fractured, the Z bands are discontinuous and are not regularly arranged, the I bands are unclear, the mitochondria are swollen and increased, the ridge is fractured and dissolved, and the individual structures are incomplete.

The muscle segments of atrial myocytes of rats in the Verapamil group are slightly fractured, the Z bands are widened, interrupted and discontinuous, the arrangement is relatively regular, the I bands are slightly fuzzy, mitochondria are increased and gathered together, and partial cristae is fractured and dissolved.

1.7.4 effects of autonomic nerves

The experimental results show that: compared with the Control group, the serum NPY level of the rats in the Model group is obviously increased (P is less than 0.01), the serum NPY level of the rats in the rest groups except the Low dose group is obviously reduced after the drug is taken (P is less than 0.01), no obvious difference exists in comparison between the groups (P is more than 0.05), and the serum neuropeptide Y level of each group of rats is shown in a table 2:

TABLE 2 serum neuropeptide Y levels in various groups of rats

Group of	n	NYP(ng/mL)
			Control group	8	5.20±0.51
Model group	8	8.00±0.49^**
			High dose group	8	7.28±0.62^##
Middle dose group	8	6.02±0.2^##
			Low dose group	8	7.81±0.91
Verapamil group	7	6.01±0.11^##

Note: p compared to Control group<0.05，^**P<0.01(ii) a Compared with a Model group^#P<0.05，^##P<0.01。

The serum NPY level of rats in the Model group is obviously increased compared with that of rats in the Control group (P <0.01), which represents that sympathetic nerves are activated when atrial fibrillation occurs, and the serum NPY level of rats in all the drug groups except the Low dose group is reduced (P < 0.01). The compound traditional Chinese medicine of the invention is proved to be capable of inhibiting sympathetic nerve activation, thereby delaying the development of atrial fibrillation.

1.7.5 oxidative stress related index

The experimental results show that: compared with the Control group, the serum GSH-Px and SOD level of the Model group rats are obviously reduced (P is less than 0.01). Compared with the Model group, the serum SOD levels of the High dose group, the Middle dose group and the Verapamide group are obviously increased (P <0.05), and no obvious difference exists in comparison among the groups (P > 0.05); for GSH-Px, compared with the Model group, the serum GSH-Px level of all the drug groups is obviously increased (P is less than 0.05), the serum GSH-Px level of the middle-high, middle-Low dose group and the serum GSH-Px level of the Verapamide group are not obviously different (P is more than 0.05), the oxidative stress index SOD and the GSH-Px level of each group of rats are shown in a table 3:

TABLE 3 oxidative stress indexes SOD, GSH-Px for each group of rats

Group of	n	SOD(ng/mL)	GSH-Px(ng/mL)
				Control group	8	2.45±0.19	3.56±0.28
ModelGroup of	8	2.12±0.21^**	3.27±0.12^**
				High dose group	8	2.40±0.26^##	3.61±0.06^##
Middle dose group	8	2.40±0.12^##	3.60±0.08^##
				Low dose group	8	2.27±0.23	3.47±0.15^#
Verapamil group	7	2.36±0.07^#	3.51±0.19^#

1.7.6 fibrosis and prothrombotic state detection

The experimental results show that: MMP-2: compared with the Control group, the serum MMP-2 level of the Model group is increased (P is less than 0.05), and the difference has statistical significance. Compared with the Model group, the serum MMP-2 level of the High dose group, the Middle dose group and the Verapamide group is obviously reduced (P is less than 0.01), the comparison between the groups has no statistical significance (P is more than 0.05), and the fibrosis index MMP-2 level of rats in each group is shown in a table 4.

selectin-P: compared with the Control group, the serum Selectin-P level of the Model group is obviously increased (P <0.01), the serum Selectin-P level of the rest groups except the Verapamide group is obviously reduced (P <0.01) compared with the Model group after the drug is taken, the serum Selectin-P level of the Middle group is reduced more obviously (P <0.05) compared with the High group, and the level of the prothrombotic state Selectin-P of each group of rats is shown in a table 4:

TABLE 4 fibrosis indices MMP-2 and prothrombotic State Selectin-P

Group of	n	MMP-2(ng/mL)	Selectin-P(pg/mL)
				Control group	8	247.07±60.00	26.81±3.49
Model group	8	300.33±52.61^*	44.83±6.91^**
				High dose group	8	257.4±35.06^#	28.33±8.76^##
Middle dose group	8	258.89±11.08^#	19.04±4.14^##
				Low dose group	8	293.19±38.73	27.22±2.95^##
Verapamil group	7	258.12±13.40^#	34.72±12.01

Note: p compared to Control group<0.05，^**P<0.01; compared with a Model group^#P<0.05，^##P<0.01。

Oxidative stress plays an important role in the development and maintenance of atrial fibrillation, in the molding process, in Ach-CaCl₂Under the action of the mixture, oxidative stress occurs, oxygen free radicals are greatly increased, and SOD and GSH-Px are greatly consumed in the oxidation resisting process, so that the contents of SOD and GSH-Px in the serum of Model group rats are reduced (P)<0.01) and simultaneously initiates tissue fibrosis, which leads to the elevation of MMP-2. The serum GSH-Px level of all the medicinal groups of rats is obviously increased (P is less than 0.05) compared with that of a model; for SOD, the serum SOD level of rats of only High dose group, Middle dose group and Verapamil group is obviously increased, and the difference has statistical significance (P)<0.05); for MMP-2, the serum MMP-2 levels were significantly reduced in the High dose, Middle dose and Verapamil groups (P < 0.05). The compound traditional Chinese medicine can inhibit oxidative stress, thereby improving the fibrosis degree caused by atrial fibrillation.

selectin-P is also known as GMP-140, CD 62P. Is a transmembrane glycoprotein with the molecular weight of 140KD, the increase of selectin-P during AF plays an important role in the formation of atrial fibrillation thrombus, and the thrombospondin-P mediates the adhesion of platelets to endothelial cells and the connection of the platelets and neutrophils, activates the neutrophils, releases vasoactive substances and oxygen metabolites, causes vasoconstriction, blocks small blood vessels and starts the thrombus formation process. The serum Selectin-P content of rats in the Model group is increased (P <0.01), and the serum Selectin-P levels of rats in the High dose group, the Middle dose group and the Low dose group are reduced to different degrees after the drug is applied.

Therefore, the compound traditional Chinese medicine can improve the autonomic nerve activation, oxidative stress, fibrosis degree and prothrombotic state of atrial fibrillation, thereby delaying the development of atrial fibrillation and improving clinical symptoms.

2. Clinical validation

2.1 general data

The study subjects selected 2019.6-2021.6 cases of paroxysmal turbid phlegm and blood stasis type atrial fibrillation patients with clear diagnosis in outpatient service of subsidiary hospitals of Liaoning traditional Chinese medicine university, and the age range of the patients is 48-87 years old.

Diagnosis standard of phlegm-turbidity and blood-stasis type atrial fibrillation patient

Western diagnostic criteria: according to old-fashioned book of Huangwang clinical electrocardiography (6 th edition), the diagnosis of atrial fibrillation for type classification refers to the diagnosis and treatment guideline for atrial fibrillation published by the European Heart disease society (ESC) in 2016.

The traditional Chinese medicine diagnosis standard is as follows: the diagnosis of palpitation is referred to "internal science of traditional Chinese medicine"; dialectical typing diagnosis of traditional Chinese medicine: type of turbid phlegm and blood stasis: reference is made to GB "syndrome part of Chinese medicine clinical diagnosis and treatment terms" 2002 edition "guide principles of clinical research on Chinese medicines (new drugs)".

And (3) master certificate: palpitation, asthenia, shortness of breath, chest distress.

Secondary verification: anorexia, nausea, emesis, limb sleepiness, dizziness, dysphoria, irascibility, insomnia, dry mouth, bitter taste, yellow and red urine, constipation, dark tongue, ecchymosis, yellow and greasy tongue coating, and wiry and slippery pulse.

Inclusion criteria were:

(1) the diagnosis standard of the paroxysmal atrial fibrillation in western medicine is met.

(2) The diagnostic standard of the traditional Chinese medicine for palpitation, turbid phlegm and blood stasis is met.

(3) The age is greater than 18 years.

(4) The attack frequency is more than or equal to 2 times per month.

(5) The conventional western medicine oral administration does not obviously relieve the relapse author or relapse after the radiofrequency ablation.

(6) There are no contraindications for treatment, such as sick sinus syndrome, II °, III ° AVB.

(7) Agreeing to oral Chinese medicinal treatment.

Exclusion criteria:

(1) atrial fibrillation with reversible etiology;

(2) has serious primary diseases of liver, kidney, hemopoietic system, endocrine system, etc., or diseases affecting its survival, such as tumor, AIDS, etc.;

(3) acute infection, rheumatic heart disease, acute coronary syndrome, hypertension and acute and severe diseases occur during observation;

(4) are participating in other drug clinical trials;

(5) those with allergic constitution;

(6) pregnant or lactating women.

Standard of case elimination

(1) The patients have special physiological changes of certain complications and complications during taking the medicine, and are not suitable for continuously taking the traditional Chinese medicine orally.

(2) The patient has incomplete data entry, which affects the validity judgment.

Standard of detachment

(1) Patients who were given up treatment or missed visits did not complete a 12-week clinical visit.

(2) Patient compliance was poor and the drug was not taken at the prescribed dose and frequency.

200 patients selected according to the above criteria were randomly divided into groups A to D4 of 50 patients each. General data for four groups of patients were comparable (P > 0.05).

2.2 treatment regimens

Group A: amiodarone hydrochloride tablets (coda) are orally taken, twice in the morning and at night, half an hour after meals.

Group B: the compound traditional Chinese medicine decoction prepared in the embodiment 1 is orally taken, the dosage is 100 mL/time, 3 times a day in the morning, in the middle of the day and at night, the decoction is taken half an hour after meals, and 12 weeks are a large treatment course.

Group C: the compound traditional Chinese medicine decoction prepared in the comparative example 1 is orally taken, the dosage is 100 mL/time, 3 times a day in the morning, in the middle of the day and at night, the decoction is taken half an hour after meals, and 12 weeks are a large course of treatment.

Group D: the compound traditional Chinese medicine decoction prepared in the comparative example 2 is orally taken, the dosage is 100 mL/time, 3 times a day in the morning, in the middle of the day and at night, the decoction is taken half an hour after meals, and 12 weeks are a large treatment course.

Patients with other basic diseases continue to take conventional western medicines orally, and patients with the traditional Chinese medicine treatment group can take the beta receptor blocker temporarily if the onset of the rapid arrhythmia occurs. During the treatment period, the low-salt and low-fat diet avoids taking pungent beverages such as strong wine, coffee, concentrated tea and the like, avoids spicy, greasy, cold and raw products, and avoids strenuous exercise.

2.3 Observation of indicators

2.3.1 determination of efficacy

(1) The evaluation standard of traditional Chinese medicine syndrome curative effect refers to 2002 edition of Chinese medicine new drug clinical research guiding principles, which is shown in Table 5:

TABLE 5 evaluation criteria for efficacy of traditional Chinese medicine syndromes

Show effect	The clinical symptoms and physical signs are obviously improved, and the syndrome integral is reduced by more than or equal to 70 percent.
		Is effective	The clinical symptoms and physical signs are improved, and the syndrome integral is reduced by 30 to 70 percent.
Invalidation	Clinical symptomsNo obvious improvement or even aggravation of the symptoms and signs and reduction of the syndrome score<30％
		Weighting device	The clinical symptoms and physical signs are aggravated, and the syndrome score is reduced<0。

Integrated effective rate (%) (total integration before treatment-total integration after treatment)/total integration before treatment × 100%.

(2) The score of the main traditional Chinese medicine symptoms improves the condition.

(3) Atrial fibrillation attack frequency and duration.

(4) The maintenance rates of the patients in each group for three months of treatment were compared.

2.3.2 safety evaluation of the occurrence of obvious adverse reactions after administration, liver and kidney functions and hematuria.

2.4. Statistical method

Data were analyzed using SPSS23.0 statistical software. Average +/-standard deviation for measuring data "

Means that the pre-and post-treatment comparisons within the group were tested with paired t-tests, and the pre-and post-treatment comparisons between the groups were tested with independent samples t-tests; comparing the grade data with a rank sum test; checking the counting data with a chi-square with P<0.05 is statistically different, expressed as P<0.01 is a significant difference.

2.5 results

2.5.1 comparison of the therapeutic effects of the syndromes of the patients

Group B, the treatment with the traditional Chinese medicine of example 1, the significant efficiency is 48%, and the total effective rate is 86% (43/50); the obvious effect rate of the group A, namely the amiodarone hydrochloride tablet treatment group is 18%, the total effective rate is 46% (23/50), the difference of the traditional Chinese medicine syndrome curative effect of each group has statistical significance (P is less than 0.05) through the rank sum test, and the comparison result of the traditional Chinese medicine syndrome curative effect of each group is shown in the table 6:

TABLE 6 comparison of the therapeutic effects of the syndromes in each group

Group of

Example number (n)

Show effect

Is effective

Invalidation

Weighting device

Total effective rate (%)

Group A

50 examples of

9(18％)

14(28％)

27(54％)

0

46

Group B

50 examples of

24(48％)

19(38％)

7(14％)

0

86

Group C

50 examples of

20(40％)

17(34％)

13(26％)

0

74

Group D

50 examples of

18(36％)

20(40％)

12(24％)

0

76

2.5.2 comparison of scores of the symptoms of the main TCM drugs before and after each group of treatment

The difference of the traditional Chinese medicine symptom scores of the four groups before treatment has no statistical significance (P is more than 0.05), and the traditional Chinese medicine composition has comparability. The score of the main symptoms of the four groups after treatment is reduced (P is less than 0.05) compared with that before treatment, the traditional Chinese medicine syndrome of the group B is improved more remarkably (P is less than 0.05) compared with that of the group A, and the score comparison results of the main symptoms before and after treatment of each group are shown in a table 7:

TABLE 7 comparison of chief complaint scores before and after treatment for each group

Note that comparison with this group before treatment^*P＜0.05，^**P is less than 0.01; comparison with group A^#P＜0.05，^##P＜0.01。

2.5.3 comparison of atrial fibrillation attack frequency before and after treatment of groups

Before treatment, the difference of the atrial fibrillation attack frequency of each group has no statistical significance (P is more than 0.05), and the atrial fibrillation attack frequency is comparable. After treatment, the atrial fibrillation attack frequency of each group is obviously reduced compared with that before treatment, the difference has statistical significance (P is less than 0.01), the attack frequency of the group B is more obviously reduced compared with that of the group A, the difference has statistical significance (P is less than 0.01), and the comparison result of the atrial fibrillation attack frequency before and after treatment of each group is shown in a table 8:

TABLE 8 comparison of mean frequency of onset of atrial fibrillation before and after treatment in each group

Note that comparison with this group before treatment^*P＜0.05，^＊＊P is less than 0.01; comparison with group A^#P＜0.05，^##P＜0.01。

2.5.4 comparison of atrial fibrillation duration before and after treatment in each group

Before treatment, the difference of the atrial fibrillation duration time of each group has no statistical significance (P is more than 0.05), and the atrial fibrillation duration time has comparability; after treatment, the atrial fibrillation duration time of each group is obviously reduced compared with that before treatment, the difference has statistical significance (P is less than 0.01), the duration time of the group B is reduced more obviously than that of the group A, the difference has statistical significance (P is less than 0.01), and the comparison result of the atrial fibrillation duration time before and after treatment of each group is shown in a table 9:

TABLE 9 comparison of mean duration of atrial fibrillation before and after treatment in each group

Group of	Example number (n)	Before treatment/h	After treatment/h
				Group A	50 examples of	8.16±4.61	1.41±0.88^**
Group B	50 examples of	8.54±3.49	0.53±0.19^**##
				Group C	50 examples of	8.47±2.28	0.73±0.15
Group D	50 examples of	8.50±1.66	0.61±0.09

Note that P <0.05 compared to the group before treatment,^＊＊p is less than 0.01; comparison with group A^#P＜0.05，^##P＜0.01。

2.5.5 comparison of maintenance rates of three-month-to-three-month treatment in each group

The three-month-transition maintenance rate of patients in group B is 80%, the three-month-transition maintenance rate of patients in group A is 38%, the three-month-transition maintenance of patients in each group is different, the effect of group B is superior to that of group A (P is less than 0.01), and the comparison result of the three-month-transition maintenance rate of patients in each group is shown in a table 10:

TABLE 10 comparison of three-month-to-month-transition maintenance rates for each group

2.5.6 safety evaluation of patients in each group have no adverse reaction, regular hematuria and no abnormality of liver and kidney function during three months of administration.

Animal experiments and clinical verification are carried out by adopting the compound traditional Chinese medicines prepared in the embodiments 2 and 3 according to the method, and the obtained effect is equivalent to the verification result of the compound traditional Chinese medicine in the embodiment 1.

The above description is only for the preferred embodiment of the present invention, and the protection scope of the present invention is not limited thereto, and any person skilled in the art should be able to cover the technical scope of the present invention, the technical solution and the inventive concept of the present invention equivalent or change within the technical scope of the present invention.

Claims

1. The compound traditional Chinese medicine for preventing and treating phlegm-turbidity and blood-stasis type atrial fibrillation is characterized by being prepared from the following components in parts by weight: 10-20 parts of dried orange peel, 5-15 parts of pinellia ternate, 5-15 parts of bamboo shavings, 5-15 parts of fructus aurantii, 5-15 parts of scutellaria baicalensis, 5-15 parts of radix bupleuri, 5-15 parts of radix paeoniae alba, 5-15 parts of honey-fried licorice root, 10-20 parts of radix sophorae flavescentis, 5-15 parts of nardostachyos root and rhizome, 2-8 parts of stiff silkworm, 5-15 parts of loranthus parasiticus, 10-20 parts of salvia miltiorrhiza, 1-5 parts of cooked panax notoginseng, 3-9 parts of cassia twig, 20-30 parts of dragon bone and 20-30 parts of oyster.

2. The compound traditional Chinese medicine for preventing and treating phlegm-turbidity and blood-stasis type atrial fibrillation according to claim 1, is characterized by being prepared from the following components in parts by weight: 15 parts of dried orange peel, 10 parts of pinellia ternate, 10 parts of bamboo shavings, 10 parts of bitter orange, 10 parts of scutellaria baicalensis, 10 parts of radix bupleuri, 10 parts of white paeony root, 10 parts of honey-fried licorice root, 15 parts of radix sophorae flavescentis, 10 parts of nardostachys root, 5 parts of stiff silkworm, 10 parts of parasitic loranthus, 15 parts of salvia miltiorrhiza, 2 parts of cooked pseudo-ginseng, 6 parts of cassia twig, 25 parts of dragon bone and 25 parts of oyster.

3. The preparation method of the compound traditional Chinese medicine for preventing and treating phlegm-turbidity and blood-stasis type atrial fibrillation according to any one of claims 1 to 2, which is characterized by comprising the following steps of: weighing the raw materials according to the parts by weight, mixing, crushing to obtain mixed traditional Chinese medicine powder, adding water with the weight 15 times that of the mixed traditional Chinese medicine powder, soaking for 30-40 minutes, and decocting for 60-70 minutes by strong fire for the first time; adding 10 times of water by weight for decocting for 40-50 minutes for the second time; adding 5 times of water by weight for decocting for 20-30 minutes for the third time; and combining the liquid medicines obtained in 3 times, filtering by gauze, and concentrating in a water bath at 100 ℃ to one third of the original volume to obtain the compound traditional Chinese medicine for preventing and treating phlegm turbidity and blood stasis type atrial fibrillation.