Preparation method of effective part of anchusa and application of effective part in prevention and treatment of myocardial injury

Abstract

The invention discloses a preparation method of an effective part of anchusa edulis, which comprises the following steps: (1) carrying out heating reflux extraction on anchusa edulis, and concentrating an extracting solution to obtain an anchusa edulis concentrated solution; and (2) adsorbing and eluting the concentrated solution of the anchusa through macroporous resin and / or polyamide resin to obtain the first effective part of the anchusa and / or the second effective part of the anchusa. The first effective part and / or the second effective part of the anchusa can recover human myocardial cell injury caused by verapamil, wherein the first effective part of the anchusa has the strongest activity in the aspects of promoting the overall activity of human myocardial cells, inhibiting the death of the human myocardial cells and promoting the proliferation of the human myocardial cells.

Description

Technical Field

[0001] The invention relates to the technical field of medicine, and in particular to a preparation method of an effective part of Herba Cynanchifoliae and an application thereof in preventing and treating myocardial damage. Background Art

[0002] Cardiovascular disease (CVD) is one of the major chronic diseases that endangers human health, and includes coronary heart disease, congenital heart disease, arrhythmias, and stable angina. The prevalence of CVD in my country is on a continuous rise, with an estimated 330 million people suffering from CVD. Conventional therapies for CVD in clinical practice mainly focus on antiplatelet aggregation, regulating lipid metabolism, improving myocardial blood supply, using cardioprotective agents, and regulating heart rate and blood pressure. However, complications such as bleeding, thrombosis, and heart failure can still occur. In contrast, traditional medicine, based on the "multi-component, multi-target, and multi-pathway" characteristics of traditional Chinese medicine, has demonstrated unique advantages in relieving clinical symptoms such as angina, chest tightness, shortness of breath, and palpitations. It can significantly reduce patient readmission rates and mortality, and has a potential positive effect on improving prognosis.

[0003] Bugloss is the dried aerial part of Anchusa italica Retz, a plant of the Boraginaceae family. It is primarily distributed in northern my country. Bugloss has the properties of removing dampness and heat, regulating abnormal black bile, refreshing the body and mind, strengthening the inner organs and senses, enhancing natural strength, and facilitating bowel movements. It is commonly used to treat insomnia, restlessness, headaches, slowed reaction times, and constipation.

[0004] There is no report in the prior art on the preparation method of the effective part of Herba Cynanchifoliae as claimed in the present invention and its use in preventing and treating myocardial damage. Summary of the Invention

[0005] Based on this, the present invention provides a method for preparing the effective part of Herba Cynanchifoliae, which comprises the following steps:

[0006] (1) extracting the herb by heating under reflux, and concentrating the extract to obtain a herb concentrate; and

[0007] (2) The Herba Glossinae concentrate is subjected to macroporous resin adsorption elution and / or polyamide resin adsorption elution to obtain the Herba Glossinae first effective fraction and / or the Herba Glossinae second effective fraction.

[0008] Furthermore, the bugloss is the dried aerial part of Anchusa italica Retz. of the Boraginaceae family.

[0009] Furthermore, the method of step (1) comprises the following steps: weighing an appropriate amount of Herba Cynanchii, adding ethanol to perform a first heating reflux extraction, filtering to obtain a first filtrate and a filter residue; adding ethanol to the filter residue to perform a second heating reflux extraction, filtering to obtain a second filtrate, combining the first filtrate and the second filtrate to obtain a Herba Cynanchii extract; and taking an appropriate amount of the Herba Cynanchii extract, performing reduced pressure rotary evaporation, and concentrating until there is no alcohol taste, thereby obtaining the Herba Cynanchii concentrate.

[0010] Furthermore, the method of step (1) does not include the step of defatting the Herba Lycopodii.

[0011] Furthermore, the volume percentage concentration of the ethanol is 60% to 80%, for example, about 70%.

[0012] Furthermore, in the first heating reflux extraction, the volume / mass (ml / g) ratio of the ethanol to the Herba Lycopodii Herba is 5-15, for example, about 10.

[0013] Furthermore, in the second heating reflux extraction, the volume / mass (ml / g) ratio of the ethanol to the Herba Lycopodii Herba is 4-8, for example, about 6.

[0014] Furthermore, the first heating reflux extraction time is 0.5 to 2 hours, for example, about 1 hour.

[0015] Furthermore, the second heating reflux extraction time is 0.5 to 2 hours, for example, about 1 hour.

[0016] Furthermore, the herbal drug concentration of the Herba Bovis is 0.1 to 0.5 g / ml, for example, about 0.29 g / ml.

[0017] Furthermore, the macroporous resin adsorption and elution method in step (2) comprises the following steps: loading the Herba Cynoglossi concentrated solution onto a macroporous resin for adsorption and allowing to stand; eluting according to the following procedure: (a) water, 1 to 3 column volumes, for example, about 2 column volumes, (b) 30% to 50% ethanol, for example, about 40% ethanol, 4 to 6 column volumes, for example, about 5 column volumes, (c) 60% to 80% ethanol, for example, about 70% ethanol, 6 to 10 column volumes, for example, about 8 column volumes, and (d) 90% to 100% ethanol, for example, about 95% ethanol, 4 to 6 column volumes, for example, about 5 column volumes; collecting the elution fractions of step (b) and / or step (c), concentrating under reduced pressure, and drying to obtain the Herba Cynoglossi first elution fraction and / or the Herba Cynoglossi first effective fraction.

[0018] Furthermore, the macroporous resin adsorption elution method further includes a macroporous resin pretreatment step.

[0019] Furthermore, the mass ratio between the crude drug amount in the Herba Bovis concentrate and the resin amount in the macroporous resin is about 1:(1-5), for example, about 1:about 2.5.

[0020] Furthermore, the macroporous resin is a HPD 300 macroporous resin column.

[0021] Furthermore, the volume ratio of the Herba Bolognese concentrated solution to the HPD 300 macroporous resin column is (5-9):(4-6), for example, about 7:about 5.

[0022] Furthermore, the diameter-to-height ratio of the HPD 300 macroporous resin column is about 1:7.

[0023] Furthermore, the flow rate of the sample loading is 0.1 to 0.5 BV / h, for example, about 0.28 BV / h.

[0024] Furthermore, the standing time is 8 to 16 hours, for example, about 12 hours.

[0025] Furthermore, the elution flow rate is 0.1 to 1 BV / h, for example, about 0.7 BV / h.

[0026] Furthermore, the first effective part of the Herba Bolognese is the part eluted with 60% to 80% ethanol, for example, about 70% ethanol.

[0027] Furthermore, the first elution portion of the Herba Bolognese is an elution portion with 30% to 50% ethanol, for example, about 40% ethanol.

[0028] Furthermore, the polyamide resin adsorption elution method in step (2) comprises the following steps: dissolving the first elution fraction of the Herba Bolognese with water, loading the fraction onto a polyamide resin for adsorption, and allowing the fraction to stand; eluting the fraction according to the following procedure: (A) water, 2 to 5 column volumes, for example, about 3.5 column volumes; (B) 50% to 70% ethanol, for example, about 60% ethanol, 5 to 10 column volumes, for example, about 7.5 column volumes; and (C) 90% to 100% ethanol, for example, about 95% ethanol, 3 to 7 column volumes, for example, about 5 column volumes; collecting the elution fraction with 50% to 70% ethanol, for example, about 60% ethanol, performing reduced pressure concentration, and drying to obtain the second effective fraction of the Herba Bolognese.

[0029] Furthermore, the polyamide resin adsorption and elution method further comprises a step of pretreating the polyamide resin.

[0030] Furthermore, the dissolution is ultrasound-assisted dissolution.

[0031] Furthermore, when water is added for dissolution, the volume / mass (L / g) ratio of the water to the first eluted portion of the Herba Bolognese is 0.01-0.1, for example, about 0.075.

[0032] Furthermore, the particle size of the polyamide resin is 30 to 60 meshes.

[0033] Furthermore, the ratio (g / L) of the mass of the first elution fraction of the Herba Bolognese to the column volume of the polyamide resin is (60-100):(1-5), for example, about 80:about 2.2.

[0034] Furthermore, the polyamide resin has an aspect ratio of about 1: about 5.5.

[0035] Furthermore, the flow rate of the sample loading is 0.1-1 BV / h, for example, about 0.65 BV / h.

[0036] Furthermore, the standing time is 8 to 16 hours, for example, about 12 hours.

[0037] Furthermore, the flow rate of the water elution is 0.5 to 1 BV / h, for example, about 0.82 BV / h.

[0038] Furthermore, the flow rate of the 50% to 70% ethanol, for example, about 60% ethanol elution, is 1 to 1.5 BV / h, for example, about 1.2 BV / h.

[0039] Furthermore, the flow rate of the 90% to 100% ethanol, for example, about 95% ethanol elution, is 1 to 3 BV / h, for example, about 1.9 BV / h.

[0040] According to another aspect of the present invention, there is provided an effective part of Herba Cynanchifoliae prepared by the above-mentioned preparation method;

[0041] The effective parts of the Herba Lycopersicum var. ameloides include the first effective part of the Herba Lycopersicum var. ameloides and / or the second effective part of the Herba Lycopersicum var. ameloides.

[0042] According to another aspect of the present invention, a Herba Cynoglossi pharmaceutical preparation is provided, which comprises the effective part of the Herba Cynoglossi and one or more pharmaceutically acceptable excipients.

[0043] Furthermore, the auxiliary material is selected from one or more of the following: diluent, wetting agent, binder, disintegrant, inclusion agent, flavoring agent, sustained-release agent, glidant, lubricant, dispersant, plasticizer, opacifier and antioxidant.

[0044] Furthermore, the dosage form of the pharmaceutical preparation is tablets, pills, capsules, powders, injections, films, lozenges, granules or oral solutions.

[0045] Furthermore, the pharmaceutical preparation further comprises one or more other drugs for preventing and / or treating myocardial injury.

[0046] Furthermore, the other drugs include coenzyme Q10, trimetazidine, coenzyme I, L-carnitine and / or creatine phosphate.

[0047] According to another aspect of the present invention, there is provided a use of the effective fraction of Herba Cynoglossi or the Herba Cynoglossi medicinal preparation in preparing a medicament for preventing and / or treating myocardial injury.

[0048] Furthermore, the effective fraction of Herba Cynoglossi or the Herba Cynoglossi medicinal preparation prevents and / or treats myocardial injury in one or more of the following ways: improving myocardial cell viability, inhibiting myocardial cell apoptosis, and promoting myocardial cell proliferation.

[0049] Furthermore, the myocardial injury is verapamil-induced myocardial injury.

[0050] Beneficial effects of the present invention:

[0051] The first effective part of the Herba Cynoglossi and / or the second effective part of the Herba Cynoglossi of the present invention can both restore human myocardial cell damage caused by verapamil, among which the first effective part of the Herba Cynoglossi has the strongest activity in promoting the overall vitality of human myocardial cells, inhibiting human myocardial cell death and promoting human myocardial cell proliferation. BRIEF DESCRIPTION OF THE DRAWINGS

[0052] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following briefly introduces the drawings required for use in the description of the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without exceeding the scope of protection required by the present invention.

[0053] Figure 1 Flow chart for the preparation of the effective parts of Herba Cynanchifoliae.

[0054] Figure 2 The following are representative images of fluorescence staining of different samples of Herba Bolognese in apoptosis assays. Green fluorescence indicates live cells stained with calcein, while red fluorescence indicates dead cells stained with PI.

[0055] Figure 3 Representative images of fluorescent staining of cells from different samples of Herba Glehniae in a cell proliferation assay. Red fluorescence indicates proliferating cells stained with Azide 555, while blue fluorescence indicates nuclei of living cells stained with Hoechst. DETAILED DESCRIPTION

[0056] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative work shall fall within the scope of protection of the present invention.

[0057] Unless otherwise indicated, all technical and scientific terms and abbreviations used herein have the meanings commonly understood by those skilled in the art in the field of the present invention or the field in which the terms are used. Although any methods, conditions, substances or materials similar or equivalent to those disclosed herein can be used in the practice of the present invention, preferred methods, conditions, substances or materials are described herein.

[0058] The present invention is intended to encompass all alternatives, modifications, and equivalents that may come within the scope of the present invention as defined by the claims. Those skilled in the art will recognize many methods and materials similar or equivalent to those described herein that could be used in the practice of the present invention. The present invention is in no way limited to the methods and materials described.

[0059] As used in the specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0060] In the present invention, the term "comprising" is synonymous with "including." As used herein, the terms "comprises," "including," "having," "containing," or any other variations thereof, are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, process, method, article, or apparatus.

[0061] As described in the background technology section, there are no reports in the prior art regarding the preparation method of the effective fraction of Herba Cynoglossi and its use in preventing and treating myocardial damage as claimed in the present invention. To address the above issues, the present invention provides a method for preparing the effective fraction of Herba Cynoglossi, comprising the following steps:

[0062] (1) extracting the herb by heating under reflux, and concentrating the extract to obtain a herb concentrate; and

[0063] (2) The Herba Glossinae concentrate is subjected to macroporous resin adsorption elution and / or polyamide resin adsorption elution to obtain the Herba Glossinae first effective fraction and / or the Herba Glossinae second effective fraction.

[0064] In a preferred embodiment, the bugloss is the dried aerial part of Anchusa italica Retz. of the Boraginaceae family.

[0065] In a preferred embodiment, the method of step (1) comprises the following steps: weighing an appropriate amount of Herba Cynanchifoliae, adding ethanol to perform a first heating reflux extraction, filtering to obtain a first filtrate and a filter residue; adding ethanol to the filter residue to perform a second heating reflux extraction, filtering to obtain a second filtrate, combining the first filtrate and the second filtrate to obtain a Herba Cynanchifoliae extract; and taking an appropriate amount of the Herba Cynanchifoliae extract, performing reduced pressure rotary evaporation, and concentrating until there is no alcohol taste, thereby obtaining the Herba Cynanchifoliae concentrate.

[0066] In a preferred embodiment, the method of step (1) does not include the step of defatting the herb.

[0067] In the present invention, when volume percent concentration, ratio, time, concentration, multiple, mass ratio, volume ratio, diameter-to-height ratio, flow rate, or other value or parameter is expressed as a range, preferred range, or a range limited by a series of upper preferred values and lower preferred values, this should be understood as specifically disclosing all ranges formed by any pairing of any range upper limit or preferred value with any range lower limit or preferred value, regardless of whether the range is disclosed separately. For example, when the range "60% to 80%" is disclosed, the described range should be interpreted as including the range "60% to 80%", "60% to 75%", "60% to 70%", "60% to 65%", "65% to 80%", "65% to 75%", "65% to 70%", "70% to 80%", "70% to 75%", "75% to 80%", etc. When a numerical range is described in this article, unless otherwise stated, the range is intended to include its end values and all integers and fractions within the range.

[0068] In a preferred embodiment, the volume percentage concentration of the ethanol is 60% to 80%, such as about 70%.

[0069] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 70%" includes ±5% of 70%, or from 66.5% to 73.5%.

[0070] In a preferred embodiment, the volume / mass (ml / g) ratio of the ethanol to the Herba Lycopodii Herba in the first heating reflux extraction is 5-15, for example, about 10.

[0071] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 10" includes ±5% of 10, or from 9.5 to 10.5.

[0072] In a preferred embodiment, the volume / mass (ml / g) ratio of the ethanol to the Herba Lycopodii Herba in the second heating reflux extraction is 4-8, for example, about 6.

[0073] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 6" includes ±5% of 6, or from 5.7 to 6.3.

[0074] In a preferred embodiment, the first heating reflux extraction time is 0.5 to 2 hours, such as about 1 hour.

[0075] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 1" includes ±5% of 1, or from 0.95 to 1.05.

[0076] In a preferred embodiment, the second heating reflux extraction time is 0.5 to 2 hours, such as about 1 hour.

[0077] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 1" includes ±5% of 1, or from 0.95 to 1.05.

[0078] In a preferred embodiment, the crude drug concentration of the Herba Cynanchifoliae concentrate is 0.1-0.5 g / ml, for example, about 0.29 g / ml.

[0079] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 0.29" includes ±5% of 0.29, or from 0.2755 to 0.3045.

[0080] In a preferred embodiment, the macroporous resin adsorption and elution method in step (2) comprises the following steps: loading the Herba Lycopersici concentrated solution onto a macroporous resin for adsorption and allowing to stand; eluting according to the following procedure: (a) water, 1 to 3 column volumes, for example, about 2 column volumes, (b) 30% to 50% ethanol, for example, about 40% ethanol, 4 to 6 column volumes, for example, about 5 column volumes, (c) 60% to 80% ethanol, for example, about 70% ethanol, 6 to 10 column volumes, for example, about 8 column volumes, and (d) 90% to 100% ethanol, for example, about 95% ethanol, 4 to 6 column volumes, for example, about 5 column volumes; collecting the elution fractions of step (b) and / or step (c), concentrating under reduced pressure, and drying to obtain the first elution fraction of Herba Lycopersici and / or the first effective fraction of Herba Lycopersici.

[0081] In the present invention, "about" refers to a value within a range of ±5% of a particular value. For example, "about 2" includes ±5% of 2, or from 1.9 to 2.1; "about 40%" includes ±5% of 40%, or from 38% to 42%; "about 5" includes ±5% of 5, or from 4.75 to 5.25; "about 70%" includes ±5% of 70%, or from 66.5% to 73.5%; "about 8" includes ±5% of 8, or from 7.6 to 8.4; "about 95%" includes ±5% of 95%, or from 90.25% to 99.75%.

[0082] In a preferred embodiment, the macroporous resin adsorption elution method further comprises a macroporous resin pretreatment step.

[0083] In a preferred embodiment, the mass ratio between the amount of the crude drug in the Herba Bovis concentrate and the amount of the macroporous resin is about 1:(1-5), for example, about 1:about 2.5.

[0084] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 1" includes ±5% of 1, or from 0.95 to 1.05; "about 2.5" includes ±5% of 2.5, or from 2.375 to 2.625.

[0085] In a preferred embodiment, the macroporous resin is a HPD 300 macroporous resin column.

[0086] In a preferred embodiment, the volume ratio of the Herba Bovis concentrate to the HPD 300 macroporous resin column is (5-9):(4-6), for example, about 7:about 5.

[0087] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 7" includes ±5% of 7, or from 6.65 to 7.35; "about 5" includes ±5% of 5, or from 4.75 to 5.25.

[0088] In a preferred embodiment, the HPD 300 macroporous resin column has a diameter-to-height ratio of about 1:about 7.

[0089] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 1" includes ±5% of 1, or from 0.95 to 1.05; "about 7" includes ±5% of 7, or from 6.65 to 7.35.

[0090] In a preferred embodiment, the flow rate of the sample loading is 0.1 to 0.5 BV / h, for example, about 0.28 BV / h.

[0091] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 0.28" includes ±5% of 0.28, or from 0.266 to 0.294.

[0092] In a preferred embodiment, the standing time is 8 to 16 hours, for example, about 12 hours.

[0093] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 12" includes ±5% of 12, or from 11.4 to 12.6.

[0094] In a preferred embodiment, the elution flow rate is 0.1 to 1 BV / h, such as about 0.7 BV / h.

[0095] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 0.7" includes ±5% of 0.7, or from 0.665 to 0.735.

[0096] In a preferred embodiment, the first effective fraction of Herba Bolognese is a fraction eluted with 60% to 80% ethanol, such as about 70% ethanol.

[0097] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 70%" includes ±5% of 70%, or from 66.5% to 73.5%.

[0098] In a preferred embodiment, the first elution portion of the Herba Bolognese is an elution portion with 30% to 50% ethanol, for example, about 40% ethanol.

[0099] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 40%" includes ±5% of 40%, or from 38% to 42%.

[0100] In a preferred embodiment, the polyamide resin adsorption elution method of step (2) comprises the following steps: dissolving the first elution fraction of Herba Cynoglossi with water, loading the fraction onto a polyamide resin for adsorption, and allowing the fraction to stand; eluting according to the following procedure: (A) water, 2 to 5 column volumes, for example, about 3.5 column volumes, (B) 50% to 70% ethanol, for example, about 60% ethanol, 5 to 10 column volumes, for example, about 7.5 column volumes, and (C) 90% to 100% ethanol, for example, about 95% ethanol, 3 to 7 column volumes, for example, about 5 column volumes; collecting the elution fraction with 50% to 70% ethanol, for example, about 60% ethanol, performing reduced pressure concentration, and drying to obtain the second effective fraction of Herba Cynoglossi.

[0101] In the present invention, "about" refers to a value within a range of ±5% of a particular value. For example, "about 3.5" includes ±5% of 3.5, or from 3.325 to 3.675; "about 60%" includes ±5% of 60%, or from 57% to 63%; "about 7.5" includes ±5% of 7.5, or from 7.125 to 7.875; "about 95%" includes ±5% of 95%, or from 90.25% to 99.75%; "about 5" includes ±5% of 5, or from 4.75 to 5.25.

[0102] In a preferred embodiment, the polyamide resin adsorption and elution method further comprises a step of pre-treating the polyamide resin.

[0103] In a preferred embodiment, the dissolution is ultrasound-assisted dissolution.

[0104] In a preferred embodiment, the volume / mass (L / g) ratio of the water to the first eluted portion of the Herba Bolognese is 0.01 to 0.1, for example, about 0.075.

[0105] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 0.075" includes ±5% of 0.075, or from 0.07125 to 0.07875.

[0106] In a preferred embodiment, the particle size of the polyamide resin is 30-60 mesh.

[0107] In a preferred embodiment, the ratio (g / L) of the mass of the first elution fraction of the Herba Bolognese to the column volume of the polyamide resin is (60-100):(1-5), for example, about 80:about 2.2.

[0108] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 80" includes ±5% of 80, or from 76 to 84; "about 2.2" includes ±5% of 2.2, or from 2.09 to 2.31.

[0109] In a preferred embodiment, the polyamide resin has an aspect ratio of about 1: about 5.5.

[0110] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 1" includes ±5% of 1, or from 0.95 to 1.05; "about 5.5" includes ±5% of 5.5, or from 5.225 to 5.775.

[0111] In a preferred embodiment, the flow rate of the sample loading is 0.1-1 BV / h, for example, about 0.65 BV / h.

[0112] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 0.65" includes ±5% of 0.65, or from 0.6175 to 0.6825.

[0113] In a preferred embodiment, the standing time is 8 to 16 hours, for example, about 12 hours.

[0114] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 12" includes ±5% of 12, or from 11.4 to 12.6.

[0115] In a preferred embodiment, the flow rate of the water elution is 0.5-1 BV / h, for example, about 0.82 BV / h.

[0116] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 0.82" includes ±5% of 0.82, or from 0.779 to 0.861.

[0117] In a preferred embodiment, the flow rate of the 50% to 70% ethanol, such as about 60% ethanol elution, is 1 to 1.5 BV / h, such as about 1.2 BV / h.

[0118] In the present invention, "about" refers to a value within the range of ±5% of a specific value. For example, "about 60%" includes ±5% of 60%, or from 57% to 63%; "about 1.2" includes ±5% of 1.2, or from 1.14 to 1.26.

[0119] In a preferred embodiment, the flow rate of the 90% to 100% ethanol, such as about 95% ethanol elution, is 1 to 3 BV / h, such as about 1.9 BV / h.

[0120] In the present invention, "about" refers to a value within the range of ±5% of a specific value. For example, "about 95%" includes ±5% of 95%, or from 90.25% to 99.75%; "about 1.9" includes ±5% of 1.9, or from 1.805 to 1.995.

[0121] According to another aspect of the present invention, an effective part of Herba Cynoglossi prepared by the above-mentioned preparation method is provided.

[0122] The effective parts of the Herba Lycopersicum var. ameloides include the first effective part of the Herba Lycopersicum var. ameloides and / or the second effective part of the Herba Lycopersicum var. ameloides.

[0123] According to another aspect of the present invention, a Herba Cynoglossi pharmaceutical preparation is provided, which comprises the effective part of the Herba Cynoglossi and one or more pharmaceutically acceptable excipients.

[0124] In a preferred embodiment, the excipient is selected from one or more of the following: diluents, wetting agents, binders, disintegrants, inclusion agents, flavoring agents, sustained-release agents, glidants, lubricants, dispersants, plasticizers, opacifiers and antioxidants.

[0125] In a preferred embodiment, the pharmaceutical preparation is in the form of tablets, pills, capsules, powders, injections, films, lozenges, granules or oral solutions.

[0126] In a preferred embodiment, the pharmaceutical preparation further comprises one or more other drugs for preventing and / or treating myocardial injury.

[0127] In a preferred embodiment, the other drugs include coenzyme Q10, trimetazidine, coenzyme I, L-carnitine and / or creatine phosphate.

[0128] According to another aspect of the present invention, there is provided a use of the effective fraction of Herba Cynoglossi or the Herba Cynoglossi medicinal preparation in preparing a medicament for preventing and / or treating myocardial injury.

[0129] In a preferred embodiment, the effective fraction of Herba Cynoglossi or the Herba Cynoglossi pharmaceutical preparation prevents and / or treats myocardial injury in one or more of the following ways: improving myocardial cell viability, inhibiting myocardial cell apoptosis, and promoting myocardial cell proliferation.

[0130] In a preferred embodiment, the myocardial injury is verapamil-induced myocardial injury.

[0131] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the invention. The experimental methods in the following examples, for which specific conditions are not specified, are generally based on conventional conditions or conditions recommended by the manufacturer.

[0132] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those familiar to those skilled in the art. Furthermore, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention. The preferred embodiments and materials described herein are for illustrative purposes only.

[0133] The above-mentioned features of the present invention or the features described in the embodiments may be combined in any combination. All features disclosed in this patent specification may be used in any combination, and each feature disclosed in the specification may be replaced by any alternative feature that can provide the same, equal, or similar purpose. Therefore, unless otherwise specified, the features disclosed are only general examples of equal or similar features.

[0134] Example

[0135] Preparation Example

[0136] 1. Instruments and Materials

[0137] 1.1 Instruments

[0138] ZNHW-10L intelligent digital display electric heating mantle (Tianjin Yuhua Instrument Technology Co., Ltd.); KQ-250DB CNC ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.); TD20001A electronic balance (Tianjin Tianma Hengji Instrument Co., Ltd.); EYELA rotary evaporator (model: SB-1100) purchased from Shanghai Ailang Instrument Co., Ltd.; CT11-HH-6 constant temperature water bath (Beijing Haifuda Technology Co., Ltd.); GZX-9070MB electric blast drying oven (Shanghai Boxun Industrial Co., Ltd. Medical Equipment Factory).

[0139] 1.2 Experimental Materials

[0140] Ox tongue herb (batch number: C202312002) was purchased from Hebei Jiaheng Lengbei Pharmaceutical Co., Ltd.; HPD300 macroporous adsorption resin was purchased from Solebau Technology Co., Ltd.; 30-60 mesh polyamide resin was purchased from Yuanbaoshan Chromatography Technology Co., Ltd.; 95% ethanol was purchased from Meihekou Fukang Alcohol Co., Ltd.; water was distilled water.

[0141] 1.3 Preparation method of total extract of Herba Lycopodii

[0142] Take about 2.5 kg of Herba Cynoglossi, add 10 times the amount of 70% ethanol, heat and reflux to extract for 1 hour, filter, add 6 times the amount of 70% ethanol to the residue, heat and reflux to extract for 1 hour, filter, and combine the two filtrates to obtain Herba Cynoglossi extract. Among them, take 1 / 5 of the extract, concentrate under reduced pressure, and dry to obtain Herba Cynoglossi total extract (ZTW, about 93.7 g); take another 4 / 5 of the extract, vacuum rotary evaporation, and concentrate until there is no alcohol taste, to obtain Herba Cynoglossi top concentrate (about 7.0 L, equivalent to 0.29 g crude drug / mL).

[0143] 1.4. Preparation method of each elution part of macroporous resin

[0144] The concentrated aqueous solution of Herba Bovis was loaded onto a pretreated HPD 300 macroporous resin column (crude drug amount: resin amount = 1:2.5), with a column volume of 5 L and a diameter-to-height ratio of 1:7. The loading flow rate was approximately 0.28 BV / h, and the fraction during the loading process was the unadsorbed fraction. After the loading was completed, the column was allowed to stand for 12 h, and 2 BV of water, 5 BV of 40% ethanol, 8 BV of 70% ethanol, and 5 BV of ethanol were used in sequence. The product was eluted with 95% ethanol in sequence at a rate of about 0.7 BV / h. The water elution fraction, 40% ethanol elution fraction, 70% ethanol elution fraction and 95% ethanol elution fraction were collected respectively. The collected fractions were concentrated under reduced pressure and dried to obtain the non-adsorbed portion (D-1, 141.70 g) of HPD300, the water-washed portion (D-2, 93.67 g), the 40% ethanol elution portion (D-3, 107.1 g), the 70% ethanol elution portion (D-4, 30.06 g) and the 95% ethanol elution portion (D-5, 4.12 g).

[0145] 1.5. Preparation methods of various elution sites of polyamide resin

[0146] Take 80 g of powder from the 40% ethanol elution portion of the above-mentioned macroporous resin, add water and ultrasonically dissolve it (about 6.0 L), and load it onto the pretreated 30-60 mesh polyamide resin. The column volume is 2.2 L, the diameter-to-height ratio is 1:5.5, the loading flow rate is about 0.65 BV / h, and the first 0.5 BV of effluent is discarded. The remaining fractions during the loading process are unadsorbed fractions. After the loading is completed, let it stand for 12 hours, and use 3.5 BV of water, 7.5 BV of 60% ethanol, and 5.0 BV of 95% ethanol to elute in sequence. The elution rates are 0.82 BV / h, 1.20 BV / h, and 1.90 BV / h, respectively, to obtain water elution fractions, 60% ethanol elution fractions, and 95% ethanol elution fractions, respectively. The collected fractions were concentrated under reduced pressure and dried to obtain the non-adsorbed portion of the polyamide resin (J-1, 26.11 g), the water-washed portion (J-2, 20.17 g), the 60% ethanol elution portion (J-3, 23.29 g) and the 95% ethanol elution portion (J-4, 1.20 g), respectively.

[0147] The flow chart for preparing the effective parts of Herba Cynanchifoliae is as follows Figure 1 shown.

[0148] Efficacy Examples

[0149] 1. Instruments and Materials

[0150] 1.1 Cell lines

[0151] AC16 human cardiomyocytes (CL-0790) were purchased from Wuhan Punosai Life Science Co., Ltd. Cell culture conditions: gas phase: air, 95%; CO2, 5%; temperature: 37°C.

[0152] 1.2 Experimental Instruments

[0153] The CO2 incubator (model: SERIES 8000WJ) was purchased from Thermo Fisher Scientific Inc.; the cell counter (model: AXC1100509) was purchased from Olympus, Japan; the microplate reader (model: SPECTROstar Nano) was purchased from BMG LABTECH; the high-speed centrifuge (model: ST16) was purchased from Thermo Fisher Scientific Inc.; the EYELA rotary evaporator (model: SB-1100) was purchased from Shanghai Ailan Instrument Co., Ltd.; and the New Century UV-visible spectrophotometer (model: T6) was purchased from Shanghai Yuanxi Instrument Co., Ltd.

[0154] 1.3 Experimental Materials

[0155] Verapamil hydrochloride (Ver, batch number: D23231227) was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.; DEME / F12 (batch number: 6124490) was purchased from Thermo Fisher Scientific; fetal bovine serum (batch number: FBS1612-109) was purchased from BIOEXPLORER; penicillin-streptomycin solution (double antibody) (batch number: PB180120) was purchased from Wuhan Punosai Life Science Technology Co., Ltd.; trypsin solution (batch number: PB180225) and Cell Counting Kit-8 (CCK-8) reagent (batch number: SK2060) were purchased from Beijing Coolbo Technology Co., Ltd.; Calcein / PI cell viability and cytotoxicity detection kit (batch number: C2015S) and BeyoClick TM EdU-555 cell proliferation detection kit (lot number: C0075S), immunofluorescence fixative (lot number: P0098), immunofluorescence washing solution (lot number: P0106), immunofluorescence permeabilization solution (lot number: P0095), and Hoechst 33342 live cell staining solution (lot number: C1027) were all purchased from Beyotime Biotechnology Co., Ltd.

[0156] 2. Methods and Results

[0157] 2.1. Preparation of drug solution

[0158] Take appropriate amounts of the total extract of Herba Cynanchifoliae (ZTW) prepared according to the method of "1.3", the macroporous resin unadsorbed fraction (D-1), the macroporous resin water-washed fraction (D-2), the macroporous resin 40% ethanol eluted fraction (D-3), the macroporous resin 70% ethanol eluted fraction (D-4), the macroporous resin 95% ethanol eluted fraction (D-5) under "1.4", and the polyamide resin unadsorbed fraction (J-1), the polyamide resin water-washed fraction (J-2), the polyamide resin 60% ethanol eluted fraction (J-3), and the polyamide resin 95% ethanol eluted fraction (J-4) under "1.5", accurately weigh them, dissolve them in DMSO and prepare 0.25 mg mL -1 The solution was diluted to 62.5 μg·mL with basal culture medium. -1 The molecular formula of verapamil hydrochloride is C 27 H 38 N2O4, with a relative molecular mass of 491.06, accurately weigh 0.0200g of Ver, dissolve it in 509μL of basal culture medium, and shake to mix to obtain a Ver mother solution with a concentration of 80mM. Pipette a certain volume of the mother solution into a centrifuge tube and dilute it with basal culture medium to the corresponding dosing concentration.

[0159] 2.2 Cell culture and passaging

[0160] 2.2.1 Cell recovery

[0161] Before cell recovery, preheat the water bath to 37°C and add the prepared complete medium (15% FBS + 1% double-antibody + 84% DMEM / F12) to a 15mL centrifuge tube in advance. Remove the AC16 cells to be recovered from the -80°C freezer and quickly thaw them in a 37°C water bath. After thawing, quickly transfer the cells from the cryopreserved tube to a centrifuge tube that has been previously infused with complete medium in a clean bench. Centrifuge at 800rpm for 5 minutes. After centrifugation, remove the supernatant, add complete medium to the centrifuge tube, pipette evenly, and transfer the cell suspension to a cell culture flask. Place the cell culture flask in a 37°C, 5% CO2 cell culture incubator.

[0162] 2.2.2 Cell Passaging

[0163] Observe the cells under a microscope. When the cell density reaches 80-90%, remove the culture medium from the culture flask, add 1 mL of PBS to wash twice, and add 1 mL of trypsin solution. When the cell morphology is observed to become round under an optical microscope, add complete culture medium to the culture flask to terminate digestion and pipette until the cells are completely suspended in the culture medium. Transfer the cell suspension to a 15 mL centrifuge tube and centrifuge at 800 rpm for 5 minutes. After centrifugation, remove the supernatant, add complete culture medium to the centrifuge tube, pipette evenly, and transfer the cell suspension to a cell culture flask. Place the cell culture flask in a 37°C, 5% CO2 cell culture incubator.

[0164] 2.2.3 Cell Cryopreservation

[0165] Prepare a cell suspension of adherent cells by referring to the method in “2.2.2”, add freezing solution (55% DMEM / F12 + 40% FBS + 5% DMSO) into the centrifuge tube, pipette evenly, transfer the cell suspension into a cryovial, place the cryovial in a freezing box, and quickly transfer to -80°C.

[0166] 2.3 Effects on cell viability

[0167] AC16 cells in the logarithmic growth phase were seeded in 96-well plates at a cell density of 7.5 × 10 4 The experiment was conducted using a blank group, a Ver group, and Ver + Herba Lycopersici total extract or different elution fraction groups. After adding the test drug to each well, the plate was incubated in a 37°C, 5% CO2 cell culture incubator for 4.5 hours. The supernatant was discarded after the incubation. A certain concentration of Ver was then added to the Ver + Herba Lycopersici total extract or different elution fraction groups. The plate was incubated in a 37°C, 5% CO2 cell culture incubator for 30 minutes. After the administration, the supernatant was discarded, and the cells were washed twice with PBS. 100 μL of DMEM / F12 basal medium containing 10% CCK-8 solution was added to the wells and incubated in an incubator for 2 hours. After the incubation, the OD value at 450 nm was measured using a multifunctional microplate reader, and the cell survival rate of the corresponding group was calculated.

[0168] As shown in Table 1, compared with the blank group, the cell viability of the Ver group was significantly decreased (P<0.0001). Compared with the Ver group, the concentration of the sample group was 62.5 μg·mL -1Under the conditions of 62.5 μg·mL, the cell viability of the D-3 sample group increased significantly (P<0.05), and the cell viability of the D-4 sample group and the J-3 sample group increased more significantly (P<0.0001). There were no significant differences in the other sample groups. In addition, compared with the J-3 sample group, the cell viability of the D-4 sample group increased more significantly (P<0.001). The above results show that compared with the total extract of Herba Botrytis cava and other elution fractions, the J-3 sample group and the D-4 sample group had a higher cell viability than the Herba Botrytis cava total extract and other elution fractions at a concentration of 62.5 μg·mL -1 When the drug was added to the sample, the overall viability of cardiomyocytes was significantly promoted, and the activity of the D-4 sample group was the strongest. Compared with the Ver group, the D-1 sample group, D-2 sample group, D-5 sample group, J-2 sample group, and J-4 sample group even showed a trend of further decline in cardiomyocyte viability.

[0169] Table 1 Effects of Herba Lycopodii Total Extract and Different Elution Fractions on Cell Viability ( n=4)

[0170] Group concentration Cell viability (%) Blank group - 100±1.24 Ver Group 200 μM 81.9±2.20#### Ver+ZTW <![CDATA[62.5μg·mL -1 ]]> 82.4±1.98 Ver+D-1 <![CDATA[62.5μg·mL -1 ]]> 80.1±3.08 Ver+D-2 <![CDATA[62.5μg·mL -1 ]]> 81.2±2.79 Ver+D-3 <![CDATA[62.5μg·mL -1 ]]> 85.9±1.75* Ver+D-4 <![CDATA[62.5μg·mL -1 ]]> 95.1±1.54**** Ver+D-5 <![CDATA[62.5μg·mL -1 ]]> 78.7±2.43 Ver+J-1 <![CDATA[62.5μg·mL -1 ]]> 82.0±3.20 Ver+J-2 <![CDATA[62.5μg·mL -1 ]]> 80.9±1.75 Ver+J-3 <![CDATA[62.5μg·mL -1 ]]> 90.2±1.32**** Ver+J-4 <![CDATA[62.5μg·mL -1 ]]> 79.7±2.04

[0171] Note: # indicates the comparison between Ver group and blank group, * indicates the comparison between different sample groups and Ver group, **** / #### P<0.0001, *P<0.05.

[0172] 2.4 Effects on human cardiomyocyte apoptosis

[0173] The experiment included blank, Ver, Ver+J-3, and Ver+D-4 groups. After adding the corresponding concentrations of the test drug to the wells, the plates were incubated in a 37°C, 5% CO2 cell culture incubator for 4.5 hours. The supernatant was discarded after the incubation period. Ver was then added to the Ver+J-3 and Ver+D-4 groups at a specific concentration. The plates were incubated in a 37°C, 5% CO2 cell culture incubator for 30 minutes. After the administration period, the supernatant was discarded and the plates were washed twice with PBS. A specific volume of calcein / propidium iodide working solution was prepared, 100 μL of which was added to each well, and the cells were incubated at 37°C in the dark for 30 minutes. After the incubation period, images were taken using a high-content imaging system, and Image J software was used to count the number of dead cells and total cells, and calculate cell mortality.

[0174] As shown in Table 2 and Figure 2 As shown in the results, compared with the blank group, the cell death rate in the Ver group was significantly increased (P<0.0001). Compared with the Ver group, the cell death rates in the J-3 sample group and the D-4 sample group were significantly reduced (P<0.001, P<0.0001). In addition, compared with the J-3 sample group, the cell death rate in the D-4 sample group was significantly reduced (P<0.01). The above results show that the J-3 sample group and the D-4 sample group have a significant effect on the cell death rate at a concentration of 62.5 μg·mL-1 When the cells were treated with D-4, the myocardial cell death rate was significantly reduced, and the activity of the D-4 sample group was stronger.

[0175] Table 2 Effects of different samples of Herba Bolognese on cell apoptosis ( n=4)

[0176] Group concentration Cell death rate (%) Blank control group - 5.74±1.70 Ver Group 200 μM <![CDATA[18.4±4.47 #### ]]> Ver+J-3 group <![CDATA[62.5μg·mL -1 ]]> <![CDATA[8.48±2.38 *** ]]> Ver+D-4 group <![CDATA[62.5μg·mL -1 ]]> <![CDATA[3.30±0.467 **** ]]>

[0177] # indicates the comparison between the model group and the blank group, * indicates the comparison between different sample groups and the model group, **** / #### P < 0.0001, *** P<0.001.

[0178] 2.5 Effects on Human Cardiomyocyte Proliferation

[0179] The experiment consisted of blank, Ver, Ver+J-3, and Ver+D-4 groups. After adding the test drug at the appropriate concentration to the wells, the plates were incubated in a 37°C, 5% CO2 cell culture incubator for 4.5 hours. The supernatant was discarded after the incubation period. Ver was then added to the Ver+J-3 and Ver+D-4 groups at the appropriate concentrations. The plates were incubated in a 37°C, 5% CO2 cell culture incubator for 30 minutes. After the administration period, the supernatant was discarded and the cells were washed twice with PBS. A volume of 5-ethynyl-2'-deoxyuridine (EdU) working solution (1×) was prepared and heated to 37°C. 100 μL of EdU working solution was added to each well and incubated at 37°C for an additional 2 hours. After the incubation period, the supernatant was discarded, and 100 μL of fixative solution was added to each well. The cells were fixed at room temperature for 15 minutes. The fixative solution was discarded, and the cells were washed three times with 100 μL of wash solution per well for 3-5 minutes each. Discard the wash solution, add 100 μL of permeabilization solution to each well, incubate at room temperature for 10-15 minutes, discard the permeabilization solution, and wash the cells one to two times with 100 μL of wash solution per well for 3-5 minutes each. Prepare a volume of Click reaction solution and add 100 μL of Click reaction solution to each well. Incubate at room temperature in the dark for 30 minutes, then discard the solution and wash three times with wash solution for 3-5 minutes each. Discard the wash solution and add 100 μL of Hoechst live cell stain (1×) to each well. Incubate at room temperature in the dark for 10 minutes, then discard the Hoechst live cell stain and wash three times with 100 μL of wash solution per well for 3-5 minutes each. Images were taken using a high-content imaging system. Image J software was used to count the number of proliferating cells and total cells and calculate the cell proliferation rate.

[0180] As shown in Table 3 and Figure 3As shown in the figure, compared with the blank group, the cell proliferation rate of the Ver group was significantly reduced (P<0.001). Compared with the Ver group, the cell proliferation rates of the J-3 sample group and the D-4 sample group were significantly increased (P<0.05). The above results show that the J-3 sample group and the D-4 sample group have a significant effect on the cell proliferation rate of the Ver group at a concentration of 62.5μg·mL -1 The proliferation rate of cardiomyocytes can be significantly increased.

[0181] Table 3 Effects of different samples of Herba Bolognese on cell proliferation ( n=4)

[0182]

[0183] # indicates the comparison between the model group and the blank group, * indicates the comparison between different sample groups and the model group, ### P<0.001, * P<0.05.

[0184] 3. Conclusion

[0185] Compared with the total extract of Ampelopsis ophiopogonis and other elution fractions, the polyamide resin 60% ethanol elution fraction (J-3) and the macroporous resin 70% ethanol elution fraction (D-4) can significantly promote the overall cell viability of human cardiomyocytes, while inhibiting the apoptosis of human cardiomyocytes and promoting the cell proliferation of human cardiomyocytes, indicating that J-3 and D-4 have good effects on restoring human cardiomyocyte damage caused by verapamil.

[0186] The embodiments of the present invention are described in detail above. Specific examples are used herein to illustrate the principles and implementation methods of the present invention. The description of the above embodiments is only intended to help understand the method of the present invention and its core concept. At the same time, changes or modifications made by those skilled in the art based on the concept of the present invention, the specific implementation methods of the present invention, and the scope of application are all within the scope of protection of the present invention. In summary, the contents of this specification should not be understood as limiting the present invention.

Claims

1. A method for preparing the effective part of Herba Cynanchifoliae, characterized in that: The preparation method comprises the following steps: (1) extracting the herb by heating under reflux, and concentrating the extract to obtain a herb concentrate; and (2) The Herba Glossinae concentrate is subjected to macroporous resin adsorption elution and / or polyamide resin adsorption elution to obtain the Herba Glossinae first effective fraction and / or the Herba Glossinae second effective fraction.

2. The preparation method according to claim 1, characterized in that The Anchusa italica Retz is the dried aerial part of the Boraginaceae family Anchusa italica Retz.

3. The preparation method according to claim 1, characterized in that The method of step (1) comprises the following steps: weighing an appropriate amount of Herba Cynanchii, adding ethanol to perform a first heating reflux extraction, filtering to obtain a first filtrate and a filter residue; adding ethanol to the filter residue to perform a second heating reflux extraction, filtering to obtain a second filtrate, combining the first filtrate and the second filtrate to obtain a Herba Cynanchii extract; and taking an appropriate amount of the Herba Cynanchii extract, performing reduced pressure rotary evaporation, and concentrating until there is no alcohol taste, to obtain the Herba Cynanchii concentrate; Preferably, the method of step (1) does not include the step of defatting the herb. Preferably, the volume percentage concentration of the ethanol is 60% to 80%, for example, about 70%; Preferably, the volume / mass (ml / g) ratio of the ethanol to the Herba Lycopodii in the first heating reflux extraction is 5 to 15, for example, about 10; Preferably, the volume / mass (ml / g) ratio of the ethanol to the Herba Lycopodii Herba in the second heating reflux extraction is 4 to 8, for example, about 6; Preferably, the first heating reflux extraction time is 0.5 to 2 hours, for example, about 1 hour; Preferably, the second heating reflux extraction time is 0.5 to 2 hours, for example, about 1 hour; Preferably, the crude drug concentration of the Herba Lycopodii Herba Concentrate is 0.1-0.5 g / ml, for example, about 0.29 g / ml.

4. The preparation method according to claim 1, characterized in that The macroporous resin adsorption and elution method of step (2) comprises the following steps: loading the Herba Cynoglossi concentrated solution onto a macroporous resin for adsorption and allowing to stand; eluting according to the following procedure: (a) water, 1 to 3 column volumes, for example, about 2 column volumes, (b) 30% to 50% ethanol, for example, about 40% ethanol, 4 to 6 column volumes, for example, about 5 column volumes, (c) 60% to 80% ethanol, for example, about 70% ethanol, 6 to 10 column volumes, for example, about 8 column volumes, and (d) 90% to 100% ethanol, for example, about 95% ethanol, 4 to 6 column volumes, for example, about 5 column volumes; collecting the elution fractions of step (b) and / or step (c), concentrating under reduced pressure, and drying to obtain the Herba Cynoglossi first elution fraction and / or the Herba Cynoglossi first effective fraction; Preferably, the macroporous resin adsorption and elution method further comprises the step of pre-treating the macroporous resin; Preferably, the mass ratio between the amount of the crude drug in the Herba Bovis concentrate and the amount of the macroporous resin is about 1:(1-5), for example, about 1:about 2.5; Preferably, the macroporous resin is a HPD 300 macroporous resin column; Preferably, the volume ratio between the Herba Bovis concentrate and the HPD 300 macroporous resin column is (5-9):(4-6), for example, about 7:about 5; Preferably, the diameter-to-height ratio of the HPD 300 macroporous resin column is about 1: about 7; Preferably, the flow rate of the sample loading is 0.1 to 0.5 BV / h, for example, about 0.28 BV / h; Preferably, the standing time is 8 to 16 hours, for example, about 12 hours; Preferably, the elution flow rate is 0.1 to 1 BV / h, for example, about 0.7 BV / h; More preferably, the first effective part of the Herba Bolognese is a part eluted with 60% to 80% ethanol, such as about 70% ethanol; More preferably, the first elution portion of the Herba Bolognese is an elution portion with 30% to 50% ethanol, such as about 40% ethanol.

5. The preparation method according to claim 1, characterized in that The polyamide resin adsorption and elution method of step (2) comprises the following steps: dissolving the first elution fraction of the Herba Bolognese with water, applying the sample to the polyamide resin for adsorption, and allowing to stand; eluting according to the following procedure: (A) water, 2 to 5 column volumes, for example, about 3.5 column volumes, (B) 50% to 70% ethanol, for example, about 60% ethanol, 5 to 10 column volumes, for example, about 7.5 column volumes, and (C) 90% to 100% ethanol, for example, about 95% ethanol, 3 to 7 column volumes, for example, about 5 column volumes; collecting the elution fraction with 50% to 70% ethanol, for example, about 60% ethanol, performing reduced pressure concentration, and drying to obtain the second effective fraction of the Herba Bolognese; Preferably, the polyamide resin adsorption and elution method further comprises the step of pre-treating the polyamide resin; Preferably, the dissolution is ultrasound-assisted dissolution; Preferably, when water is added for dissolution, the volume / mass (L / g) ratio of the water to the first eluted portion of the Herba Glechomae is 0.01 to 0.1, for example, about 0.075; Preferably, the particle size of the polyamide resin is 30 to 60 mesh; Preferably, the ratio (g / L) of the mass of the first elution portion of the Herba Bolognese to the column volume of the polyamide resin is (60-100):(1-5), for example, about 80:about 2.2; Preferably, the polyamide resin has a diameter-to-height ratio of about 1: about 5.5; More preferably, the flow rate of the sample loading is 0.1 to 1 BV / h, for example, about 0.65 BV / h; More preferably, the standing time is 8 to 16 hours, for example about 12 hours; More preferably, the flow rate of the water elution is 0.5 to 1 BV / h, for example, about 0.82 BV / h; More preferably, the flow rate of the 50% to 70% ethanol, for example, about 60% ethanol elution, is 1 to 1.5 BV / h, for example, about 1.2 BV / h; More preferably, the flow rate of the 90% to 100% ethanol, such as about 95% ethanol elution, is 1 to 3 BV / h, such as about 1.9 BV / h.

6. An effective part of Herba Cynanchifoliae prepared by the method according to any one of claims 1 to 5; in, The effective parts of Herba Cynoglossi include the first effective part of Herba Cynoglossi and / or the second effective part of Herba Cynoglossi.

7. A medicinal preparation of Herba Bolognese, characterized in that: The pharmaceutical preparation comprises the effective part of Herba Bolognese as claimed in claim 6, and one or more pharmaceutically acceptable excipients; Preferably, the excipient is selected from one or more of the following: diluent, wetting agent, binder, disintegrant, inclusion agent, flavoring agent, sustained-release agent, glidant, lubricant, dispersant, plasticizer, opacifier and antioxidant; More preferably, the dosage form of the pharmaceutical preparation is tablet, pill, capsule, powder, injection, film, lozenge, granule or oral solution.

8. The pharmaceutical preparation according to claim 7, characterized in that The pharmaceutical preparation further comprises one or more other drugs for preventing and / or treating myocardial damage; Preferably, the other drugs include coenzyme Q10, trimetazidine, coenzyme I, L-carnitine and / or creatine phosphate.

9. Use of the effective fraction of Herba Bolognese according to claim 6 or the Herba Bolognese pharmaceutical preparation according to claim 7 or 8 in the preparation of a medicament for preventing and / or treating myocardial injury.

10. The use according to claim 9, characterized in that The effective part of the Herba Bolognese or the Herba Bolognese medicinal preparation prevents and / or treats myocardial injury in one or more of the following ways: improving myocardial cell viability, inhibiting myocardial cell apoptosis and promoting myocardial cell proliferation; Preferably, the myocardial injury is verapamil-induced myocardial injury.

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