CN116889589A

CN116889589A - Chinese herbal compound preparation for treating sepsis ARDS

Info

Publication number: CN116889589A
Application number: CN202310954840.0A
Authority: CN
Inventors: 方邦江; 周爽; 彭伟; 张文; 吴霖光缙; 徐湘茹; 郑月娟; 屠亦文; 王蓓
Original assignee: Longhua Hospital Affiliated to Shanghai University of TCM
Current assignee: Longhua Hospital Affiliated to Shanghai University of TCM
Priority date: 2023-08-01
Filing date: 2023-08-01
Publication date: 2023-10-17

Abstract

The invention discloses a traditional Chinese medicine compound preparation for treating sepsis ARDS, belonging to the field of traditional Chinese medicine formulas. The traditional Chinese medicine compound preparation comprises the following components in parts by weight: 50-60 parts of ginseng root, 40-45 parts of rheum officinale stem, 30-35 parts of sargentgloryvine stem, 30-35 parts of dandelion, 50-55 parts of aconite and 6-10 parts of leech. Clinical experiments prove that the prepared traditional Chinese medicine compound preparation contains various active ingredients, can regulate and control TWIK2-NLRP3 signal paths, can obviously improve related symptoms of sepsis ARDS, can obviously reduce the death rate of patients, can effectively delay the progress of illness, and can greatly reduce the invasive mechanical ventilation proportion of the patients. The invention provides a feasible new scheme for treating sepsis ARDS diseases, and has practical clinical significance and popularization value.

Description

Chinese herbal compound preparation for treating sepsis ARDS

Technical Field

The invention relates to the field of traditional Chinese medicine formulas, in particular to a traditional Chinese medicine compound preparation for treating sepsis ARDS.

Background

Clinical acute respiratory distress syndrome (Acute respiratory distress syndrome, ARDS) is one of the most serious complications of sepsis, and is also an independent risk factor for increased mortality, hospitalization time and prolonged mechanical ventilation time in sepsis patients, which has been a difficult point of clinical sepsis treatment. For ARDS patients, support therapy is often clinically used, including mechanical ventilation, prevention of stress ulcers and venous thromboembolism, nutritional support and treatment of potential injury, whereas mechanical ventilation is the only effective measure of sepsis with ARDS, although to some extent improving the prognosis of the patient, the incidence and mortality of sepsis with ARDS is still high, presenting a great challenge to clinical treatment and also a heavy burden to society and families.

Although the concept of sepsis is not seen in classical collection of traditional Chinese medicine, modern doctors can classify the disease into diseases such as typhoid fever, exogenous febrile disease, warm toxin, yellow, invagination and the like due to symptoms such as fever, syncope and unsmooth viscera qi. Clinical observation by modern doctors shows that the syndrome elements of sepsis combined with ARDS mainly comprise heat toxin, phlegm, blood stasis and deficiency, and the onset of the sepsis is easy to be deficiency and excess, and then the sepsis is often manifested as deficiency-excess mixed syndrome which is caused by deficiency of healthy qi and deficiency of pathogenic qi.

Therefore, for the clinical refractory disease, active intervention of traditional Chinese medicine is needed to develop effective medicine and method research for treating sepsis complicated with ARDS.

Disclosure of Invention

The invention aims to provide a traditional Chinese medicine compound preparation for treating sepsis ARDS, which solves the problems in the prior art, and the traditional Chinese medicine compound preparation (Shenhuang particles) prepared by the invention can regulate TWIK2-NLRP3 signal paths, can obviously improve related symptoms of sepsis ARDS, reduce the death rate of patients, can also effectively delay the progress of illness, greatly reduce the invasive mechanical ventilation proportion of the patients, provides a practical new scheme for treating sepsis ARDS diseases, and has practical clinical significance and popularization value.

In order to achieve the above object, the present invention provides the following solutions:

the invention provides a traditional Chinese medicine compound preparation for treating sepsis ARDS, which comprises the following components in parts by weight:

50-60 parts of ginseng root, 40-45 parts of rheum officinale stem, 30-35 parts of sargentgloryvine stem, 30-35 parts of dandelion, 50-55 parts of aconite and 6-10 parts of leech.

Further, the paint comprises the following components in parts by weight:

50 parts of ginseng root, 40 parts of rheum officinale stem, 30 parts of sargentgloryvine stem, 30 parts of dandelion, 50 parts of aconite and 6 parts of leech.

The invention also provides a preparation method of the traditional Chinese medicine compound preparation, which comprises the following steps:

weighing the medicinal materials according to the parts by weight, crushing and mixing uniformly, carrying out ethanol heating reflux extraction to obtain an extracting solution, removing ethanol in the extracting solution, concentrating and drying to obtain the traditional Chinese medicine compound preparation.

Further, the ethanol is an ethanol solution with the concentration of 85vol%, and the volume ratio of the medicinal material to the ethanol is 1: (8-10); the conditions of the ethanol heating reflux extraction are as follows: heating at 80deg.C, and reflux extracting for 2-3 times.

The invention also provides application of the traditional Chinese medicine compound preparation in preparing medicines for treating sepsis ARDS.

Furthermore, the Chinese herbal compound preparation can improve symptoms of patients with sepsis ARDS, reduce mortality and delay disease progression by regulating TWIK2-NLRP3 signal paths, and has the effect of treating sepsis ARDS.

Further, the medicament also comprises pharmaceutically acceptable auxiliary materials.

Further, the pharmaceutically acceptable auxiliary materials comprise any one or more of a filler, a disintegrating agent, an adhesive, a lubricant and a flavoring agent.

The invention discloses the following technical effects:

the invention discloses a self-simulating Chinese herbal compound ginseng-astragalus granule by utilizing the concept of 'cut-off reversion', which consists of ginseng, rhubarb, sargentgloryvine stem, dandelion, aconite and leech. In the formula, ginseng is used for supplementing primordial qi as a monarch, aconite is used for strengthening body resistance and restoring yang for rescuing collapse, and Huang Tongfu is used for clearing lung and directly relieving heat potential, and sargentgloryvine stem and dandelion are used for clearing heat and detoxicating, and leech is used for promoting blood circulation and removing blood stasis and resolving dampness. The whole formula has the effects of restoring yang, rescuing from collapse, removing blood stasis, resolving dampness, clearing heat and detoxicating.

Clinical experiments prove that the prepared ginseng-astragalus particles contain various active ingredients, can regulate and control TWIK2-NLRP3 signal paths, can obviously improve related symptoms of sepsis ARDS, can obviously reduce the death rate of patients, can effectively delay the progress of illness, and can greatly reduce the invasive mechanical ventilation proportion of the patients. The invention provides a feasible new scheme for treating sepsis ARDS diseases, and has practical clinical significance and popularization value.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.

FIG. 1 is a ginseng yellow particle-based peak ion flow diagram; a: a positive ion mode; b: a negative ion mode;

FIG. 2 is a molecular docking model; a: ginsenoside Re; b: arctiin; c: benzoyl aconitine; d: benzoylmesaconine; e: benzoyl mesaconitine; f: aconitine; g: chicoric acid; h: phenylalanine; i: new chlorogenic acid; j: chlorogenic acid;

FIG. 3 is a model establishment of LPS-induced sepsis ARDS mice; a: normal mice; b: sepsis ARDS mice;

FIG. 4 is a graph of the improvement of lung tissue injury in sepsis ARDS mice with ginseng-yellow particles; a: lung tissue injury scoring results; b: lung wet/dry weight ratio results; c: paO2/FiO2 ratio results;

FIG. 5 is a clinical efficacy of Shenhuang granule in improving sepsis ARDS; a: baseline and post-treatment severity; b: mortality at points;

FIG. 6 is a clinical efficacy of Shenhuang granule in improving sepsis ARDS; a: rate of disease progression; b: invasive mechanical ventilation ratios are used.

Detailed Description

Various exemplary embodiments of the invention will now be described in detail, which should not be considered as limiting the invention, but rather as more detailed descriptions of certain aspects, features and embodiments of the invention.

It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In addition, for numerical ranges in this disclosure, it is understood that each intermediate value between the upper and lower limits of the ranges is also specifically disclosed. Every smaller range between any stated value or stated range, and any other stated value or intermediate value within the stated range, is also encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.

It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the invention described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from consideration of the specification of the present invention. The specification and examples of the present invention are exemplary only.

As used herein, the terms "comprising," "including," "having," "containing," and the like are intended to be inclusive and mean an inclusion, but not limited to.

Example 1 Chinese herbal Compound preparation for treating sepsis ARDS

1 Experimental materials

The ginseng root, the rheum officinale stem, the sargentgloryvine stem, the dandelion whole plant, the aconite root and the leech whole plant are purchased from a traditional Chinese medicine pharmacy of an affiliated Longhua hospital at Shanghai traditional Chinese medicine university.

2 pharmaceutical composition of Chinese medicinal compound preparation (SHENHUANG granule)

The composition comprises the following components:

50g of ginseng root, 40g of rheum officinale stem, 30g of sargentgloryvine stem, 30g of dandelion whole plant, 50g of aconite root and 6g of leech whole plant.

3 preparation process

3.1 further processing the medicinal materials weighed in step 2 to prepare the required ginseng-astragalus particle alcohol extract powder, wherein the main process comprises the following steps of:

firstly, the medicinal materials are crushed into coarse powder by using a powder grinding machine and fully and uniformly mixed, then the coarse powder is poured into a spherical flask containing 85vol% ethanol solution with the volume of 8 times of the medicinal powder, the spherical flask is heated to 80 ℃, the evaporated ethanol extract is recovered by a condensing tube, the heating lasts for 5 hours, after the medicinal solution is poured out, the ethanol heating and extracting process is repeated, the ethanol extract is recovered, the ethanol extracts of the two times are combined and placed in a water bath with the temperature of 50 ℃, the ethanol is recovered by a rotary evaporator, and the extracted liquid medicine is concentrated. And finally, freeze-drying the concentrated liquid medicine through a freeze dryer at the temperature of minus 56 ℃ to obtain extract powder, taking out the extract powder, grinding the extract powder into fine powder through a mortar, and filtering the fine powder through a 200-mesh filter screen to obtain the traditional Chinese medicine compound preparation (ginseng yellow particles).

4 mass spectrum analysis of the Chinese medicinal compound preparation (SHENHUANG granule) to explore its specific components

The method comprises the steps of carrying out mass spectrum data acquisition on ginseng yellow particles by using an ultra-high performance liquid chromatography-high resolution mass spectrometry (UPLC-Q-TOF-MS), analyzing chemical components in the ginseng yellow particles by combining literature reports and natural product high resolution mass spectrum database software 1.0 (Shanghai Shidande technology Co., ltd.), and identifying 45 compounds respectively: O-Galleylglucose, gallicic acid, phenprobamate, neachlorogenic acid, mesaconine, salidroside, catechin, chlorgenic acid, cryptochlorogenic acid, fuziline, eleutheroside E, chicorie acid, aloe-Emodin-8-O-beta-D-glucopyranoside, rhein-8-O-beta-D-glucopyranoside, benzoylmesaconine, emodin-1-O-glucoside, benzoylaconine, 3 (or 8) - (beta-D-glucopyranosidyl) -9,10-dihydro-8 (or 3) -hydroxy-1-methyl-9, 10-dihydroxy-2-anthracenecarboxylic add, benzoythypaconine, arctin ginsenosides Rg1, ginsenosides Re, hypaconitine, 3-Deokyhokbusine A, chrysopianol-8-0-. Beta. -D-glucopyranoside Aloe-Emodin-3- (hydroxyymethyl) -O-. Beta. -D-glucopyranoside, erodin-8-O-glucoside, pliyscion-O-. Beta. -D-monoglucoside, ginsenosides Rf, aloe-erodine, ginsenosides Rb1, ginsenosides Ro, ginsenosides Rb2, rhein, ginsenosides Rd, 2-Anthracenecarboxylie acid,9,10-dihydro-8-hydroxy-1-methyl-9, 10-diox-3- [ [6-O- [ (2E) -1-oxo-3-phenyl-2-propen-1yl ] - β -D-glucopyranosyl ] -, ginsenoside Rg4, ginsenoside Rh4, emodin, chikusetsusaponin Iva, ginsenoside Rg3 isomer, chrysophanol, ginsenoside Rg3 isomer, ginsenoside Rk1, cinsenoside Rg5.

Batch molecular docking of 45 compounds in the ginseng yellow particles with the TWIK2 target (fig. 2) was performed using SeeSAR 11.1.0 software, and 10 chemical components in the ginseng yellow particles were found: ginsenoside Re, arctiin, benzoylaconitine, benzoylmesaconine, aconitine, chicoric acid, phenylalanine, neochlorogenic acid and chlorogenic acid show good binding performance with TWIK 2.

However, the prior researches find that a key protein capable of transporting potassium ions to cross cell membranes to trigger inflammation is as follows: the double-pore-domain weak inward rectifying potassium channel 2 (Tandem ofpore domains in a weak inward rectifying K +channel 2, TWIK2) can regulate and control a potassium outflow-mediated signal transduction pathway, and plays an important role in the occurrence and development of sepsis ARDS. This example demonstrates that a variety of active ingredients in the ginseng yellow particles bind well to TWIK2 and reduce expression of the TWIK2 protein, revealing the potential of ginseng yellow particles to treat sepsis ARDS by modulating the TWIK2-NLRP3 signaling pathway.

EXAMPLE 2 therapeutic Effect of Shenhuang particles on sepsis ARDS mice

Early studies in the present invention demonstrated that a murine model of sepsis ARDS could be established by Lipopolysaccharide (LPS) induction, and that its lung tissue could exhibit significant alveolar hyperemia, exudation, interstitial and alveolar cell infiltration and epithelial cell damage, whereas the normal control mice had normal lung tissue structure (fig. 3).

1 preparation of experimental animal and model

Male wild type C57BL/6 mice, 20-25 g in mass, were provided by Shanghai university of traditional Chinese medicine laboratory animal center and randomly divided into control group, model group and Shenhuang granule (SHG) group. Dripping 50 mu LPBS solution into the airway of the control group; instilling 50 mu L of LPS (2 mg/kg) solution into the airway of the model group; SHG mice were first pre-treated with 50mg/kg SHG and after 2h, 50. Mu.L LPS solution was instilled into the airways. After 8h of LPS instillation, all mice were sacrificed and the following index tests were performed.

2 index detection

2.1 pathological detection

Mice were sacrificed, left lungs were isolated, fixed in 10% neutral formalin-fixed solution, and lung tissue paraffin sections were prepared and eosin-hematoxylin stained (HE stained).

2.2 arterial blood and pulmonary tissue detection index

PaO2/FiO2: the mice were anesthetized, cannulated with carotid artery occlusion, arterial blood samples were collected and analyzed for PaO2/FiO2 ratio.

Lung injury score: scoring the lung injury of the mice by a scoring system; the method comprises five parameters: exudation, neutrophil infiltration, alveolar hyperemia, protein fragments and alveolar septum thickening; the severity of each parameter was scored as follows: 0 = none; 1 = mild; 2 = medium; 3 = severe, average score per mouse was lung injury score.

After the mice are sacrificed, the two lungs are separated, rinsed, and immediately weighed on a balance after the sterile filter paper is wiped dry, and recorded as the wet weight of the lungs; the double lung tissue was then baked in an oven for one week and then re-weighed, recorded as the dry weight of the lung, and the wet/dry weight ratio of the lung was calculated.

Curative effect of 3 Shenhuang particles on sepsis ARDS mice

The lung tissue injury score of sepsis ARDS mice was reduced, paO2/FiO2 ratio was increased and lung wet/dry weight ratio was increased by ginseng yellow granule treatment (fig. 4).

EXAMPLE 3 clinical efficacy of Shenhuang particles on patients with sepsis ARDS

1 experiment design

(1) Inclusion criteria: (1) meets the main points of sepsis diagnosis in Chinese sepsis/septic shock emergency treatment guidelines (2018), and meets the main points of acute lung injury diagnosis in acute respiratory distress syndrome diagnosis and treatment guidelines (2006); (2) basic vital signs are basically stable; (3) acute physiological and chronic health scores (APACHE-II scores) are below 12 points; (4) the expected survival time is above 7 d.

(2) Exclusion criteria: (1) a variety of organ failure; (2) immunosuppressed persons are present; (3) for patients with serious allergic disease history in the past of the medicines used in the study; (4) those who had received other medications prior to participation in the study; (5) blurred consciousness or cognitive and mental disorders.

(3) And (3) drug intervention: patients received 14 days of treatment and underwent laboratory checks (whole blood count, general urine analysis, fecal occult blood test, liver function and kidney function) on days 1, 3, 5, 7 and 14 after the start of the test. The ginseng-astragalus particles are dissolved in warm water and orally taken, and the administration scheme is that each bag is provided with 100mL of dosage in two small bags each day. For critical patients who are difficult to take medicine, the full dose of the ginseng-astragalus granule solution is administered through a feeding tube. After 14 days, the ginseng-yellow granules were administered twice daily until death or discharge. According to the standard of the novel coronavirus infection diagnosis and treatment guide (ten edition of trial) in China, 2022. One group was given standard of care in combination with the ginseng yellow granules as SHG group ((30 patients), the other group was given standard of care only as control group ((30 patients).

(4) And (3) observing the indexes: overall improvement rate, overall mortality, rate of progression of disease, mechanical ventilation rate.

2 clinical efficacy

The overall disease improvement rate for patients with sepsis ARDS was 61.4% (35/57) with trepang-huang granule treatment, whereas the control group was 24.1% (13/54) (p < 0.01). Wherein, in the severe patients, the improvement rate of the control group is 41.2% (7/17), which is obviously lower than 94.7% (18/19) of the SHG group (p < 0.01), and in the critical patients, the improvement rate of the control group is 16.2% (6/37), which is lower than 44.7% (p < 0.01) of the SHG group (A in FIG. 5). After ginseng-astragalus particle treatment, the total mortality rate of the disease is 38.6%, the control group is 75.9%, and the total mortality rate is reduced by 37.3%. Wherein, in severe patients, the mortality rate of the control group is 58.8% which is significantly higher than that of the ginseng-yellow granule group by 5.3%, and in critical patients, the mortality rate of the control group is 83.8% which is higher than that of the ginseng-yellow granule group by 55.3% (B in FIG. 5).

In terms of disease progression (FIG. 6A), 88.2% (15/17) of the control group progressed to critically ill, significantly higher than 47.4% (9/19) of the ginseng-yellow granule group (p < 0.01); in terms of mechanical ventilation, the control group uses an invasive mechanical ventilation ratio of 58.8% (10/17), while the patient ratio of the ginseng-astragalus granule group is 0, and the clinical effect is obviously superior to that of pure western medicine treatment (p < 0.01).

Clinical experiments show that the ginseng-astragalus particles can obviously improve the related symptoms of sepsis ARDS in both in-vivo experiments and in-vitro experiments, can obviously reduce the death rate of patients, can effectively delay the progress of the illness state, and can greatly reduce the invasive mechanical ventilation proportion of the patients.

The above embodiments are only illustrative of the preferred embodiments of the present invention and are not intended to limit the scope of the present invention, and various modifications and improvements made by those skilled in the art to the technical solutions of the present invention should fall within the protection scope defined by the claims of the present invention without departing from the design spirit of the present invention.

Claims

1. A traditional Chinese medicine compound preparation for treating sepsis ARDS is characterized by comprising the following components in parts by weight:

2. The compound Chinese medicinal preparation according to claim 1, which is characterized by comprising the following components in parts by weight:

3. A method for preparing a compound Chinese medicinal preparation according to any one of claims 1-2, comprising the steps of:

4. The method according to claim 3, wherein the ethanol is an 85vol% ethanol solution, and the volume ratio of the medicinal material to the ethanol is 1: (8-10); the conditions of the ethanol heating reflux extraction are as follows: heating at 80deg.C, and reflux extracting for 2-3 times.

5. Use of a compound Chinese medicinal preparation according to claim 1 or 2 in the preparation of a medicament for the treatment of sepsis ARDS.

6. The use according to claim 5, wherein the compound Chinese medicinal preparation has the effect of treating sepsis ARDS by regulating and controlling the TWIK2-NLRP3 signaling pathway, improving symptoms of sepsis ARDS patients, reducing mortality and delaying disease progression.

7. The use according to claim 5, wherein the medicament further comprises pharmaceutically acceptable excipients.

8. The use according to claim 7, wherein the pharmaceutically acceptable excipients comprise any one or more of fillers, disintegrants, binders, lubricants and flavouring agents.