A pharmaceutical composition for treating sepsis, its preparation method and application

Abstract

This invention relates to the field of pharmaceutical composition technology, and more particularly to a pharmaceutical composition for treating sepsis and its preparation method. A pharmaceutical composition for treating sepsis is prepared from the following components in parts by weight: 20-26 parts of Andrographis paniculata, 18-24 parts of Forsythia suspensa, 17-21 parts of Anemarrhena asphodeloides, 15-19 parts of Vaccaria segetalis, 14-18 parts of Achyranthes bidentata, 12-16 parts of Scrophularia ningpoensis, 10-14 parts of Coptis chinensis, 7-11 parts of Melia toosendan, 5-9 parts of Prunus persica leaf, and 3-7 parts of Trillium tsao-ko. The formulation of this invention can reduce the inflammatory response of sepsis, alleviate apoptosis of intestinal mucosal epithelial cells in sepsis, promote cell survival, and thereby inhibit organ damage following sepsis.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical composition technology, and in particular to a pharmaceutical composition for treating sepsis, its preparation method, and its application. Background Technology

[0002] Sepsis is a systemic inflammatory response to infection and a common complication of severe infection, severe trauma, major surgery, severe acute pancreatitis, and shock. It can progress to septic shock and multiple organ dysfunction syndrome. Despite significant advancements in anti-infective therapy, surgical techniques, and critical care monitoring and treatment, the mortality rate of sepsis remains as high as 29%–38%. Improving the prognosis of sepsis patients is a major challenge and a critical issue in the field of emergency and critical care medicine.

[0003] Sepsis is a life-threatening organ dysfunction caused by a dysregulated response to infection. Therefore, developing targeted therapies to prevent or limit the progression of fatal organ failure, based on the pathogenesis of sepsis and the molecular mechanisms of its resulting organ failure, is an urgent treatment strategy. Professor Yao Yongming proposed immunomodulatory therapy for sepsis. In the later stages of sepsis, immune depletion occurs, and preventing "secondary infection" by regulating both the innate and adaptive immune systems is crucial for the prognosis of sepsis patients and the treatment of severe sepsis. However, since early sepsis damage is caused by a storm of excessively released cytokines leading to multi-organ damage, intervention targeting the early excessive inflammatory response is also a means of sepsis prevention and treatment.

[0004] Recent studies have recognized cell death as a crucial pathological change in sepsis. Previous research primarily focused on cell necrosis and apoptosis, neglecting a third mode of cell death—autophagy. Only recently has autophagy, a significant component of programmed cell death, gained increasing attention. Autophagy, meaning self-phagocytosis, possesses high species homology and is widely present in eukaryotic cells, playing roles in degrading faulty proteins, phagocytizing bacteria, and antigen presentation. In sepsis, autophagy can control the release of pro-inflammatory factors, thus providing a protective effect. Therefore, autophagy can be a research target, and its regulation could open new avenues for sepsis treatment. Studies have confirmed that in sepsis models, LPS stimulation and bacterial infection increase autophagy, which can protect cells and reduce cell death.

[0005] Among the many target organs damaged in sepsis, the gastrointestinal tract is the most vulnerable target organ when sepsis induces multiple organ dysfunction. At the same time, gastrointestinal dysfunction also plays an important role in the pathogenesis of sepsis and is often considered to be the initiating factor of sepsis-induced multiple organ failure and the initiating organ of sepsis.

[0006] The key to treating gastrointestinal dysfunction is maintaining the intestinal mucosal barrier, restoring intestinal continuity, and adjusting homeostasis, circulation, and oxygen supply. Sepsis most commonly affects the intestines, causing edema of the intestinal mucosal epithelium, rupture of epithelial cell membranes and intercellular junctions, cell necrosis, and epithelial shedding starting from the villus tips, sometimes even resulting in the complete shedding of the mucosal thickness and the formation of ulcers. Increased intestinal permeability, impaired mechanical barrier function, and translocation of bacteria and endotoxins exacerbate and destabilize SIRS, inducing MODS and endangering the patient's life. Therefore, the mechanical barrier formed by intestinal mucosal epithelial cells is the first line of defense against microbial invasion and the anatomical basis for the successful functioning of other barrier functions.

[0007] The pathogenesis of intestinal mucosal epithelial cell damage is closely related to endotoxemia. Endotoxemia can damage intestinal mucosal epithelial cells and disrupt barrier function through hypoxia, release of inflammatory mediators, and increased free radicals, leading to the translocation of a large number of bacteria and ultimately causing multiple organ failure and even death.

[0008] Therefore, protecting intestinal mucosal epithelial cells and barrier function, and alleviating gastrointestinal dysfunction are key issues for improving the prognosis of sepsis. Regulating the inflammatory response and reducing the sensitivity of intestinal mucosal epithelial cells to bacterial endotoxins is an important strategy. Summary of the Invention

[0009] The purpose of this invention is to provide a pharmaceutical composition for treating sepsis and a method for preparing the same.

[0010] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0011] This invention provides a pharmaceutical composition for treating sepsis, prepared from the following components in parts by weight: 20-26 parts of Andrographis paniculata, 18-24 parts of Forsythia suspensa, 17-21 parts of Anemarrhena asphodeloides, 15-19 parts of Vaccaria segetalis, 14-18 parts of Achyranthes bidentata, 12-16 parts of Scrophularia ningpoensis, 10-14 parts of Coptis chinensis, 7-11 parts of Melia toosendan, 5-9 parts of Prunus persica leaf, and 3-7 parts of Trillium truncatum.

[0012] The present invention also provides a method for preparing the pharmaceutical composition for treating sepsis, comprising the following steps:

[0013] (1) Mix Andrographis paniculata, Forsythia suspensa, Anemarrhena asphodeloides, Vaccaria segetalis, Achyranthes bidentata, Scrophularia ningpoensis, Coptis chinensis, Melia toosendan, Prunus persica leaves and Trillium truncata to obtain material 1;

[0014] (2) Mix the material 1, mixed enzyme and water for enzymatic hydrolysis, inactivate the enzyme, filter, centrifuge and take the supernatant;

[0015] (3) The supernatant is concentrated and dried to obtain a pharmaceutical composition for treating sepsis.

[0016] Preferably, the mass ratio of material 1, mixed enzyme and water in step (2) is 8-12:0.3-0.5:100.

[0017] Preferably, the mixed enzyme is cellulase, protease and α-amylase; the mass ratio of cellulase, protease and α-amylase is 12-16:5-9:7-11.

[0018] Preferably, the enzyme activities of the cellulase, protease, and α-amylase are all 3200–3800 U / g.

[0019] Preferably, the enzymatic hydrolysis in step (2) is carried out at a temperature of 42–48°C for 2.5–3.5 h.

[0020] Preferably, the centrifugation speed in step (2) is 2200-2800 rpm and the time is 5-9 min.

[0021] Preferably, the supernatant in step (3) is concentrated to 1 / 5 to 1 / 3 of its original volume.

[0022] Preferably, the drying in step (3) is spray drying; the inlet air temperature of the spray drying is 128-136°C and the outlet air temperature is 67-73°C.

[0023] The use of the above-described pharmaceutical composition or the pharmaceutical composition prepared by the above-described method in the preparation of a drug for treating sepsis.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] This invention establishes a rat model of sepsis using cecal ligation to investigate whether a prepared composition can alleviate intestinal mucosal epithelial cell damage in sepsis-affected rats by inducing autophagy. After cecal ligation, the serum levels of D-lactic acid, DAO, endotoxin, IL-1β, IL-17, and TNF-α in the CLP group were significantly higher than those in the Sham group, and the Chiu's score was also significantly higher, indicating that successful modeling altered the permeability of the rat intestinal mucosal barrier, exacerbated mucosal tissue pathological damage, and intensified the inflammatory response. However, after intervention with the composition, the serum levels of D-lactic acid, DAO, endotoxin, IL-1β, IL-17, and TNF-α in the BBR group were lower than those in the CLP group, and the Chiu's score was also lower, verifying that the composition can alleviate intestinal mucosal epithelial cell damage and inhibit the inflammatory response in sepsis-affected rats. Therefore, the composition prepared in this invention can alleviate intestinal mucosal epithelial cell damage in sepsis-affected rats.

[0026] In the field of traditional Chinese medicine, the symptoms of sepsis are described as "stagnation of vital energy and intense heat and toxins".

[0027] Among the components added in this invention, Andrographis paniculata clears heat and detoxifies, cools the blood and reduces swelling, and can improve symptoms such as internal accumulation of pathogenic toxins, fever from colds, sore throat, and mouth ulcers. Forsythia suspensa clears heat and detoxifies, reduces swelling and dissipates nodules, and disperses wind-heat. Anemarrhena asphodeloides clears heat and removes fire, nourishes yin and moistens dryness. It is mainly used to treat exogenous febrile diseases, high fever and thirst, cough due to lung heat, dry cough due to yin deficiency, bone steaming fever, and constipation due to yin deficiency and intestinal dryness. As the principal herb, it exerts the effect of clearing heat and detoxifying.

[0028] Wang Bu Liu Xing (Semen Vaccariae) has the effects of promoting blood circulation, diuresis, and relieving strangury. Fu Niu Hua (Flos Achyranthis Bidentatae) has the effects of dispelling wind and dampness, promoting blood circulation, and relieving pain. Xuan Shen (Radix Scrophulariae) clears heat and cools the blood, nourishes yin and reduces fire, and detoxifies and dissipates nodules. As an assistant herb, it can promote blood circulation and

[0029] Coptis chinensis clears heat and dries dampness, drains fire and detoxifies. Melia toosendan is the dried, mature fruit of Melia toosendan, a plant in the Meliaceae family. It is bitter, cold in nature, and slightly toxic. It enters the liver, small intestine, and bladder meridians and has the effects of soothing the liver, purging heat, regulating qi, and relieving pain. As an adjuvant, it can clear heat from the heart.

[0030] Trillium has the effects of promoting blood circulation, removing blood stasis, soothing the liver and regulating qi, stopping bleeding and relieving pain. Peach leaves clear heat and detoxify, and also contain lycopene. The combination of trillium and peach leaves helps the regeneration and repair of intestinal mucosa after damage, and can also reduce the damage of intestinal mucosal epithelial cells in septic rats. The two are used as adjuvant drugs. All components work together to achieve the effects of clearing heat and detoxifying, promoting blood circulation and relieving heat. Detailed Implementation

[0031] This invention provides a pharmaceutical composition for treating sepsis, prepared from components comprising the following parts by weight: 20-26 parts of Andrographis paniculata, 18-24 parts of Forsythia suspensa, 17-21 parts of Anemarrhena asphodeloides, 15-19 parts of Vaccaria segetalis, 14-18 parts of Achyranthes bidentata, 12-16 parts of Scrophularia ningpoensis, 10-14 parts of Coptis chinensis, 7-11 parts of Melia toosendan, 5-9 parts of Prunus persica leaf, and 3-7 parts of Trillium tectorum; preferably prepared from components comprising the following parts by weight: 21-25 parts of Andrographis paniculata, 19-23 parts of Forsythia suspensa, 18-20 parts of Anemarrhena asphodeloides, 16-18 parts of Vaccaria segetalis, 15-17 parts of Achyranthes bidentata, 13-15 parts of Scrophularia ningpoensis, and 10-14 parts of Coptis chinensis, 7-11 parts of Melia toosendan, 5-9 parts of Prunus persica leaf, and 3-7 parts of Trillium tectorum; The ingredients are: 11-13 parts of *Andrographis paniculata*, 8-10 parts of *Melia toosendan*, 6-8 parts of peach leaf, and 4-6 parts of *Tripterygium wilfordii*. More preferably, the ingredients are prepared from the following components in parts by weight: 22-24 parts of *Andrographis paniculata*, 20-22 parts of *Forsythia suspensa*, 19 parts of *Anemarrhena asphodeloides*, 17 parts of *Vaccaria segetalis*, 16 parts of *Achyranthes bidentata*, 14 parts of *Scrophularia ningpoensis*, 12 parts of *Coptis chinensis*, 9 parts of *Melia toosendan*, 7 parts of peach leaf, and 5 parts of *Tripterygium wilfordii*. Even more preferably, the ingredients are prepared from the following components in parts by weight: 23 parts of *Andrographis paniculata*, 21 parts of *Forsythia suspensa*, 19 parts of *Anemarrhena asphodeloides*, 17 parts of *Vaccaria segetalis*, 16 parts of *Achyranthes bidentata*, 14 parts of *Scrophularia ningpoensis*, 12 parts of *Coptis chinensis*, 9 parts of *Melia toosendan*, 7 parts of peach leaf, and 5 parts of *Tripterygium wilfordii*.

[0032] The present invention also provides a method for preparing the pharmaceutical composition for treating sepsis, comprising the following steps:

[0033] (1) Mix Andrographis paniculata, Forsythia suspensa, Anemarrhena asphodeloides, Vaccaria segetalis, Achyranthes bidentata, Scrophularia ningpoensis, Coptis chinensis, Melia toosendan, Prunus persica leaves and Trillium truncata to obtain material 1;

[0034] (2) Mix the material 1, mixed enzyme and water for enzymatic hydrolysis, inactivate the enzyme, filter, centrifuge and take the supernatant;

[0035] (3) The supernatant is concentrated and dried to obtain a pharmaceutical composition for treating sepsis.

[0036] In this invention, the mass ratio of material 1, mixed enzyme and water in step (2) is 8-12:0.3-0.5:100; preferably 9-11:0.3-0.5:100; further preferably 10:0.3-0.5:100; more preferably 10:0.4:100.

[0037] In this invention, the mixed enzyme is cellulase, protease and α-amylase; the mass ratio of cellulase, protease and α-amylase is 12-16:5-9:7-11; preferably 13-15:6-8:8-10; more preferably 14:7:9.

[0038] In this invention, the enzyme activities of cellulase, protease and α-amylase are all 3200-3800 U / g; preferably 3300-3700 U / g; more preferably 3400-3600 U / g; and even more preferably 3500 U / g.

[0039] In this invention, the enzymatic hydrolysis temperature in step (2) is 42-48°C and the time is 2.5-3.5h; preferably, the enzymatic hydrolysis temperature is 43-47°C and the time is 3h; more preferably, the enzymatic hydrolysis temperature is 44-46°C and the time is 3h; more preferably, the enzymatic hydrolysis temperature is 45°C and the time is 3h.

[0040] In this invention, the centrifugation speed in step (2) is 2200-2800 rpm and the time is 5-9 min; preferably, the centrifugation speed is 2300-2700 rpm and the time is 6-8 min; more preferably, the centrifugation speed is 2400-2600 rpm and the time is 7 min; more preferably, the centrifugation speed is 2500 rpm and the time is 7 min.

[0041] In this invention, the supernatant in step (3) is concentrated to 1 / 5 to 1 / 3 of its original volume; preferably 1 / 4.

[0042] In this invention, the drying in step (3) is spray drying.

[0043] In this invention, the inlet air temperature of the spray dryer is 128-136°C and the outlet air temperature is 67-73°C; preferably, the inlet air temperature of the spray dryer is 129-135°C and the outlet air temperature is 68-72°C; more preferably, the inlet air temperature of the spray dryer is 130-134°C and the outlet air temperature is 69-71°C; even more preferably, the inlet air temperature of the spray dryer is 132°C and the outlet air temperature is 70°C.

[0044] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0045] Example 1

[0046] A pharmaceutical composition for treating sepsis is prepared from the following components in parts by weight: 20 parts of Andrographis paniculata, 18 parts of Forsythia suspensa, 17 parts of Anemarrhena asphodeloides, 15 parts of Vaccaria segetalis, 14 parts of Achyranthes bidentata, 12 parts of Scrophularia ningpoensis, 10 parts of Coptis chinensis, 7 parts of Melia toosendan, 5 parts of Prunus persica and 3 parts of Trillium truncatum.

[0047] The preparation method of the pharmaceutical composition for treating sepsis includes the following steps:

[0048] (1) Mix Andrographis paniculata, Forsythia suspensa, Anemarrhena asphodeloides, Vaccaria segetalis, Achyranthes bidentata, Scrophularia ningpoensis, Coptis chinensis, Melia toosendan, Prunus persica leaves and Trillium truncata to obtain material 1;

[0049] (2) Mix the material 1, mixed enzyme and water at a mass ratio of 8:0.3:100, enzymatically hydrolyze at 42℃ for 2.5h, filter after enzyme inactivation, centrifuge at 2200rpm for 5min, and take the supernatant;

[0050] The mixed enzyme consists of cellulase, protease and α-amylase in a mass ratio of 12:5:7, and all enzyme activities are 3200 U / g.

[0051] (3) The supernatant is concentrated to 1 / 5 of its original volume and spray-dried (inlet air temperature 128°C, outlet air temperature 67°C) to obtain a pharmaceutical composition for treating sepsis.

[0052] Example 2

[0053] A pharmaceutical composition for treating sepsis is prepared from the following components in parts by weight: 26 parts of Andrographis paniculata, 24 parts of Forsythia suspensa, 21 parts of Anemarrhena asphodeloides, 19 parts of Vaccaria segetalis, 18 parts of Achyranthes bidentata, 16 parts of Scrophularia ningpoensis, 14 parts of Coptis chinensis, 11 parts of Melia toosendan, 9 parts of Prunus persica and 7 parts of Tripterygium wilfordii.

[0054] The preparation method of the pharmaceutical composition for treating sepsis includes the following steps:

[0055] (1) Mix Andrographis paniculata, Forsythia suspensa, Anemarrhena asphodeloides, Vaccaria segetalis, Achyranthes bidentata, Scrophularia ningpoensis, Coptis chinensis, Melia toosendan, Prunus persica leaves and Trillium truncata to obtain material 1;

[0056] (2) Mix the material 1, mixed enzyme and water at a mass ratio of 12:0.5:100, enzymatically hydrolyze at 48℃ for 3.5h, filter after enzyme inactivation, centrifuge at 2800rpm for 9min, and take the supernatant;

[0057] The mixed enzyme consists of cellulase, protease and α-amylase in a mass ratio of 16:9:11, and all enzyme activities are 3800 U / g.

[0058] (3) The supernatant is concentrated to 1 / 3 of its original volume and spray-dried (inlet air temperature 136°C, outlet air temperature 73°C) to obtain a pharmaceutical composition for treating sepsis.

[0059] Example 3

[0060] A pharmaceutical composition for treating sepsis is prepared from the following components in parts by weight: 23 parts of Andrographis paniculata, 21 parts of Forsythia suspensa, 19 parts of Anemarrhena asphodeloides, 17 parts of Vaccaria segetalis, 16 parts of Achyranthes bidentata, 14 parts of Scrophularia ningpoensis, 12 parts of Coptis chinensis, 9 parts of Melia toosendan, 7 parts of Prunus persica leaf, and 5 parts of Trillium truncatum.

[0061] The preparation method of the pharmaceutical composition for treating sepsis includes the following steps:

[0062] (1) Mix Andrographis paniculata, Forsythia suspensa, Anemarrhena asphodeloides, Vaccaria segetalis, Achyranthes bidentata, Scrophularia ningpoensis, Coptis chinensis, Melia toosendan, Prunus persica leaves and Trillium truncata to obtain material 1;

[0063] (2) Mix the material 1, mixed enzyme and water at a mass ratio of 10:0.4:100, enzymatically hydrolyze at 45℃ for 3h, filter after enzyme inactivation, centrifuge at 2500rpm for 7min, and take the supernatant.

[0064] The mixed enzyme consists of cellulase, protease and α-amylase in a mass ratio of 14:7:9, and all enzyme activities are 3500 U / g.

[0065] (3) The supernatant is concentrated to 1 / 4 of its original volume and spray-dried (inlet air temperature 132°C, outlet air temperature 70°C) to obtain a pharmaceutical composition for treating sepsis.

[0066] Experimental Example 1

[0067] 1. Materials and Methods

[0068] 1.1 Experimental animals: 6 - 8 - week - old SPF - grade male SD rats weighing 200 ± 20 g were purchased from Shandong Pengyue Experimental Animal Company. The production license number of experimental animals is: SCXK(Shandong)20190003. The experimental animals were housed in the Animal Experiment Center of the Affiliated Hospital of Shandong University of Traditional Chinese Medicine. They were adaptively fed for one week before the experiment, with free access to water and food. The feeding and experimental protocols of the experimental animals were approved by the Animal Experiment Ethics Committee of the Affiliated Hospital of Shandong University of Traditional Chinese Medicine, approval number: AWE - 2019 - 026.

[0069] 1.2 Test drugs and instruments: Dimethyl sulfoxide (DMSO, D8370), Beijing Solarbio Science & Technology Co., Ltd.; DAO kit (E - EL - R0331c), Rat TNF - α ELISA Kit (E - EL - R2856c), Rat IL - 1β ELISA Kit (E - EL - R0012c), Rat IL - 17A ELISA Kit (E - EL - R0566c), Elabscience; D - lactate kit (SBJ - 80192), endotoxin kit (SBJ - 80392), Nanjing Sbjbio Technology Co., Ltd.; microplate reader Muliskan, low - speed automatic balance centrifuge ST8 type, Thermo Scientific, USA; rotary microtome RM2235 type, LEICA, Germany; pathological tissue baking instrument PHY - III type, Changzhou Zhongwei Electronic Instrument Co., Ltd.; microscope DX45 type, OLYMPUS, Japan.

[0070] 1.3 Grouping and model establishment: Thirty SD rats were randomly divided into 3 groups. Sham operation group (Sham group), CLP model group (CLP group), experimental group (BBR group), with 10 rats in each group. Rats in the Sham group only underwent laparotomy and received DMSO / aqueous solution pretreatment; CLP model group: underwent CLP model establishment and received DMSO / aqueous solution pretreatment; experimental group: underwent CLP model establishment and received composition pretreatment.

[0071] The cecal ligation and puncture (CLP) method was used to prepare a sepsis rat model: Rats were fasted routinely for 8 h before surgery. Before model establishment, intraperitoneal anesthesia (10% chloral hydrate 15 ml / kg) was first performed. After sufficient anesthesia, the skin of the middle and lower abdomen was routinely prepared and disinfected. A 1 - cm incision was first made along the mid - abdominal line, and then the abdomen was opened layer by layer to enter the abdominal cavity. The cecum was pulled out of the incision to the outside of the abdominal cavity, and the proximal 1 / 3 of the cecum near the ileocecal valve was ligated. At the mid - point between the ligation site and the cecal blind end, a 20G sterile needle was used to penetrate and puncture, and a little intestinal content was extruded. Then, the cecum was returned to the abdominal cavity and the abdomen was closed. After the operation, all rats were injected subcutaneously in the neck with 37°C normal saline at a dose of 2 ml / 100 g for fluid resuscitation, and they were allowed free access to food and water after the operation.

[0072] 1.4 Administration Method: The product obtained in Example 3 was dissolved in DMSO at a ratio of 1g:2ml, and then the solution was diluted with distilled water at a ratio of 3:7. Each group was administered the corresponding diluted solution by gavage at a rate of 100mg / kg at 72h, 48h, 24h, and 0h before modeling. The sham-operated group and the CLP model group were administered only DMSO / water solution by gavage, while the experimental groups were administered DMSO / water solution containing the product of Example 3 by gavage.

[0073] 1.5 Specimen Collection and Detection Twenty-four hours after modeling, rats were anesthetized with 10% chloral hydrate. Following anesthesia, the abdomen was opened, and blood was first drawn from the abdominal aorta. The collected blood was centrifuged at 2500 rpm for 20 minutes, and the serum was separated and collected, then stored at -80°C for later testing. Tissue from the terminal ileum of the rats was taken, placed in cryovials, and stored at -80°C for later testing. Simultaneously, a portion of the ileum tissue was fixed with 4% paraformaldehyde for pathological examination.

[0074] After processing rat serum samples according to relevant procedures, D-lactic acid, diamine oxidase (DAO), and endotoxin levels were measured in rat serum following the instructions of the kits for D-lactic acid, DAO, and endotoxin, using steps such as sample loading, incubation, solution preparation, washing, enzyme addition, color development, termination, and measurement. This was used to determine whether changes in intestinal barrier permeability occurred in septic rats. Simultaneously, rat serum was collected and IL-1β, IL-17, and TNFα were measured in rat serum according to the instructions of the ELISA kit, following the steps of sample loading, incubation, washing, enzyme addition, color development, termination, and measurement, to clarify the effect of the combined assay on the expression of inflammatory factors in the serum of septic rats. Rat ileal tissue fixed in 4% paraformaldehyde was dehydrated, cleared, paraffin-embedded, embedded, sectioned, baked, and stained with hematoxylin and eosin (HE). The pathological damage of the ileal tissue was observed under a light microscope, and Chiu's intestinal mucosal damage score was performed.

[0075] 1.6 Statistical Analysis SPSS 20.0 software was used for statistical analysis. Quantitative data were expressed as mean ± standard deviation (X±S). One-way ANOVA was used for comparisons between groups. P < 0.05 was considered statistically significant.

[0076] 2. Results

[0077] 2.1 Effects of the composition on serum D-lactic acid, DAO, and endotoxin in septic rats

[0078] After cecal ligation modeling, the serum D-lactic acid, DAO, and endotoxin levels in the CLP group rats were significantly higher than those in the Sham group (P < 0.05); while the serum D-lactic acid, DAO, and endotoxin levels in the BBR group rats were significantly lower than those in the CLP group (P < 0.05).

[0079] Table 1 Comparison of serum D-lactic acid, DAO, and endotoxin levels in rats of different groups (pg / ml) )

[0080]

[0081] Note: Compared with the Sham group * P < 0.05; compared with the CLP group, △ P < 0.05. The same applies below.

[0082] 2.2 Effects of the composition on intestinal histopathology in septic rats

[0083] Using light microscopy, the pathological damage of the terminal ileum tissue in rats was observed. In the Sham group: the intestinal epithelial structure of the mucosa was intact, the epithelial cells were normal in morphology and tightly arranged, the lamina propria had abundant intestinal glands with numerous goblet cells, and no obvious inflammation was observed. In the CLP group: large areas of epithelial cell shedding were observed in the mucosa, exposing the lamina propria; local ulcers were visible, the number of intestinal glands in the lamina propria was reduced and replaced by proliferating connective tissue, accompanied by numerous lymphocyte infiltration; local venous congestion and dilation were observed; the Chiu's score was significantly higher than that in the Sham group (P < 0.05). In the BBR group: a small number of epithelial cells were observed locally in the mucosa; the lamina propria had abundant intestinal glands with numerous goblet cells, and a small amount of inflammatory infiltration was observed locally; the Chiu's score was significantly lower than that in the CLP group (P < 0.05).

[0084] Table 2 Comparison of Chiu's scores for pathological damage to rat intestinal mucosa in different groups (points, )

[0085]

[0086] 2.3 Effects of the composition on serum IL-1β, IL-17 and TNF-α in septic rats

[0087] The serum levels of IL-1β, IL-17, and TNF-α in the CLP group rats were significantly higher than those in the Sham group (P < 0.05); while the serum levels of IL-1β, IL-17, and TNF-α in the BBR group rats were significantly lower than those in the CLP group (P < 0.05).

[0088] Table 3 Comparison of serum IL-1β, IL-17 and TNF-α levels in rats of different groups (pg / ml) )

[0089]

[0090]

[0091] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A pharmaceutical composition for treating sepsis, characterized in that, It is prepared from the following components in parts by weight: Andrographis paniculata 20-26 parts, Forsythia suspensa 18-24 parts, Anemarrhena asphodeloides 17-21 parts, Vaccaria segetalis 15-19 parts, Achyranthes bidentata 14-18 parts, Scrophularia ningpoensis 12-16 parts, Coptis chinensis 10-14 parts, Melia toosendan 7-11 parts, Prunus persica leaf 5-9 parts, and Trillium truncata 3-7 parts.

2. The method for preparing the pharmaceutical composition for treating sepsis according to claim 1, characterized in that, Includes the following steps: (1) Mix Andrographis paniculata, Forsythia suspensa, Anemarrhena asphodeloides, Vaccaria segetalis, Achyranthes bidentata, Scrophularia ningpoensis, Coptis chinensis, Melia toosendan, Prunus persica leaves and Trillium truncata to obtain material 1; (2) Mix the material 1, mixed enzyme and water for enzymatic hydrolysis, inactivate the enzyme, filter, centrifuge, and take the supernatant; (3) The supernatant is concentrated and dried to obtain a pharmaceutical composition for treating sepsis.

3. The preparation method according to claim 2, characterized in that, In step (2), the mass ratio of material 1, mixed enzyme and water is 8~12:0.3~0.5:

100.

4. The preparation method according to claim 3, characterized in that, The mixed enzyme is cellulase, protease and α-amylase; the mass ratio of cellulase, protease and α-amylase is 12~16:5~9:7~11.

5. The preparation method according to claim 4, characterized in that, The enzyme activities of the cellulase, protease, and α-amylase are all 3200~3800 U / g.

6. The preparation method according to claim 2, characterized in that, The enzymatic hydrolysis in step (2) is carried out at a temperature of 42~48℃ for 2.5~3.5h.

7. The preparation method according to claim 2, characterized in that, In step (2), the centrifugation speed is 2200~2800 rpm and the time is 5~9 min.

8. The preparation method according to claim 2, characterized in that, In step (3), the supernatant is concentrated to 1 / 5 to 1 / 3 of its original volume.

9. The preparation method according to claim 2, characterized in that, The drying in step (3) is spray drying; the inlet air temperature of the spray drying is 128~136℃ and the outlet air temperature is 67~73℃.

10. The use of a pharmaceutical composition according to claim 1 or a pharmaceutical composition prepared by any one of claims 2 to 9 in the preparation of a drug for treating sepsis.

Images