Traditional Chinese medicine composition for treating heart blood stasis type arrhythmia as well as preparation method and application of traditional Chinese medicine composition

Abstract

The invention belongs to the technical field of traditional Chinese medicines, and relates to a traditional Chinese medicine composition for treating heart blood stasis type arrhythmia and a preparation method and application thereof. The traditional Chinese medicine composition is prepared from the following raw materials in parts by mass: 250 to 370 parts of radix codonopsis, 55 to 100 parts of radix notoginseng, 350 to 500 parts of rhizoma polygonati, 150 to 230 parts of rhizoma nardostachyos and 100 to 200 parts of poria cocos. The traditional Chinese medicine composition disclosed by the invention consists of five medicinal materials, namely codonopsis pilosula, pseudo-ginseng, rhizoma polygonati, rhizoma nardostachyos and poria cocos, has good effects of improving myocardial dysfunction, relieving myocardial cell injury and apoptosis and regulating cardiac troponin activity, and can be used for treating heart blood stasis type arrhythmia.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicines, and relates to a traditional Chinese medicine composition for treating cardiac blood stasis type arrhythmia, and a preparation method and application thereof. Background Art

[0002] At present, arrhythmia is still one of the public health problems that the world needs to face; about 600,000 people die of sudden cardiac death each year, of which 88% are caused by arrhythmia. With the increasing pressure of social life, the incidence of arrhythmia has gradually shown a trend of younger people; the treatment principle of arrhythmia is still to act on potassium ion channels and regulate the disorder of myocardial electrical activity. A large number of studies have shown that most of the new drugs currently on the market have pro-arrhythmic effects during antiarrhythmic treatment, and most of them have the disadvantage of negative inotropic effects.

[0003] In the treatment of arrhythmia, Chinese medicine has the characteristics of multi-target and multi-pathway synergistic effects and has unique advantages. Referring to the Chinese patent document with application number CN201510295138.3, a compound medicine for rapid supraventricular arrhythmia and its preparation method are disclosed. The compound medicine for rapid supraventricular arrhythmia is made of the following raw materials in parts by weight: 3-4 parts of scutellaria baicalensis, 3-4 parts of angelica dahurica, 6-8 parts of angelica sinensis, 1.5-2 parts of musk, 4-6 parts of capillaris, 18-25 parts of tuckahoe, 20-25 parts of cistanche deserticola, 18-22 parts of hericium erinaceus, 10-12 parts of nardostachys galanga, 1-1.5 parts of beeswax, 11-13 parts of cyperus rotundus, and 2-3 parts of chondroitin sulfate. The various ingredients in the compound medicine work together synergistically; they can effectively inhibit cell death in the patient's body, improve the patient's immunity, reduce the patient's fat and sugar content, and dilate the coronary arteries; they have outstanding therapeutic effects on the treatment of patients with arrhythmias.

[0004] Although the existing composition can be used to treat arrhythmia, since arrhythmia is a modern medical term, in clinical practice, common palpitations, chest tightness, and fatigue, shortness of breath, sweating, dizziness and other symptoms caused by insufficient blood perfusion in some patients are generally classified as "palpitations". However, traditional Chinese medicine believes that palpitations are located in the heart and are closely related to the functions of the liver, lungs, spleen, and kidneys. The essence of the disease lies in the deficiency of the root and the excess of the symptoms. The deficiency of the root is the deficiency of qi and blood, yin and yang, which is often manifested in the clinical manifestations of wind, cold, dampness, heat, or warm poison invasion syndrome; the excess of the symptoms is phlegm and blood stasis, which is often manifested in the clinical manifestations of phlegm turbidity and heart vessel stasis syndrome; therefore, according to the syndrome type of traditional Chinese medicine, arrhythmia can usually be divided into heart yang deficiency, qi and yin deficiency, heart and kidney disharmony, heart blood stasis, and phlegm and fire disturbing the heart, etc. Therefore, the treatment of arrhythmia should be combined with the syndrome type to achieve symptomatic treatment, but most of the existing arrhythmia drugs are aimed at the larger category of "palpitations" without considering specific syndrome types; and the clinical "palpitations" of traditional Chinese medicine are mostly qi and yin deficiency, heart blood stasis deficiency syndrome, and there are few existing reports on the treatment of heart blood stasis type arrhythmia, which belongs to the technical gap. Therefore, providing a composition for treating heart blood stasis type arrhythmia is a topic that needs to be developed at present. Summary of the invention

[0005] In order to solve the above technical problems existing in the background technology, the present invention provides a Chinese medicine composition for treating cardiac blood stasis type arrhythmia and a preparation method and application thereof.

[0006] The Chinese medicine composition of the present invention is composed of five medicinal materials, namely, Codonopsis pilosula, Panax notoginseng, Polygonatum sibiricum, Nardostachys grandiflora and Poria cocos, and has good effects of improving myocardial dysfunction, reducing myocardial cell damage and apoptosis, and regulating the activity of myocardial troponin, and can be used for treating arrhythmia of cardiac blood stasis type.

[0007] In order to achieve the above object, the technical solution adopted by the present invention is:

[0008] A traditional Chinese medicine composition for treating cardiac blood stasis type arrhythmia, the traditional Chinese medicine composition is composed of the following raw materials in parts by weight: 250-370 parts of Codonopsis pilosula, 55-100 parts of Panax notoginseng, 350-500 parts of Polygonatum sibiricum, 150-230 parts of Nardostachys grandiflora and 100-200 parts of Poria cocos.

[0009] It is further defined that the Chinese medicine composition is composed of the following raw materials in parts by mass: 300 parts of Codonopsis pilosula, 60 parts of Panax notoginseng, 400 parts of Polygonatum sibiricum, 200 parts of Nardostachys grandiflora and 160 parts of Poria cocos.

[0010] It is further defined that the Chinese medicine composition for treating cardiac blood stasis type arrhythmia has a significant up-regulating effect on JAK2 protein expression and MCL1 protein expression, and has a significant down-regulating effect on STAT3 protein expression and BCL2 protein expression.

[0011] The preparation method of the Chinese medicine composition for treating cardiac blood stasis type arrhythmia as described above comprises the following steps:

[0012] S1. Prepare raw materials of Codonopsis pilosula, Panax notoginseng, Polygonatum sibiricum, Nardostachys grandiflora and Poria cocos according to weight;

[0013] S2, extracting volatile oil from Nardostachys grandiflora by water extraction;

[0014] S3, extracting Panax notoginseng by ethanol reflux method to obtain Panax notoginseng filtrate and Panax notoginseng residues, concentrating the Panax notoginseng filtrate under reduced pressure to obtain Panax notoginseng paste, and decocting the Panax notoginseng residues with water to obtain Panax notoginseng decoction;

[0015] S4, after mixing Codonopsis pilosula and Polygonatum sibiricum, a decoction is obtained by a decoction method; then filtering and concentrating to obtain a Codonopsis pilosula and Polygonatum sibiricum decoction; and combining the Panax notoginseng decoction in step S3, a mixed filtrate is obtained by a water extraction and alcohol precipitation method;

[0016] S5, extracting Poria cocos by ethanol reflux method to obtain Poria cocos extract, combining it with the mixed filtrate of step S4, and concentrating under reduced pressure to obtain a thick paste;

[0017] S6, mixing the notoginseng paste of step S3 and the thick paste of step S5; then adding the nardostachys volatile oil of step S2, mixing evenly, and freeze-vacuum drying to obtain a Chinese medicine composition.

[0018] It is further defined that in step S2, 10 times the mass of water is added to the nardostachys grandiflora for extraction to obtain nardostachys grandiflora volatile oil.

[0019] It is further defined that the specific process of step S3 is:

[0020] Reflux extraction was performed twice with 10 times the mass of ethanol, each time for 2 hours, and then filtered; the two filtrates were combined and concentrated under reduced pressure to obtain Panax notoginseng paste;

[0021] Add 10 times the amount of water to the Panax notoginseng residue and boil it twice, the first time for 2 hours and the second time for 1.5 hours. The decoctions are combined to obtain the Panax notoginseng decoction.

[0022] It is further defined that the specific process of step S4 is:

[0023] Mix Codonopsis pilosula and Polygonatum sibiricum, add 10 times the weight of water and boil twice, the first time for 2 hours, the second time for 1.5 hours, and combine the two filtrates to obtain Codonopsis pilosula and Polygonatum sibiricum decoction;

[0024] The decoctions of Codonopsis pilosula and Polygonatum sibiricum and Panax notoginseng are mixed evenly and filtered, and the obtained filtrate is concentrated to a relative density of 1.20-1.30, and ethanol is added to make the alcohol content reach 65%, stirred, allowed to stand for 24 hours, filtered, and a mixed filtrate is obtained.

[0025] It is further defined that the specific process of step S5 is:

[0026] Soak Poria cocos for 12 hours, add 9 times the weight of water, heat in a water bath and reflux for extraction twice, the first time for 7.5 hours and the second time for 1.5 hours, and combine the two filtrates to obtain the Poria cocos extract;

[0027] The Poria extract is combined with the mixed filtrate of step S4 and concentrated under reduced pressure to obtain a thick paste.

[0028] The use of the Chinese medicine composition for treating cardiac blood stasis type arrhythmia in the preparation of drugs for cardiac blood stasis type arrhythmia.

[0029] The Chinese medicine composition for treating cardiac blood stasis type arrhythmia is used in the preparation of cardiac blood stasis type arrhythmia drugs by activating the JAK2-STAT3 signaling pathway to inhibit the apoptosis of myocardial cells.

[0030] The beneficial effects of the present invention are:

[0031] The Chinese medicine composition for treating arrhythmia of blood stasis type of the present invention is composed of five medicinal materials, namely, codonopsis pilosula, panax notoginseng, polygonatum sibiricum, naringa and poria. In the prescription, codonopsis pilosula is used as the main medicine, which can nourish the middle and replenish qi, calm the spirit and calm palpitations; polygonatum sibiricum is used as the assistant medicine, which can nourish yin, benefit the kidney and replenish essence. Since blood is formed under the action of yin qi, polygonatum sibiricum nourishes yin and blood while nourishing heart yin, thereby assisting codonopsis pilosula in replenishing qi and producing blood; panax notoginseng can remove blood stasis and stop bleeding, can help polygonatum sibiricum nourish heart blood, and can also help polygonatum sibiricum replenish qi, and can replenish qi and blood together with polygonatum sibiricum, and both are assistant medicines; naringa sibiricum can promote qi and relieve pain, relieve depression and wake up the spleen, so that the main and assistant medicines can replenish but not stagnate, and is the assistant medicine; poria is flat in nature, sweet and light in taste, enters the heart, lung, spleen and kidney meridians, has the effects of promoting water and dampness, invigorating spleen and calming the heart, and can transport and transform water and dampness in the prescription, help the qi of various medicines, and can directly invigorate the spleen and calm the heart, and directly reach the diseased area, so as to be the adjuvant medicine. The five medicinal materials of the present invention have the main therapeutic principles of invigorating qi and nourishing yin, promoting yang and restoring pulse, and promoting blood circulation and removing blood stasis. The various medicinal materials act synergistically, have good effects of improving myocardial dysfunction, reducing myocardial cell damage and apoptosis, and regulating cardiac troponin activity, and can be used for the treatment of arrhythmias of cardiac blood stasis type. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 A blood drug concentration-time curve of the main component of the composition;

[0033] Figure 2 The changes of ECG in lead II before and after the I / R model was established;

[0034] Figure 3 The staining images and percentage of myocardial infarction area are shown;

[0035] Figure 4 The effect of the composition on serum biochemical indexes of rats with cardiac ischemia-reperfusion injury;

[0036] Figure 5 The effects of the composition on key targets in I / R rats;

[0037] Figure 6 The pathological changes of cardiac tissues in rats with cardiac ischemia-reperfusion injury (×200);

[0038] Figure 7 The effect of the combination on the degree of myocardial fibrosis in rats with cardiac ischemia-reperfusion injury (×200);

[0039] Figure 8 and Fig. 9 The effect of the composition on the apoptosis degree of myocardial cells injured by ischemia-reperfusion in rats;

[0040] Fig.10 The principal component analysis of the metabolic profile of cardiac tissues in the blank group and the model group;

[0041] Fig.11 Volcano plot of cardiac tissue metabolites in the blank group and the model group;

[0042] Fig.12 , Fig.13 and Fig.14 Volcano plot of cardiac tissue metabolites in the drug-treated group and the model group;

[0043] Fig.15 Pathway enrichment map for differential metabolites;

[0044] Fig.16 , Fig.17 and Fig.18 The volcano plot of differentially expressed genes in the combination group-intervention I / R rats;

[0045] Fig.19 This is the GO enrichment result of differentially expressed genes in the combination group for I / R rats;

[0046] Fig. 20 This is the KEGG enrichment result of differentially expressed genes in I / R rats treated with the combination;

[0047] Fig.21 The effect of the administration of the composition on the related genes of I / R model rats;

[0048] Fig. 22 The effect of the composition on the expression of related proteins in the heart tissue of I / R model rats;

[0049] Fig.23 It is the Western Blot band diagram;

[0050] Fig.24 This is a diagram showing the mechanism of action of a Chinese herbal medicine combination in intervening in arrhythmias of the type caused by blood stasis in the heart. DETAILED DESCRIPTION

[0051] The technical solution protected by the present invention is now described in detail in conjunction with the accompanying drawings and embodiments. However, it is obvious that the described embodiments are part of the embodiments of the present application, rather than all of the embodiments. Based on the described embodiments of the present application, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present application.

[0052] The invention provides a traditional Chinese medicine composition for treating cardiac blood stasis type arrhythmia. The traditional Chinese medicine composition is composed of the following raw materials in parts by weight: 250-370 parts of codonopsis pilosula, 55-100 parts of notoginseng, 350-500 parts of polygonatum, 150-230 parts of nardostachys grandiflora and 100-200 parts of tuckahoe.

[0053] Preferably, the mass portion of Codonopsis pilosula is 250 parts, 275 parts, 290 parts, 310 parts, 330 parts, 350 parts or 370 parts; the mass portion of Panax notoginseng is 55 parts, 70 parts, 85 parts, 95 parts or 100 parts; the mass portion of Polygonatum sibiricum is 350 parts, 380 parts, 400 parts, 425 parts, 450 parts, 480 parts or 500 parts; the mass portion of Nardostachys chinensis is 150 parts, 170 parts, 190 parts, 210 parts or 230 parts; the mass portion of Poria cocos is 100 parts, 125 parts, 150 parts, 175 parts or 200 parts.

[0054] The following are several groups of examples to conduct model test studies on the Chinese medicine composition for treating cardiac blood stasis type arrhythmia, the preparation method and the effect of treating cardiac blood stasis type arrhythmia provided by the present invention.

[0055] Example 1

[0056] The Chinese medicine composition for treating arrhythmia of cardiac blood stasis type provided in this embodiment is composed of the following raw materials in parts by weight: 300 parts of Codonopsis pilosula, 60 parts of Panax notoginseng, 400 parts of Polygonatum sibiricum, 200 parts of Nardostachys grandiflora and 160 parts of Poria cocos.

[0057] In this embodiment, the Chinese medicine composition is prepared according to the following steps:

[0058] S1. Prepare raw materials of Codonopsis pilosula, Panax notoginseng, Polygonatum sibiricum, Nardostachys grandiflora and Poria cocos according to mass proportions; specifically, weigh each raw material according to 1 g per proportion.

[0059] S2, extracting volatile oil from Nardostachys grandiflora by water extraction;

[0060] Weigh 200 g of Nardostachys grandiflora, add 10 times the amount of water to extract the volatile oil of Nardostachys grandiflora, and collect the extracted aqueous solution in another container.

[0061] S3. The Panax notoginseng is extracted by ethanol reflux method to obtain Panax notoginseng filtrate and Panax notoginseng residues. The Panax notoginseng filtrate is concentrated under reduced pressure to obtain Panax notoginseng paste. The Panax notoginseng residues are decocted with water to obtain Panax notoginseng decoction.

[0062] Weigh 60 g of Panax notoginseng powder, extract it with 10 times the amount of 80% ethanol under reflux twice, each time for 2 hours, and filter; combine the two filtrates and concentrate under reduced pressure to obtain Panax notoginseng paste; add 10 times the amount of water to the remaining Panax notoginseng residue and boil it twice, the first time for 2 hours and the second time for 1.5 hours, and combine the two decoctions to obtain Panax notoginseng decoction liquid.

[0063] S4, after mixing Codonopsis pilosula and Polygonatum sibiricum, a decoction is obtained by a decoction method; then filtering and concentrating to obtain a Codonopsis pilosula and Polygonatum sibiricum decoction; and combining it with the Panax notoginseng decoction in step S3, a mixed filtrate is obtained by a water extraction and alcohol precipitation method.

[0064] Weigh 300 g of Codonopsis pilosula and 400 g of Polygonatum sibiricum, add 10 times the amount of water and decoct twice, the first time for 2 hours and the second time for 1.5 hours, combine the two filtrates to obtain a Codonopsis pilosula and Polygonatum sibiricum decoction; mix the Codonopsis pilosula and Polygonatum sibiricum decoction and the Panax notoginseng decoction evenly, filter, concentrate the obtained filtrate to a relative density of 1.20-1.30 (60°C), add ethanol to make the alcohol content reach 65%, stir, let stand for 24 hours, filter, and obtain a mixed filtrate.

[0065] S5. Extracting Poria cocos by ethanol reflux method to obtain Poria cocos extract, combining the extract with the mixed filtrate of step S4, and concentrating under reduced pressure to obtain a thick paste.

[0066] Weigh 160 g of Poria cocos, soak for 12 hours, add 9 times the amount of water, heat in a water bath and reflux for extraction twice, the first time for 7.5 hours and the second time for 1.5 hours, and combine the two filtrates to obtain a Poria cocos extract; after combining the Poria cocos extract with the mixed filtrate of step S4, concentrate under reduced pressure to obtain a thick paste.

[0067] S6, mixing the Panax notoginseng paste of step S3 and the thick paste of step S5; then adding the nardostachys volatile oil of step S2, mixing evenly, and freeze-vacuum drying to obtain a Chinese medicine composition.

[0068] Example 2 Pharmacokinetic Study of Composition

[0069] The purpose of this example is to use the Chinese medicine composition provided in Example 1, take the positive control drug Wenxin granules as a reference, and study the pharmacokinetics of the Chinese medicine composition by designing the dosage of the Chinese medicine composition and conducting a model test.

[0070] 2.1. Determination of drug concentration for intragastric administration

[0071] The clinical dosage of Wenxin Granule is 15g / d. The equivalent dose ratio of rats to humans is calculated as 6.3, and the rat dosage is 15g / 70kg×6.3=1.35g / kg. The content of main saponins (notoginseng saponin R1, ginsenoside Rg1 and ginsenoside Rb1) in Wenxin Granule is 5.74858mg / g, and the main saponins in the composition are 13.33279mg / g. Therefore, the dosage of the composition is

[0072] 1.35 g / kg x 5.74858 mg / g / 13.33279 mg / g = 0.58 g / kg = 58 mg / 100 g, i.e. 1 mL is administered per 100 g of animal, so the formulated composition is 58 mg / mL.

[0073] 2.2 Preparation of drug solution for intragastric administration

[0074] Accurately weigh an appropriate amount of the powder of the Chinese medicine composition in Example 1 and dissolve it in physiological saline to prepare a solution with a concentration of 58 mg / mL.

[0075] 2.3 Plasma sample processing

[0076] Take 200 μL of rat plasma sample, add 10 μL of internal standard and 600 μL of methanol, vortex for 1 min, and rotate at 13300 r·min. -1 After centrifugation for 15 min, the supernatant was aspirated and dried in a low-temperature vacuum concentrator. The residue was re-dissolved in 100 μL of methanol and ultrasonicated for 10 min at 13300 r / min. -1 After centrifugation for 15 min, the supernatant was filtered through a 0.22 μm microporous membrane and 2 μL was aspirated for LC-MS analysis.

[0077] 2.4. Pharmacokinetic study of the main components of the composition

[0078] Male SD rats (230±20g) were randomly divided into 2 groups, 6 in each group, namely, a blank group and a combination group. The rats were fasted for 12h before administration and were allowed to drink water freely. Blood was collected from the abdominal aorta of the blank group, and 0.4mL of blood was collected from the fundus venous plexus of the drug-treated group (composition 58mg / 100g) at 0.083h, 0.167h, 0.25h, 0.5h, 0.75h, 1h, 1.5h, 2h, 3h, 4h, 6h, 8h, and 12h after administration in a sodium heparin tube, centrifuged at 4°C and 13300r / min for 15min, and the upper plasma sample was separated and stored at -80°C. Pharmacokinetic parameters (C max , T max , T 1 / 2 , AUC, MRT, etc.) and plot the blood drug concentration-time curve. The blood drug concentration-time curve of the composition is as follows Figure 1 As shown, the pharmacokinetic parameters are shown in Table 1; Figure 1Among them, A is ginsenoside Rg1, B is ginsenoside Rb1, C is protopanaxatriol, and D is 7-OH coumarin.

[0079] Table 1 Pharmacokinetic parameters of the four main components of the composition in rat plasma ( n=6)

[0080] Pharmacokinetic parameters Ginsenoside Rg1 Ginsenoside Rb1 Protopanaxatriol 7-OH Coumarin <![CDATA[C max (μg / L)]]> 8.64±2.51 39.51±12.16 593.31±93.58 47.60±11.14 <![CDATA[AUC (0-t) (μg / L·h)]]> 36.06±10.10 302.31±98.84 3724.30±895.20 352.61±87.74 <![CDATA[T 1 / 2 (h)]]> 5.85±2.32 15.43±9.67 15.47±7.47 17.45±12.90 <![CDATA[T max (h)]]> 0.63±0.21 5.67±1.97 0.08±0.00 2.08±0.49 <![CDATA[MRT (0-t) (h)]]> 4.12±0.95 6.50±0.50 5.52±0.94 5.43±0.20

[0081] Example 3 Evaluation of the therapeutic effect of Chinese medicine composition on arrhythmia of blood stasis type

[0082] To further verify whether the composition has a therapeutic effect on cardiac blood stasis type arrhythmias, a rat model of myocardial ischemia-reperfusion injury was established, a pharmacodynamic evaluation was performed, and a multi-omics joint analysis was combined to study the mechanism of action, and finally the relevant pathways were verified.

[0083] 3.1. Trial Drugs

[0084] Composition: The Chinese medicine composition prepared in Example 1 was used. The positive control was diltiazem, a Western medicine; the positive control was Wenxin granule, a Chinese medicine.

[0085] 3.2 Preparation of drug solution for intragastric administration

[0086] Preparation of positive control western medicine diltiazem (DZ) solution: After the diltiazem tablets are crushed, an appropriate amount is accurately weighed and dissolved in 5% sodium carboxymethylcellulose suspending agent to prepare a suspension with a concentration of 12.5 mg / mL.

[0087] Chinese medicine positive control Wenxin granules: Accurately weigh an appropriate amount of Wenxin granules powder and dissolve it in physiological saline to prepare a solution with a concentration of 58 mg / mL.

[0088] Preparation of the Chinese medicine composition for intragastric administration: Accurately weigh an appropriate amount of the composition powder and dissolve it in physiological saline to prepare a solution with a concentration of 58 mg / mL.

[0089] 3.3 Animal model preparation

[0090] Male SD rats were randomly divided into sham operation group (blank group Control), model group (Model), western medicine positive group (western medicine diltiazem (DZ)), Chinese medicine positive group (Wenxin granule group (WXKL)), and Chinese medicine combination group (ZHW) after 7 days of adaptive growth.

[0091] The drug was administered continuously for 2 weeks, once a day in the morning. On the 15th day, the rat myocardial ischemia-reperfusion model was prepared, and the sham operation group was not subjected to ischemia-reperfusion treatment. The rats were fasted for 12 hours before modeling, and the drug was administered once. After 15 days of intragastric administration, the rats were anesthetized by intraperitoneal injection of 10% ulanose (10mL / kg), the hair on the neck and chest was removed, and the parameters of the small animal ventilator were adjusted to tidal volume: 6.2mL, respiratory time ratio of 1:1, and respiratory rate of 82 times / minute. After the rats were unresponsive to anesthesia, their limbs and head were fixed in a supine position, the neck and chest were disinfected, the trachea was intubated for ventilation, and the electrocardiogram was connected, followed by thoracotomy. The left side of the sternum was incised, the skin and muscles were separated, the heart was exposed between the 3rd and 4th intercostal spaces, the pericardium was separated, the position of the left anterior descending coronary artery was determined, and a 6-0 suture with a needle was inserted about 2 mm below the lower edge of the left atrial appendage, and the needle was removed from the left side of the venous plexus at the junction of the pulmonary artery cone and the left atrial appendage for slipknot ligation. The ventricular muscle surface and apex at the distal end of the ligation were observed to turn white, and the electrocardiogram showed a significant elevation of the ST segment, indicating successful ischemia. After ligation for 30 minutes, the line was loosened for reperfusion for 3 hours, and the muscles and skin were sutured with 4-0 sutures and 2-0 sutures, respectively. After the rats recovered spontaneous breathing, the endotracheal tube was removed and placed on an electric blanket. The sham operation group only threaded the line without ligation, and the rest of the operations were the same as the model group.

[0092] The electrocardiogram during the modeling period was Figure 2 As shown, A is before ligation, B is during ischemia, and C is during reperfusion. During the ligation period, the ST segment of the rat ECG was significantly elevated, and during the reperfusion period, the ST segment gradually decreased, indicating that the rat myocardial ischemia-reperfusion model was successfully established.

[0093] 3.4 Experimental sample collection

[0094] After the perfusion, blood was collected from the abdominal aorta for serum biochemistry and Elisa tests, the heart tissue was separated and washed with pre-cooled saline perfusion, three rat heart tissues were stored at -20°C for TTC staining, three were placed in 4% paraformaldehyde tissue fixative for HE, Masson, and Tunel staining, and the rest were quickly placed on dry ice and transferred to -80°C for storage for metabolomics, transcriptomics, Western blot, and PCR analysis.

[0095] 3.5 Test results

[0096] 3.5.1 TTC staining

[0097] The rat hearts of each group were placed in a -20°C refrigerator for 30 minutes, cut into 5 pieces starting from the apex of the heart, placed in a 2% TTC solution, reacted at 37°C in the dark for 20 minutes, and flipped once every 5 minutes to ensure uniform staining. The hearts were taken out and fixed in paraformaldehyde for 4 hours, then photographed. The images were analyzed using image analysis software and the myocardial infarction area was calculated. Figure 3 As shown, where A is the TTC staining image of the I / R model, and B is the percentage of myocardial infarction area ( n=3); Note: **p<0.01 compared with the sham operation group.

[0098] See also Figure 3 Compared with the sham operation group (Sham), the myocardial infarction area of ​​the model group (Model) rats was significantly increased to (31.94±3.46)% (p<0.01), which combined with the electrocardiogram results further indicated that the myocardial ischemia-reperfusion model was successfully established.

[0099] 3.5.2. Serum biochemical index determination

[0100] After blood was collected from the abdominal aorta of rats, the blood was left to stand for 2 hours, centrifuged at 4000r / min for 15 minutes, and the supernatant was collected and packaged. The aspartate aminotransferase (AST), serum α-hydroxybutyrate dehydrogenase (α-HBDH), creatine kinase (CK), creatine kinase isoenzyme (CK-MB) and isocitrate dehydrogenase (LDH) were measured using an automatic biochemical analyzer. Figure 4 As shown, where: ( n≥6) A is AST, B is CK-MB, C is CK, D is α-HBDH, and E is LDH; Note: Compared with the blank group *p<0.05, compared with the model group # p<0.05.

[0101] See also Figure 4 Compared with the blank group, the AST, CK, CK-MB, α-HBDH and LDH in the model group were significantly increased (p < 0.05). After drug administration, the AST, CK, CK-MB, α-HBDH and LDH in the model group were significantly improved (p < 0.05).

[0102] 3.5.3. Serum cTNI, cTnT, and NT-proBNP levels

[0103] According to the kit instructions, the ELISA method was used to detect the levels of cardiac troponin I (cTNI), pro-B-type natriuretic peptide (NT-proBNP) and cardiac troponin T (cTnT) in the serum of each group. Figure 5 As shown, where A is NT-proBNP, B is TnT, and C is cTnI ( n≥6); Note: Compared with the blank group*p<0.05, compared with the model group # p<0.05.

[0104] See also Figure 5 Myocardial ischemia-reperfusion injury significantly increased the expression levels of NT-proBNP, cTnT, and cTnI in serum (P < 0.05). After administration of the drug, the expression levels of NT-proBNP, cTnT, and cTnI significantly decreased (p < 0.05).

[0105] 3.5.4 HE staining

[0106] The fixed heart tissue was dehydrated by an automatic dehydrator, embedded, and sliced ​​in paraffin. The slices were dewaxed to water, stained with hematoxylin for 10-20 minutes, dehydrated with 85% gradient alcohol for 5 minutes, stained with eosin for 5 minutes, dehydrated and sealed, and finally the slices were imaged using a digital slice scanner. Figure 6 As shown, n=3, A is the blank group, B is the model group, C is the positive drug group of western medicine, D is the positive drug group of traditional Chinese medicine, and E is the combination group. Note: Myocardial fiber degeneration and necrosis (green arrow), neutrophil (red arrow), hemorrhage (blue arrow)

[0107] See also Figure 6 The myocardial cells of the rats in the sham operation group were round, arranged regularly, and had clear boundaries. The myocardial cells of the rats in the model group were edematous, with blurred boundaries and disordered arrangement. Some myocardial fibers showed blurred cell structure, cytoplasm dissolution, nuclear condensation or fragmentation. Some myocardial fibers were atrophied and broken, with dark cytoplasm and nuclear condensation. Degeneration and necrosis of epicardium and myocardial fibers were observed, accompanied by inflammatory cell infiltration. After administration, compared with the model group, the bleeding of myocardial cells, degeneration and necrosis of myocardial fibers, and infiltration of inflammatory cells in the drug administration group were reduced.

[0108] 3.5.5 Masson staining

[0109] The fixed heart tissue was dehydrated in an automatic dehydrator, embedded, and sliced ​​in paraffin. The slices were dewaxed to water, incubated with potassium dichromate overnight, differentiated and blued with hematoxylin, stained with Ponceau fuchsin solution for 10 minutes, treated with phosphomolybdic acid solution for 1 minute, stained with aniline blue for about 2 minutes, dehydrated with gradient alcohol, and sealed. Finally, the slices were imaged with a digital slice scanner. The image analysis system was used to determine the positive expression area in the collected images, and the percentage of positive expression area was calculated = positive expression area / field of view area (pixel area). The results are shown in Figure 7 As shown. Among them: n = 3; A is the blank group, B is the model group, C is the positive drug group of western medicine, D is the positive drug group of traditional Chinese medicine, E is the combination group, and F is the fibrosis area; Note: Compared with the blank group *p<0.05, compared with the model group# p<0.05.

[0110] See also Figure 7 The results showed that compared with the sham operation group, the myocardial interstitial collagen fibers in the model group increased significantly to (10.04±1.07)%. After drug administration, the collagen fibers in the drug administration group were significantly reduced to varying degrees compared with the model group.

[0111] 3.5.6 Tunel staining

[0112] The fixed heart tissue was dehydrated by an automatic dehydrator, embedded, paraffin sectioned, stained, sealed, and finally examined under a microscope. The normal cells and apoptotic cells in the image were counted manually, and the percentage of apoptotic cells was calculated as apoptotic cells / (normal cells+apoptotic cells×100%). The results are shown in Figure 2. Figure 8 and Fig. 9 As shown. Wherein: n = 3; Figure 8 Tunel staining diagram of rats in each group. Fig. 9 is the apoptosis rate of myocardial cells in each group of rats; Note: compared with the blank group *p<0.05, compared with the model group # p<0.05.

[0113] See also Figure 8 and Fig. 9 Compared with the sham operation group, the green fluorescence of cardiomyocytes in the model group was significantly increased, and the apoptosis rate of cardiomyocytes was significantly increased to (25.48±7.07)%. Compared with the model group, the apoptosis rate of cardiomyocytes in the combination administration group was significantly reduced.

[0114] Example 4 Metabolomics Study

[0115] In this example, the Chinese medicine composition provided in Example 1 was used to study the metabolic mechanism of the Chinese medicine composition through a model experiment.

[0116] 4.1 LC-MS conditions

[0117] Chromatographic conditions: ultra high performance liquid phase C18 chromatographic column (2.1 mm × 100 mm, 1.7 μm), column temperature: 30 ° C; flow rate: 0.3 mL / min; injection volume: 2 μL; mobile phase: 0.028% ammonium acetate-0.05% formic acid aqueous solution (A): acetonitrile (B). Elution program: 0-1.5 min, 1% acetonitrile; 1.5-23 min, 1%-60% acetonitrile; 23-24 min, 60%-98% acetonitrile; 24-27 min, 98% acetonitrile; 27-27.1 min, 98%-1% acetonitrile, 27.1-33 min, 1% acetonitrile.

[0118] Mass spectrometry conditions: electrospray ion source, positive / negative ion mode detection; capillary voltage: +5000 / -4500 V; fragmentation voltage: 40 V; curtain gas: 35 psi; nebulization temperature: 550°C; parent ion mass scanning range: m / z 70-1200 Da; nebulizer: 50 psi, auxiliary gas: 50 psi.

[0119] 4.2 Preparation of Cardiac Metabolic Samples

[0120] After the heart sample was thawed in a 4°C refrigerator, 50 mg of the heart sample was accurately weighed, 600 μL of pre-cooled 80% methanol-water solution was added, pre-cooled steel beads were added, homogenized for 2 min, ultrasonicated for 10 min, allowed to stand at -20°C for 20 min, centrifuged at 12000 r / min and 4°C for 15 min, 500 μL of the supernatant was concentrated to dryness in a low-temperature vacuum concentrator, 200 μL of 50% methanol was added for re-dissolution, vortexed for 1 min, ultrasonicated for 10 min, centrifuged at 12000 r / min and 4°C for 15 min, and the supernatant was taken for injection analysis. At the same time, 2 μL of the mixture was taken from each sample as a quality control (QC) sample.

[0121] 4.3 Data Analysis

[0122] The mass spectrometry data acquisition software was used to acquire data, and the metabolites were analyzed and identified in combination with the data analysis software, data quantification software, metabolome database, and online HMDB metabolite chemical database (https: / / hmdb.ca / ).

[0123] The original data were analyzed by principal component analysis (PCA) to compare the control group and the model group. The volcano diagram of differential protein expression was visualized with |logFC|>0.5 and p-value<0.05 as the screening conditions, and the metabolic profiles of differential metabolites in the hearts of rats in each group were analyzed by cluster analysis.

[0124] 4.4 Metabolic profile analysis of model group and blank group

[0125] Auto scaling was performed on the blank group, model group and quality control sample data in positive and negative ion modes, and PLS analysis was performed on their metabolic profiles, such as Fig.10 As shown in Figure 1. Among them: A is the negative ion mode, and B is the positive ion mode.

[0126] See also Fig.10 In the positive and negative ion modes, the QC samples were well clustered together, indicating that the instrument had good stability. In addition, there was an obvious intergroup separation phenomenon between the blank group and the model group, indicating that there were obvious differential metabolites in the heart tissue of the I / R model group compared with the blank group.

[0127] 4.5. Screening of differential metabolites

[0128] The volcano map of differential protein expression was visualized using |logFC|>0.28 and p-value<0.05 as the screening conditions. Fig.11 , Fig.12 , Fig.13 and Fig.14 shown. Fig.11 The volcano plots of cardiac tissue metabolites in the blank group and the model group, where: A is the negative ion mode; B is the positive ion mode; Fig.12 , Fig.13 and Fig.14 This is the volcano plot of cardiac tissue metabolites in the drug-treated group and the model group, where: Fig.12 Positive Western medicine group vs model group, Fig.13 Positive Chinese medicine group vs model group, Fig.14 For the combination group vs the model group, Fig.12 , Fig.13 and Fig.14 The meaning of the vertical and horizontal coordinates in Fig.11 .

[0129] See also Fig.11 (A) In the negative ion mode, the I / R model group upregulated 36 metabolites and downregulated 11 metabolites compared with the blank group; see Fig.11 (B) In the positive ion mode, the I / R model group upregulated 27 metabolites and downregulated 12 metabolites compared with the blank group.

[0130] See also Fig.12 , Fig.13 and Fig.14 In the negative ion mode, the western medicine positive group upregulated 7 metabolites and downregulated 9 metabolites, and in the positive ion mode, a total of 18 metabolites were upregulated and 4 metabolites were downregulated ( Fig.12 ); Chinese medicine positive drugs up-regulated 28 metabolites and down-regulated 9 metabolites in negative ion mode, down-regulated 11 metabolites and up-regulated 21 metabolites in positive ion mode ( Fig.13 ); the combination upregulated 22 metabolites and downregulated 12 metabolites in negative ion mode, and upregulated 19 metabolites and downregulated 10 metabolites in positive ion mode ( Fig.14 ).

[0131] 4.6 Metabolic pathway enrichment analysis

[0132] The differential metabolites between the model group and the blank group, as well as the differential metabolites between each drug group and the model group were subjected to metabolic pathway enrichment analysis. Fig.15 As shown, A is the model group vs. blank group, B is the western medicine positive group vs. model group, C is the traditional Chinese medicine positive group vs. model group, and D is the combination group vs. model group.

[0133] See also Fig.15 , the differential metabolites between the model group and the blank group participated in the regulation of 37 metabolic pathways. After administration, the composition could regulate 23 metabolic pathways in the model group. It mainly participated in the regulation of aminoacyl-tRNA biosynthesis, valine, leucine and isoleucine biosynthesis, glycine, serine and threonine metabolism, amino sugar and nucleotide sugar metabolism, pyrimidine metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, alanine, aspartate and glutamate metabolism, cysteine ​​and methionine metabolism, phenylalanine metabolism, pentose and glucuronic acid interconversion, fructose and mannose metabolism, sphingosine metabolism, propionate metabolism, galactose metabolism, glutathione metabolism, steroid hormone biosynthesis, unsaturated fatty acid biosynthesis 17 metabolic pathways.

[0134] Example 5 Study on the Mechanism of Action for the Treatment of Arrhythmias with Blood Stasis

[0135] In this example, the Chinese medicine composition provided in Example 1 was used to study the mechanism of action of the Chinese medicine composition in treating arrhythmias of the blood stasis type through a model experiment.

[0136] 5.1 Transcriptomic Analysis

[0137] 5.1.1. RNA extraction

[0138] Weigh an appropriate amount of heart tissue in the sample collection of 3.4 in Example 3, add 1.5mL Trizol reagent and 375μL chloroform, vortex for 15s, stand at room temperature for 2-3min, centrifuge at 2000r / min and 4℃ for 15 minutes, draw the upper aqueous phase into a new centrifuge tube, add an equal volume of isopropanol, mix well and stand at room temperature for 10min. Centrifuge at 12000r / min and 4℃ for 10min, discard the supernatant, add 1mL 75% ethanol prepared with DEPC water to wash the precipitate, centrifuge at 12000r / min and 4℃ for 3min, discard the supernatant, stand at room temperature for 2-3min, dry, add 100μL DEPC water, and after complete dissolution, take a small amount of sample RNA integrity and DNA contamination by agarose gel electrophoresis, use ultra-micro UV-visible photometer to detect RNA purity, and use bioanalyzer to detect RNA integrity, and the rest is stored at -80℃.

[0139] 5.1.2 Library construction and quality control

[0140] The library was constructed according to the instructions of the library construction kit. After the construction was completed, a nucleic acid protein quantifier was used for preliminary quantification, and the library was diluted to 1.5 ng / μL. The insert size of the library was then detected using a bioanalyzer. After the insert size met expectations, qRT-PCR was used to accurately quantify the effective concentration of the library.

[0141] 5.1.3 Sequencing

[0142] After the library was qualified, it was sequenced on the sequencing platform and 150bp paired-end reads were generated.

[0143] 5.1.4 Data Analysis

[0144] The image data of the sequencing fragments measured by the high-throughput sequencer were converted into sequence data by CASAVA base recognition, and the sequence data with adapters, sequence data containing N (indicating that the base information could not be determined), and low-quality sequence data (sequence data with base numbers of Qphred≤20 accounting for more than 50% of the total data length) were filtered out. At the same time, Q20, Q30 and GC content were calculated for the remaining quality control data. The RNA sequence alignment software was used to construct the index of the reference genome, and the paired-end quality control data were aligned with the reference genome. The gene expression difference analysis software was used to analyze the differentially expressed genes between the two groups, and the Benjamini&Hochberg method was used to adjust the P value. The corrected P value |log2FC| was used as the threshold for significant differential expression. GO enrichment analysis and KEGG enrichment analysis of differentially expressed genes were implemented using R software.

[0145] 5.2 Screening of differentially expressed genes in the transcriptome

[0146] To further screen the differentially expressed genes of the combination for I / R intervention, a volcano plot of differentially expressed genes was drawn, such as Fig.16 , Fig.17 and Fig.18 As shown, where: Fig.16 For the blank group and the model group, Fig.17 The Chinese medicine positive group and the model group, Fig.18 There are combination group and model group.

[0147] See also Fig.16 , Fig.17 and Fig.18 Compared with the blank group, the model group upregulated 2145 genes and downregulated 1929 genes. After drug administration, compared with the model group, the combination group upregulated 572 genes and downregulated 335 genes. By taking the intersection of the differentially expressed genes between the blank group and the model group and the differentially expressed genes between the drug administration group and the model, it was found that the combination group could upregulate 196 differentially expressed genes in the model group and downregulate 105 differentially expressed genes in the model group.

[0148] 5.3. Annotation of GO biological functions of differentially expressed genes in the transcriptome

[0149] GO enrichment analysis was performed on the differentially expressed genes in rats with I / R model treated with the combination. The results are as follows: Fig.19 shown.

[0150] See also Fig.19 When the combination intervenes in the I / R model, the biological processes (BP) involved include the regulation of the metabolic process of nucleobase-containing compounds, the metabolic process of xenobiotics, the cellular metabolic process, the regulation of nucleic acid template transcription, the cell response to exogenous stimuli, etc. The cell components (CC) involved mainly include membrane-bound organelles, intracellular membrane-bound organelles, intercellular connections, intracellular parts, etc. The molecular functions (MF) involved mainly include metal ion binding, cation binding, transition metal ion binding, ion binding, etc.

[0151] 5.4 KEGG enrichment analysis of differentially expressed genes in transcriptomes

[0152] KEGG enrichment analysis was performed on the differentially expressed genes in rats with I / R model treated with the combination. The results are as follows Fig. 20 As shown, the pathways in which it is mainly involved include the JAK-STAT signaling pathway, drug metabolism-cytochrome P450, inflammatory mediator regulation of TRP channels, niacin and nicotinamide metabolism, etc.

[0153] 5.5 RT-PCR analysis

[0154] About 20 mg of heart tissue frozen at -80°C was weighed, RNA was extracted according to the instructions of the rapid RNA extraction kit, and the concentration and purity of total RNA were determined using an ultra-micro UV spectrophotometer. Total RNA was reverse transcribed into cDNA according to the reaction system of the reverse transcription kit, diluted to a cDNA concentration of 400 ng / μL, and then RT-PCR amplification was performed according to the instructions of the kit (Table 2). The relative expression levels of the genes to be tested were calculated. The primer sequences of the genes to be tested were designed by Primer-BLAST, and the information is shown in Table 3.

[0155] Table 2 RT-qPCR reaction system of differentially expressed genes

[0156]

[0157] The primer sequences are shown in Table 3.

[0158] Table 3 Primer sequences

[0159]

[0160]

[0161] The genes related to the JAK2-STAT3 signaling pathway were detected by RT-PCR. Fig.21 As shown, A is JAK2, B is STAT3, C is BCL2, and D is MCL1; Note: **p<0.05 compared with the blank group, #p<0.05, ##p<0.01 compared with the model group.

[0162] See also Fig.21 Compared with the blank group, I / R caused a significant decrease in the gene expression of JAK2 and MCL1, and a significant increase in the gene expression of STAT3 and BCL2 (p < 0.05). After administration, JAK2 and MCL1 were significantly upregulated, and STAT3 and BCL2 were significantly downregulated (p < 0.05).

[0163] 5.6 Western Blot Detection

[0164] Weigh 80 mg of heart tissue from each group, wash twice with saline, add 800 μL of RIPA lysis buffer containing 1% PMSF for homogenization and lysis, stand on ice for 30 minutes after complete homogenization, centrifuge and take the supernatant as the total protein, and perform protein denaturation after the protein concentration is determined according to the BCA protein concentration determination kit. Then perform SDS-PAGE gel electrophoresis separation, transfer to the membrane, block with blocking solution for 15 minutes, transfer to the primary antibody solution and incubate overnight at 4°C. The dilution ratio of the antibody is shown in Table 4. After rinsing with washing solution 3 times, incubate with the secondary antibody for 1.5 hours, wash again 3 times, perform ECL luminescence development, and finally use Image Lab software for optical density measurement and analysis. The dilution ratio of the antibody is shown in Table 4.

[0165] Table 4 Antibody dilution ratio

[0166] Antibody Dilution ratio β-actin 1：1500 JAK2 1：1500 P-JAK2 1：800 STAT3 1：2000 P-STAT3 1：5000 MCL1 1：800 BCL2 1：5000 HPR-IgG goat anti-rabbit 1：8000

[0167] The results are as follows Fig. 22 and Fig.23 As shown, Fig. 22 Middle: A is the expression of JAK2 protein, B is the expression of STAT3 protein, C is the expression of BCL2 protein, and D is the expression of MCL1 protein; Fig.23 Western Blot band diagram; Note: Compared with the blank group, **p<0.01, *p0.05, compared with the model group, ##p<0.01, #p<0.05.

[0168] See also Fig. 22 and Fig.23Compared with the blank group, I / R caused a significant decrease in the protein expression of p-JAK2, p-STAT3, MCL1, and BCL2 (p < 0.05). After administration, the composition had a significant upregulation effect on p-JAK2, p-STAT3, MCL1, and BCL2 (p < 0.05).

[0169] 5.7 Analysis of JAK2-STAT3 signaling pathway

[0170] The above studies found that the intervention mechanism of the composition on I / R model rats is mainly related to the JAK-STAT signaling pathway, drug metabolism-cytochrome P450 and TRP channel inflammatory mediator regulation. The JAK-STAT signaling pathway is involved in the expression of cytokines, growth factors and hormones, and has been proven to play an important role in the protection of ischemia-reperfusion injury. The JAK-STAT signaling pathway is involved in cell growth, development, proliferation, apoptosis and necrosis. After myocardial ischemia-reperfusion, the expression of JAK2, STAT3 and BCL2 proteins was significantly reduced, while the apoptosis rate of myocardial cells was increased. Among them, STAT3 can be activated by multiple factors, dimerize, enter the cell nucleus to activate transcription, thereby causing changes in the cell cycle. In addition, SAT3 can significantly reduce the level of inflammatory factors, protect myocardial cells by inhibiting inflammatory responses and increasing cell activity. The composition can improve myocardial tissue damage by upregulating the downregulation of BCL2 caused by myocardial ischemia-reperfusion, indicating that the composition can inhibit the apoptosis of myocardial cells by activating the JAK2-STAT3 signaling pathway, and has a therapeutic effect on cardiac blood stasis type arrhythmias ( Fig.24 ).

[0171] The specific implementation methods described above further illustrate the objectives, technical solutions and beneficial effects of the present invention in detail. It should be understood that the above description is only a specific implementation method of the present invention and is not intended to limit the scope of protection of the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A Chinese medicine composition for treating cardiac blood stasis type arrhythmia, characterized in that: The Chinese medicine composition is composed of the following raw materials in parts by weight: 250-370 parts of codonopsis pilosula, 55-100 parts of notoginseng, 350-500 parts of polygonatum, 150-230 parts of nardostachys grandiflora and 100-200 parts of tuckahoe.

2. The Chinese medicine composition for treating cardiac blood stasis type arrhythmia according to claim 1, characterized in that: The Chinese medicine composition consists of the following raw materials in parts by weight: 300 parts of Codonopsis pilosula, 60 parts of Panax notoginseng, 400 parts of Polygonatum sibiricum, 200 parts of Nardostachys grandiflora and 160 parts of Poria cocos.

3. The Chinese medicine composition for treating cardiac blood stasis type arrhythmia according to claim 2, characterized in that: The Chinese medicine composition has a significant up-regulating effect on the expression of JAK2 protein and MCL1 protein, and a significant down-regulating effect on the expression of STAT3 protein and BCL2 protein.

4. The method for preparing the Chinese medicine composition for treating cardiac blood stasis type arrhythmia according to claim 1, characterized in that: The preparation method comprises the following steps: S1. Prepare raw materials of Codonopsis pilosula, Panax notoginseng, Polygonatum sibiricum, Nardostachys grandiflora and Poria cocos according to weight; S2, extracting volatile oil from Nardostachys grandiflora by water extraction; S3, extracting Panax notoginseng by ethanol reflux method to obtain Panax notoginseng filtrate and Panax notoginseng residues, concentrating the Panax notoginseng filtrate under reduced pressure to obtain Panax notoginseng paste, and decocting the Panax notoginseng residues with water to obtain Panax notoginseng decoction; S4, after mixing Codonopsis pilosula and Polygonatum sibiricum, a decoction is obtained by a decoction method; then filtering and concentrating to obtain a Codonopsis pilosula and Polygonatum sibiricum decoction; and combining the Panax notoginseng decoction in step S3, a mixed filtrate is obtained by a water extraction and alcohol precipitation method; S5, extracting Poria cocos by ethanol reflux method to obtain Poria cocos extract, combining it with the mixed filtrate of step S4, and concentrating under reduced pressure to obtain a thick paste; S6, mixing the notoginseng paste of step S3 and the thick paste of step S5; then adding the nardostachys volatile oil of step S2, mixing evenly, and freeze-vacuum drying to obtain a Chinese medicine composition.

5. The preparation method according to claim 4, characterized in that: In the step S2, 10 times the mass of water is added to the nardostachys grandiflora for extraction to obtain nardostachys grandiflora volatile oil.

6. The preparation method according to claim 4, characterized in that: The specific process of step S3 is: Reflux extraction was performed twice with 10 times the mass of ethanol, each time for 2 hours, and filtered; the two filtrates were combined and concentrated under reduced pressure to obtain Panax notoginseng paste; Add 10 times the amount of water to the Panax notoginseng residue and boil it twice, the first time for 2 hours and the second time for 1.5 hours. The decoctions are combined to obtain the Panax notoginseng decoction.

7. The preparation method according to claim 4, characterized in that: The specific process of step S4 is: Mix Codonopsis pilosula and Polygonatum sibiricum, add 10 times the weight of water and boil twice, the first time for 2 hours, the second time for 1.5 hours, and obtain the Codonopsis pilosula and Polygonatum sibiricum decoction and combine it with the above decoction; The decoctions of Codonopsis pilosula and Polygonatum sibiricum and Panax notoginseng are mixed evenly and filtered, and the obtained filtrate is concentrated to a relative density of 1.20-1.30, and ethanol is added to make the alcohol content reach 65%, stirred, allowed to stand for 24 hours, filtered, and a mixed filtrate is obtained.

8. The preparation method according to claim 4, characterized in that: The specific process of step S5 is: Soak the Poria cocos for 12 hours, add 9 times the weight of water, and heat under reflux in a water bath for 2 times, the first time for 7.5 hours and the second time for 1.5 hours, to obtain the Poria cocos extract; The Poria extract is combined with the mixed filtrate of step S4 and concentrated under reduced pressure to obtain a thick paste.

9. Use of the Chinese medicinal composition for treating cardiac blood stasis type arrhythmia as claimed in claim 1 or 2 in preparing a medicament for cardiac blood stasis type arrhythmia.

10. Use of the Chinese medicinal composition for treating arrhythmia of blood stasis type according to claim 1 or 2 in preparing a drug for arrhythmia of blood stasis type by activating JAK2-STAT3 signaling pathway to inhibit apoptosis of myocardial cells.

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