Traditional Chinese medicine composition for preventing or treating sepsis and target organ damage, preparation method and application

Abstract

The invention discloses a Chinese medicine composition for preventing or treating sepsis and target organ damage, a preparation method and an application thereof; the Chinese medicine composition comprises, by weight, 10-15 parts of rhubarb, 10-15 parts of immature bitter orange, 15-25 parts of magnolia bark, 12-23 parts of angelica, 38-52 parts of peony root, 10-18 parts of prepared rehmannia root, 32-48 parts of astragalus, 10-15 parts of white mulberry bark, 8-17 parts of raw liquorice, 15-25 parts of ophiopogon, 10-15 parts of aster and 9-16 parts of hovenia dulcis; the components of the Chinese medicine composition can coordinate and promote each other, play a role in reducing sepsis and target organ damage through synergistic enhancement, have definite efficacy in animal experiments, and can promote the standardized treatment of traditional Chinese medicine in the field of sepsis and target organ damage after being made into a standardized Chinese patent medicine preparation.

Description

Technical Field

[0001] The present invention belongs to the technical field of Chinese medicine compositions, and in particular relates to a Chinese medicine composition for preventing or treating sepsis and target organ damage, a preparation method and application thereof. Background Art

[0002] Sepsis is a life-threatening organ dysfunction caused by the body's dysregulated response to infection. It is one of the main causes of death in critically ill patients, with a mortality rate of up to 50%. Due to their physiological and structural characteristics, the lungs and liver are important target organs that are susceptible to damage, and the incidence of acute lung injury, acute respiratory distress syndrome, and acute liver failure is also the highest. So far, there is no specific treatment for sepsis and target organ damage.

[0003] In recent years, traditional Chinese medicine has been increasingly valued in the treatment of sepsis and target organ damage. There is no discussion or record of the corresponding disease names of sepsis and target organ damage in traditional Chinese medicine. According to its pathogenic characteristics, it belongs to the category of "toxic" pathogenicity in traditional Chinese medicine. Its basic etiology and pathogenesis are that toxic blood stasis cannot be transformed, and it is caused by the "Yangming Fu" - the large intestine: toxic evil accumulates in the intestine, the intestinal conduction is dysfunctional, and the water transportation is abnormal, resulting in constipation. The lung and the large intestine are exterior and interior, which leads to the failure of the lung to descend; the liver and the large intestine are connected, and the liver fails to dredge, which manifests as liver and lung damage related diseases. However, the current treatment effect for sepsis and target organ damage is not obvious. In view of the therapeutic advantages of traditional Chinese medicine in sepsis and target organ damage, it is of great significance to develop a Chinese medicine compound composition for the treatment of sepsis and target organ damage. Summary of the invention

[0004] The purpose of the present invention is to propose a Chinese medicine composition, a preparation method and an application for preventing or treating sepsis and target organ damage, so as to promote the standardized treatment of sepsis and target organ damage in the field of traditional Chinese medicine.

[0005] In view of this, the scheme of the present invention is as follows:

[0006] The first aspect of the present invention is to provide a Chinese medicine composition, which comprises the following components by weight: 10-15 parts of rhubarb, 10-15 parts of immature bitter orange, 15-25 parts of magnolia bark, 12-23 parts of angelica, 38-52 parts of peony root, 10-18 parts of rehmannia root, 32-48 parts of astragalus, 10-15 parts of white mulberry bark, 8-17 parts of raw liquorice, 15-25 parts of ophiopogon, 10-15 parts of aster, and 9-16 parts of Hovenia dulcis.

[0007] Furthermore, the composition comprises the following components by weight: 12-13 parts of rhubarb, 11-14 parts of immature bitter orange, 16-24 parts of magnolia bark, 15-20 parts of angelica, 40-50 parts of peony, 13-15 parts of rehmannia root, 35-45 parts of astragalus, 11-14 parts of mulberry bark, 10-15 parts of raw liquorice, 15-20 parts of ophiopogon, 11-14 parts of aster, and 10-15 parts of hovenia dulcis.

[0008] Furthermore, the composition comprises the following components by weight: 12 parts of rhubarb, 13 parts of immature bitter orange, 20 parts of magnolia bark, 18 parts of angelica, 45 parts of peony, 15 parts of rehmannia root, 40 parts of astragalus, 13 parts of mulberry bark, 12 parts of raw licorice, 20 parts of ophiopogon, 12 parts of aster, and 13 parts of hovenia dulcis.

[0009] The second aspect of the present invention is to provide a method for preparing a Chinese medicine preparation, using the Chinese medicine composition described in the first aspect, the steps comprising:

[0010] Mix Magnolia officinalis, Citrus aurantium and water, and boil them over high heat for 20 to 30 minutes to obtain liquid medicine 1;

[0011] Soak rhubarb, angelica, peony, rehmannia root, astragalus, white mulberry bark, raw licorice, ophiopogon, aster and Hovenia dulcis in water, add liquid one after soaking, then boil over high heat for 10 to 20 minutes, then simmer over low heat for 30 to 40 minutes, filter after boiling to obtain liquid two and filter residue; concentrate liquid two to make.

[0012] Furthermore, the preparation method also includes the step of leaching and recovering the filter residue: adding water to the filter residue, boiling over high heat for 10 to 20 minutes, decocting over low heat for 15 to 25 minutes, filtering after the decoction is completed to obtain medicinal solution three, and then combining medicinal solution two and medicinal solution three and concentrating them.

[0013] Furthermore, the mass volume ratio of the Chinese medicinal components to water in the medicinal liquid one is 1g: (10-15) mL; the mass volume ratio of the Chinese medicinal components to water in the medicinal liquid two is 1g: (10-15) mL; the mass volume ratio of the filter residue to water in the medicinal liquid three is 1g: (6-10) mL; and the concentration step concentrates the medicinal liquid to a relative density of 2.0-2.2 g / mL.

[0014] Preferably, the mass volume ratio of the Chinese medicinal components to water in the medicinal liquid one is 1g:12mL; the mass volume ratio of the Chinese medicinal components to water in the medicinal liquid two is 1g:12mL; the mass volume ratio of the filter residue to water in the medicinal liquid three is 1g:8mL; and the concentration step concentrates the medicinal liquid to a relative density of 2.1g / mL.

[0015] The third aspect of the present invention is to propose the use of the traditional Chinese medicine composition proposed in the first aspect in the preparation of drugs for preventing or treating sepsis and target organ damage.

[0016] Furthermore, the medicine also includes pharmaceutically acceptable excipients.

[0017] Furthermore, the medicine is an oral preparation. Preferably, the oral preparation includes an extract, granules, tablets, capsules or liquid preparations.

[0018] Compared with the prior art, the beneficial effects of the present invention include but are not limited to:

[0019] 1. The Chinese medicine composition provided by the present invention adopts a scientific ratio, can clear the bowels and detoxify, promote blood circulation and remove turbidity, strengthen the body and consolidate the foundation, and the various components can coordinate and promote each other, and play a role in alleviating sepsis and target organ damage through synergistic enhancement. Animal experiments have definite efficacy, and after being made into a standardized Chinese patent medicine preparation, it can promote the standardized treatment of traditional Chinese medicine in the field of sepsis and target organ damage.

[0020] 2. The Chinese medicine composition provided by the present invention is safe and reliable, has obvious clinical effects, simple preparation process, is suitable for industrial production, and enriches the treatment methods for sepsis and target organ damage. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] 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 creative work.

[0022] Figure 1 This is the TIC overlap spectrum of the metabolites of the traditional Chinese medicine compound in positive ion mode (Pos).

[0023] Figure 2 This is the TIC overlapping spectrum of the negative ion mode (Neg) of the metabolites of the traditional Chinese medicine compound.

[0024] Figure 3 The severity of the disease in each group of septic mice was scored.

[0025] Figure 4 The effect of a Chinese herbal compound on lung pathology in septic mice.

[0026] Figure 5 These are the results of the effect of the Chinese herbal compound on liver pathology in septic mice.

[0027] Figure 6 A is the effect of the Chinese medicine compound on intestinal histopathology in septic mice; Figure 6 B shows the grading of intestinal mucosal pathological damage in mice in each group.

[0028] Figure 7 The results show the effect of the Chinese herbal compound on the wet / dry weight ratio of the lungs of septic mice.

[0029] Figure 8 These are the results of the effect of the Chinese herbal compound on inflammatory factors in mice. DETAILED DESCRIPTION

[0030] The following provides definitions of some terms used in this specification. Unless otherwise specified, all terms used herein are generally understood by those skilled in the art to which this solution belongs.

[0031] In one embodiment, a Chinese medicine composition is proposed, wherein the components by weight are: 10-15 parts of rhubarb, 10-15 parts of immature bitter orange, 15-25 parts of magnolia bark, 12-23 parts of angelica, 38-52 parts of peony, 10-18 parts of rehmannia root, 32-48 parts of astragalus, 10-15 parts of mulberry bark, 8-17 parts of raw licorice, 15-25 parts of ophiopogon, 10-15 parts of aster, and 9-16 parts of Hovenia dulcis.

[0032] In the above embodiments, the Chinese medicine composition adopts a scientific ratio, which can reduce the damage to the lungs, liver and intestines related to sepsis. The components can coordinate and promote each other, and play a synergistic role. Traditional Chinese medicine believes that in sepsis, "turbid toxins penetrate the internal organs" presents the characteristics of rapid onset, rapid changes, toxic evils often with phlegm and blood stasis, and lingering and difficult to cure. External and endogenous turbid toxins disturb the internal organs by penetrating the Yingwei, circulating the meridians, and infiltrating the qi and blood. Traditional Chinese medicine treatment should take "clearing the bowels and removing turbidity, detoxifying and protecting the internal organs" as the general Chinese medicine treatment method. The Chinese medicine compound composition of the present invention can clear the bowels and detoxify, activate blood circulation and remove turbidity, strengthen the body, and play a role in reducing sepsis and target organ damage. Animal experiments have definite efficacy. After being made into standardized Chinese patent medicine preparations, it can promote the standardized treatment of traditional Chinese medicine in the field of sepsis and target organ damage.

[0033] In the above embodiments, the Chinese medicine composition is safe and reliable, and has obvious clinical effects. It can reduce the levels of multiple inflammatory mediators such as IL-1β, IL-6, TNF-α in the mouse sepsis model, reduce the systemic inflammatory response of the septic mice, and improve the prognosis. The Chinese medicine composition can reduce the pathological damage of the lung, liver and intestinal tissues of the sepsis model mice, reduce the wet / dry weight ratio of the lungs, reduce the levels of liver function indicators ALT and AST, improve the Chiu score of intestinal mucosal injury, reduce the intestinal permeability of mice, protect the organ function, and prevent the occurrence of target organ function damage.

[0034] In another embodiment, a method for preparing a Chinese medicine preparation is provided, using the Chinese medicine composition described in the above embodiment, the steps comprising:

[0035] Mix Magnolia officinalis, Citrus aurantium and water, and boil them over high heat for 20 to 30 minutes to obtain liquid medicine 1;

[0036] Soak rhubarb, angelica, peony, rehmannia root, astragalus, white mulberry bark, raw licorice, ophiopogon, aster and Hovenia dulcis in water, add liquid one after soaking, then boil over high heat for 10 to 20 minutes, then simmer over low heat for 30 to 40 minutes, filter after boiling to obtain liquid two and filter residue; concentrate liquid two to make.

[0037] Furthermore, the preparation method also includes the step of leaching and recovering the filter residue: adding water to the filter residue, boiling over high heat for 10 to 20 minutes, decocting over low heat for 15 to 25 minutes, filtering after the decoction is completed to obtain medicinal solution three, and then combining medicinal solution two and medicinal solution three and concentrating them.

[0038] Furthermore, the mass volume ratio of the Chinese medicinal components to water in the medicinal liquid one is 1g: (10-15) mL; the mass volume ratio of the Chinese medicinal components to water in the medicinal liquid two is 1g: (10-15) mL; the mass volume ratio of the filter residue to water in the medicinal liquid three is 1g: (6-10) mL; and the concentration step concentrates the medicinal liquid to a relative density of 2.0-2.2 g / mL.

[0039] Preferably, the mass volume ratio of the Chinese medicinal components to water in the medicinal liquid one is 1g:12mL; the mass volume ratio of the Chinese medicinal components to water in the medicinal liquid two is 1g:12mL; the mass volume ratio of the filter residue to water in the medicinal liquid three is 1g:8mL; and the concentration step concentrates the medicinal liquid to a relative density of 2.1g / mL.

[0040] In the above embodiments, the preparation process is simple, suitable for industrial production, and enriches the treatment methods for sepsis and target organ damage.

[0041] In an embodiment of the present invention, the pathological model of sepsis in mice was established by cecal ligation and perforation. The specific modeling method is as follows: 1.5% sodium pentobarbital is taken at a dose of 30 mg / kg for intraperitoneal injection anesthesia. After disinfection, a 1.5 cm long incision is made along the midline of the abdomen, and then the cecum is gently separated, and the cecum is ligated in a circular manner with a 4-0 silk thread. After puncturing the free end with a 19G needle once, a small amount of feces is squeezed out, the cecum is restored, and finally the abdominal cavity is sutured and closed. The mice in the sham operation group only received the operation of opening the abdomen, turning the cecum over, restoring it, and closing the abdomen. The mice were immediately subcutaneously injected with 2 ml of normal saline for fluid replacement after surgery. The mice were killed and the samples were collected 24 hours later.

[0042] Example 1

[0043] A traditional Chinese medicine composition for treating sepsis and target organ damage comprises the following components by weight: 12 parts of rhubarb, 13 parts of immature bitter orange, 20 parts of magnolia bark, 18 parts of angelica, 45 parts of peony root, 15 parts of prepared rehmannia root, 40 parts of astragalus, 13 parts of white mulberry bark, 12 parts of raw liquorice, 20 parts of ophiopogon, 12 parts of aster and 13 parts of hovenia dulcis.

[0044] Preparation method:

[0045] 1) Mix Magnolia officinalis, Citrus aurantium and water, with the total mass of Magnolia officinalis and Citrus aurantium to the volume ratio of water being 1 g:12 mL, and first boil over high heat for 30 minutes to obtain liquid 1;

[0046] 2) Add water to rhubarb, angelica, peony, prepared rehmannia root, astragalus, white mulberry bark, raw licorice, ophiopogon, aster and Hovenia dulcis; the volume ratio of the total weight of rhubarb, angelica, peony, prepared rehmannia root, astragalus, white mulberry bark, raw licorice, ophiopogon, aster and Hovenia dulcis to water is 1g:12mL. Soak for 30 minutes, add liquid 1 after soaking, then boil for 30 minutes over high heat, then decoct for 30 minutes over low heat, filter with 8 layers of gauze after decoction to obtain liquid 2 and filter residue;

[0047] 3) Add water to the filter residue, the mass volume ratio of the filter residue to water is 1g:8mL, boil over high heat for 20 minutes, then simmer over low heat for 20 minutes, filter with 8 layers of gauze after decocting to obtain liquid three;

[0048] 4) Combine the second and third medicinal solutions, and concentrate under reduced pressure at 40° C. to 1 / 4 of the original volume to obtain a concentrated solution with a relative density of 2.10 g / mL, i.e., a Chinese medicine composition preparation; store in a refrigerator at 4° C. for later use.

[0049] Example 2

[0050] A traditional Chinese medicine composition for treating sepsis and target organ damage comprises the following components by weight: 10 parts of rhubarb, 15 parts of immature bitter orange, 15 parts of magnolia bark, 23 parts of angelica, 38 parts of peony, 18 parts of rehmannia root, 32 parts of astragalus, 15 parts of white mulberry bark, 8 parts of raw liquorice, 25 parts of ophiopogon, 10 parts of aster and 16 parts of hovenia dulcis.

[0051] Preparation method:

[0052] 1) Mix Magnolia officinalis, Citrus aurantium and water, with the total mass of Magnolia officinalis and Citrus aurantium to the volume ratio of water being 1 g:10 mL, and first boil over high heat for 30 minutes to obtain liquid 1;

[0053] 2) Add water to rhubarb, angelica, peony, prepared rehmannia root, astragalus, white mulberry bark, raw licorice, ophiopogon, aster and Hovenia dulcis; the volume ratio of the total weight of rhubarb, angelica, peony, prepared rehmannia root, astragalus, white mulberry bark, raw licorice, ophiopogon, aster and Hovenia dulcis to water is 1g:15mL. Soak for 30 minutes, add liquid 1 after soaking, then boil for 30 minutes over high heat, then decoct for 30 minutes over low heat, filter with 8 layers of gauze after decoction to obtain liquid 2 and filter residue;

[0054] 3) Add water to the filter residue, the mass volume ratio of the filter residue to water is 1g:10mL, boil over high heat for 20 minutes, then simmer over low heat for 20 minutes, filter with 8 layers of gauze after decocting to obtain liquid three;

[0055] 4) Combine the second and third medicinal solutions, and concentrate under reduced pressure at 40° C. to 1 / 6 of the original volume to obtain a concentrated solution with a relative density of 2.14 g / mL, i.e., a Chinese medicine composition preparation; store in a refrigerator at 4° C. for later use.

[0056] Example 3

[0057] A traditional Chinese medicine composition for treating sepsis and target organ damage comprises the following components by weight: 15 parts of rhubarb, 10 parts of immature bitter orange, 25 parts of magnolia bark, 12 parts of angelica, 52 parts of peony root, 10 parts of prepared rehmannia root, 45 parts of astragalus, 11 parts of white mulberry bark, 17 parts of raw liquorice, 15 parts of ophiopogon, 15 parts of aster and 9 parts of hovenia dulcis.

[0058] Preparation method:

[0059] 1) Mix Magnolia officinalis, Citrus aurantium and water, with the total mass of Magnolia officinalis and Citrus aurantium to the volume ratio of water being 1 g:15 mL, and boil over high heat for 30 minutes to obtain liquid 1;

[0060] 2) Add water to rhubarb, angelica, peony, prepared rehmannia root, astragalus, white mulberry bark, raw licorice, ophiopogon, aster and Hovenia dulcis; the volume ratio of the total weight of rhubarb, angelica, peony, prepared rehmannia root, astragalus, white mulberry bark, raw licorice, ophiopogon, aster and Hovenia dulcis to water is 1g:10mL. Soak for 30 minutes, add liquid 1 after soaking, then boil for 30 minutes over high heat, then decoct for 30 minutes over low heat, filter with 8 layers of gauze after decoction to obtain liquid 2 and filter residue;

[0061] 3) Add water to the filter residue, the mass volume ratio of the filter residue to water is 1g:6mL, boil over high heat for 20 minutes, then simmer over low heat for 20 minutes, filter with 8 layers of gauze after decocting to obtain liquid three;

[0062] 4) Combine the second and third medicinal solutions, and concentrate under reduced pressure at 40° C. to 1 / 4 of the original volume to obtain a concentrated solution with a relative density of 2.11 g / mL, i.e., a Chinese medicine composition preparation; store in a refrigerator at 4° C. for later use.

[0063] Comparative Example 1

[0064] Provided is a Chinese medicine composition, which differs from Example 1 in that Hovenia dulcis is omitted, and the remaining components are the same as those in Example 1, and the preparation method is also the same as that in Example 1.

[0065] Comparative Example 2

[0066] Provided is a Chinese medicine composition, which differs from Example 1 in that cooked rehmannia root is replaced with raw rehmannia root, and the remaining components are the same as those in Example 1, and the preparation method is also the same as that in Example 1.

[0067] Comparative Example 3

[0068] Provided is a Chinese medicine composition, which differs from Example 1 in that peony is omitted, and the remaining components are the same as those in Example 1, and the preparation method is also the same as that in Example 1.

[0069] Comparative Example 4

[0070] Provided is a Chinese medicine composition, which differs from Example 1 in that Radix Ophiopogonis is omitted, and the remaining components are the same as those in Example 1, and the preparation method is also the same as that in Example 1.

[0071] Test Example 1

[0072] 56 mice were divided into a blank control group, a sham operation group, and a model group. The model group was further randomly divided into 5 groups, each with 8 mice. Each group was fed with the Chinese medicine composition in Example 1 of the present invention and Comparative Examples 1 to 4, twice a day, 8 mL each time. After the administration, the carotid artery blood of each group of mice was collected to detect the expression levels of inflammatory factors IL-1β, IL-6 and TNF-α. The results are shown in Table 1.

[0073] Table 1:

[0074] Grouping IL-1β (pg / mL) IL-6 (pg / mL) TNF-α(pg / mL) Sham operation group 89.46±13.97 65.70±13.10 87.88±8.91 Model Group 474.34±19.20 680.62±24.12 563.71±45.31 Example 1 140.45±13.37 115.38±66.88 183.47±12.80 Example 2 149.36±14.12 129.21±35.73 192.60±45.52 Example 3 148.71±13.58 127.54±49.68 190.12±42.43 Comparative Example 1 357.22±19.32 215.77±22.63 395.97±28.54 Comparative Example 2 297.60±21.57 415.98±12.24 334.56±31.23 Comparative Example 3 322.30±12.29 364.54±24.34 290.73±23.15 Comparative Example 4 280.99±22.54 478.79±29.74 465.96±42.18

[0075] As can be seen from the above table, compared with the sham operation group, the expression levels of IL-1β, IL-6 and TNF-α in the model group were significantly increased, indicating that the modeling of the present invention was successful. Compared with comparative examples 1 to 4, after taking the Chinese medicine composition of Examples 1 to 3 of the present invention, the expression levels of IL-1β, IL-6 and TNF-α in mice can be significantly reduced. This shows that the Chinese medicine composition of the present invention can effectively eliminate the inflammatory response caused by sepsis. And Example 1 has the best effect on reducing the expression levels of IL-1β, IL-6 and TNF-α in mice relative to Examples 2-3, and can be used as the best example. After the components of Comparative Examples 1 to 4 were reduced or replaced, the effect of eliminating the inflammatory response caused by sepsis was weakened, indicating that there is a synergistic effect between the components.

[0076] Test Example 2 Detection of main metabolic components in traditional Chinese medicine composition

[0077] Ultra-high performance liquid chromatography-mass spectrometry (UHPLC-Q-TOF MS) was used to detect the types and contents of ingredients in the traditional Chinese medicine composition.

[0078] Weigh 200 μl of the liquid preparation of the Chinese medicine compound composition prepared in Example 1, add 80% methanol to 1 ml, vortex for 1 min, centrifuge at 14000 rpm for 10 min, start sampling, and use UHPLC-Q-TOF MS technology to perform qualitative and quantitative analysis of metabolites in the Chinese medicine compound composition. The chromatograms of each analyte were collected and integrated by Xcilabur4.1 software (Thermo Fisher), and the 1 / X 2 Perform linear regression for the weighting coefficients.

[0079] UHPLC-Q-TOF MS technology was used to conduct qualitative and quantitative analysis of metabolites in traditional Chinese medicine compound. The TIC overlap spectrum of metabolites in positive ion mode (Pos) and negative ion mode (Neg) are shown in Figure 2. Figure 1 , Figure 2 As shown. The positive and negative ion chromatograms (TIC) of the samples were overlapped and compared, and the results showed that the response intensity and retention time of each chromatographic peak overlapped, indicating that the changes caused by instrument errors were very small during the entire experiment. By matching the molecular mass, secondary fragmentation spectrum, retention time and other information of the metabolites contained in the Chinese Science New Life Plant Metabolome Database, the metabolites in the Chinese herbal compound composition samples were structurally identified and verified, and the identification level was determined to be at level 1 and level 2. The total metabolites were identified by combining positive and negative ion modes. The number of metabolites identified in the positive ion mode (Pos) was 439, and the number of metabolites identified in the negative ion mode (Neg) was 319. Among them, the main metabolic components of the Chinese herbal compound composition include flavonoids, organic oxygen compounds, benzene and its substituted derivatives, fatty acyl groups, coumarins and their derivatives, arbutins, amyl alcohol lipids, phenols, protoberberine alkaloids and their derivatives, cinnamic acid and its derivatives, etc.

[0080] Test Example 3

[0081] 1. Breeding and modeling of sepsis mice

[0082] (1) Housing conditions of septic mice: 40 8-week-old Balb / c male mice (weight 18-22 g) were purchased from Beijing Biotechnology Co., Ltd. All mice were raised in the animal room of the Animal Experiment Center of Huazhong University of Science and Technology for animal experiments. The breeding environment was maintained at a constant temperature (24±2°C) and humidity (50±5%), and mice were allowed free access to food and water. At the same time, the animal room implemented a cyclic lighting system of 12 h light and 12 h dark to simulate the natural circadian rhythm. The experiment began after 1 week of adaptive feeding.

[0083] (2) Animal grouping, modeling and drug administration: 40 Balb / c mice were randomly divided into a sham operation group, a model group, and a low-, medium- and high-dose group of the Chinese medicine compound composition prepared in Example 1, with 8 mice in each group. After CLP modeling of mice, the five groups were given normal drinking water, and the low-, medium- and high-dose groups of the Chinese medicine compound composition were given intragastric administration of the Chinese medicine compound composition: the concentration of the low-dose group of the Chinese medicine compound composition was 2.5 g / kg, the concentration of the medium-dose group was 5 g / kg, and the concentration of the high-dose group was 10 g / kg. The sham operation group and the model group were given intragastric administration of equal volumes of distilled water, 12 hours before surgery and 12 hours after surgery, respectively, for a total of 2 times. The dosage calculation method was converted with reference to Chen Qi's "Methodology of Chinese Medicine Pharmacology Research".

[0084] 2. Observation indicators of mice

[0085] (1) Severity score of sepsis in mice: During modeling and drug administration, mice were scored using the MSS scoring system developed by Shrum et al., with the main observations being: appearance, consciousness, level of action, response to external stimuli, eye secretions, and respiratory quality. Each of the above items was scored between 0 and 4, with higher scores indicating a higher degree of sepsis severity.

[0086] (2) Histopathological observation of target organs (lung, liver, and intestine) of mice: After the experiment, the mice were killed and the lung, liver, and intestinal tissues were taken. After being rinsed with saline, they were placed in tissue fixative, embedded in paraffin, and sliced. The histopathological changes were observed after hematoxylin-eosin (HE) staining. The injury score was independently evaluated by two pathologists without knowing the sample grouping.

[0087] (3) Determination of wet / dry weight ratio of mouse lungs: When sampling, the right upper lung of the mouse was cut off and weighed immediately, then placed in an oven to dry for 72 h (60°C) and weighed again, and finally the wet / dry weight ratio of the lungs was calculated.

[0088] (4) The test kit was used to detect the changes in the levels of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) of liver function in each group of mice. The operation was carried out strictly according to the instructions of the instruction manual.

[0089] (5) Grading of intestinal mucosal pathological damage: The Chiu grading system is used to evaluate intestinal mucosal damage. Based on the status of the mucosal villi and glands, the intestinal mucosa is graded to see whether there are changes in the subcutaneous space, capillary congestion, lamina propria shedding, ulcers or bleeding. There are 5 grades in total, and the higher the grade, the more severe the damage.

[0090] (6) Changes in systemic inflammation levels in mice: ELISA was used to determine the levels of IL-6 and TNF-α in mouse serum: The ELISA method was used to determine the levels of IL-1β, IL-6, and TNF-α in mouse serum. The specific operation steps were strictly carried out according to the instructions of the kit.

[0091] 3. Experimental Results

[0092] (1) The Chinese herbal medicine compound can improve the severity of sepsis in mice

[0093] Figure 3 The results of the disease severity scores of septic mice in each group showed that 12 hours after cecal ligation and puncture, mice began to show some disease symptoms. 24 hours after surgery, the sepsis score of mice in the model group was significantly higher than that in the normal group. The mice showed mental depression, slow movement, reduced food intake, erect hair on the back, viscous secretions in the eyelids, turbid exudate in the abdominal cavity, intestinal adhesions, and odor, indicating that the mouse model was successfully established. Compared with the model group mice, the low, medium, and high dose groups of the Chinese herbal compound composition significantly reduced the sepsis score, and the scores of the three groups were 17.6 (P < 0.05), 11.5 (P < 0.01), and 6.3 (P < 0.01), respectively. The disease symptoms of mice were alleviated, which shows that the Chinese herbal compound composition can reduce the severity of the disease in septic mice.

[0094] (2) Chinese herbal medicines have protective effects on internal organs

[0095] Figure 4 The effect of the Chinese medicine compound on the lung pathology of septic mice was shown in Figure 2. Figure 4 It can be seen that compared with the sham operation group, the lung tissue of the mice in the model group showed obvious pathological damage changes. The HE staining results showed that there were a large number of inflammatory cell infiltration and hemorrhage in the alveolar cavity, lung tissue structure destruction, alveolar wall thickening and other changes. Compared with the model group, the degree of lung tissue damage in the low, medium and high dose groups of the Chinese medicine compound composition was reduced in a concentration-dependent manner. HE staining showed that alveolar wall thickening, lung tissue structure destruction, and lung inflammatory cell infiltration were significantly alleviated.

[0096] Figure 5 The effect of the Chinese medicine compound on liver pathology in septic mice was shown in Figure 2. Figure 5It can be seen that compared with the sham operation group, the liver cells of the mice in the model group were severely edematous, the cytoplasm was loose, the central vein hepatic sinusoids were dilated and congested, red blood cells were seen in the hepatic sinusoids, diffusely distributed spot and sheet necrosis appeared in the liver, diffuse inflammatory cells, lymphocyte infiltration, eosinophilic changes, some lobular structures were destroyed, some cell nuclei were fragmented, and the cytoplasm was darkly stained. The liver damage in the low, medium and high dose groups of the Chinese medicine compound composition was significantly alleviated compared with the model group, and the inflammatory cell infiltration was significantly reduced.

[0097] Figure 6 A is the effect of the Chinese medicine compound on intestinal tissue pathology in septic mice, Figure 6 B shows the grading of intestinal mucosal pathological damage in each group of mice. Figure 6 It can be seen that compared with the sham operation group, the intestinal mucosa of the model group mice showed disordered tight junction structure, reduced goblet cells and glands and disordered arrangement. The Chiu score of the model group was significantly higher than that of the control group (P < 0.01). Compared with the model group, the scores of the low, medium and high dose groups of the Chinese medicine compound composition gradually decreased (P < 0.05), the intestinal mucosal barrier was relatively intact, and the glands were neatly arranged. This shows that the Chinese medicine compound composition can alleviate intestinal barrier damage in sepsis.

[0098] Table 2 shows the effect of the Chinese herbal compound composition on the liver function of septic mice. It can be seen from Table 2 that compared with the sham operation group, the ALT and AST levels in the model group were significantly increased (P<0.01), while compared with the model group, the serum ALT and AST levels of mice were significantly decreased after administration of the Chinese herbal compound composition (P<0.01), and the higher the concentration, the more obvious the effect.

[0099] Table 2:

[0100] Grouping Dosage (g / kg) ALT / (U / L) AST / (U / L) Sham operation group - 49.65±3.46 74.06±2.45 Model Group - <![CDATA[274.34±13.78 ## ]]> <![CDATA[380.06±24.91 ## ]]> Low dose group 2.5 240.59±13.66* 322.36±23.1* Medium dose group 5 157.73±14.85* 215.29±22.65* High dose group 10 97.73±13.77** 115.29±12.53**

[0101] #P<0.05, ##P<0.01, relative to the model group, *P<0.05, **P<0.01.

[0102] Figure 7 The effect of the Chinese herbal medicine compound on the lung wet / dry weight ratio of septic mice was investigated. Figure 7 It can be seen that compared with the sham operation group, the wet / dry weight ratio of the lungs of the mice in the model group was significantly increased (P<0.01), while compared with the model group, the wet / dry weight ratio of the lungs of the low, medium and high dose groups of the Chinese medicine compound composition gradually decreased (P<0.05).

[0103] Figure 8 The effect of the Chinese medicine compound on inflammatory factors in mice. Figure 8It can be seen that compared with the sham operation group, the levels of IL-1β, IL-6, and TNF-α in the serum of the model group mice increased significantly (P<0.01); compared with the model group, the Chinese medicine compound group can significantly inhibit the levels of proinflammatory cytokines IL-1β, IL-6, and TNF-α in the serum of mice (P<0.01). The higher the concentration of the Chinese medicine compound, the more obvious the anti-inflammatory effect.

[0104] Although the present invention is disclosed as above, the protection scope of the present invention is not limited thereto. Those skilled in the art may make various changes and modifications without departing from the spirit and scope of the present invention, and these changes and modifications will fall within the protection scope of the present invention.

Claims

1. A Chinese medicine composition, characterized in that: The components are as follows by weight: 10-15 parts of rhubarb, 10-15 parts of immature bitter orange, 15-25 parts of magnolia bark, 12-23 parts of angelica, 38-52 parts of peony root, 10-18 parts of rehmannia root, 32-48 parts of astragalus, 10-15 parts of mulberry bark, 8-17 parts of raw liquorice, 15-25 parts of ophiopogon, 10-15 parts of aster, and 9-16 parts of Hovenia dulcis.

2. The Chinese medicine composition according to claim 1, characterized in that: The components are as follows by weight: 12-13 parts of rhubarb, 11-14 parts of immature bitter orange, 16-24 parts of magnolia bark, 15-20 parts of angelica, 40-50 parts of peony, 13-15 parts of rehmannia root, 35-45 parts of astragalus, 11-14 parts of mulberry bark, 10-15 parts of raw liquorice, 15-20 parts of ophiopogon, 11-14 parts of aster, and 10-15 parts of Hovenia dulcis.

3. The Chinese medicine composition according to claim 1, characterized in that The components are as follows by weight: 12 parts of rhubarb, 13 parts of immature bitter orange, 20 parts of magnolia bark, 18 parts of angelica, 45 parts of peony root, 15 parts of rehmannia root, 40 parts of astragalus, 13 parts of white mulberry bark, 12 parts of raw liquorice, 20 parts of ophiopogon, 12 parts of aster, and 13 parts of hovenia dulcis.

4. A method for preparing a Chinese medicine preparation, using the Chinese medicine composition according to any one of claims 1 to 3, characterized in that the steps include: Mix Magnolia officinalis, Citrus aurantium and water, and boil them over high heat for 20 to 30 minutes to obtain liquid medicine 1; Soak rhubarb, angelica, peony, rehmannia root, astragalus, white mulberry bark, raw licorice, ophiopogon, aster and Hovenia dulcis in water, add liquid one after soaking, then boil over high heat for 10 to 20 minutes, then simmer over low heat for 30 to 40 minutes, filter after boiling to obtain liquid two and filter residue; concentrate liquid two to make.

5. The preparation method according to claim 4, characterized in that: The method also includes the step of leaching and recovering the filter residue: adding water to the filter residue, boiling it over high heat for 10 to 20 minutes, decocting it over low heat for 15 to 25 minutes, filtering it after the decocting to obtain the third medicinal solution, and then combining the second medicinal solution and the third medicinal solution and concentrating them.

6. Use of the Chinese medicine composition according to any one of claims 1 to 3 in the preparation of a drug for preventing or treating target organ damage caused by sepsis, wherein the target organ is the lung, liver or intestine.

7. The use according to claim 6, characterized in that: The drug also includes pharmaceutically acceptable excipients.

8. The use according to claim 6, characterized in that: The medicine is an oral preparation.

9. The use according to claim 8, characterized in that: The oral preparations include extracts, granules, tablets, capsules or liquid preparations.

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