WO2008154795A1

WO2008154795A1 - Use of sanchiosides for treating septicemia

Info

Publication number: WO2008154795A1
Application number: PCT/CN2008/000877
Authority: WO
Inventors: Jingyan Han; Kai Sun; Jun Guo; Chuanshe Wang; Yuying Liu
Original assignee: Tianjin Tasly Pharmaceutical Co. Ltd
Priority date: 2007-06-21
Filing date: 2008-04-29
Publication date: 2008-12-24
Also published as: CN101327236A; CN101327236B

Abstract

The invention discloses the use of sanchinosides, particularly ginsenoside Rb1, ginsenoside Rg1, and notoginsenoside R1 for treating and/or preventing septicemia.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a novel use of a traditional Chinese medicine product, and particularly to the use of the notoginsenoside components Rbl, Rgl and R1 for the treatment and/or prevention of sepsis. Background technique

Panax notoginseng (PN) is the root of the traditional Chinese medicine Burk F. H. Chen after air drying. Panax notoginseng saponins (also known as: Panax Notoginsenos ides, PNS) are the main active ingredients of Panax notoginseng. They are composed of more than 30 different saponins and are widely used clinically to treat microcirculatory disorders. Diseases such as cardiovascular disease and liver dysfunction.

Panax notoginseng saponins have been listed in the National Pharmacopoeia, and there are corresponding preparation products listed, such as: Xuesaitong for injection. Ginsenoside Rbl, referred to as Rbl [English name] Ginsenoside Rbl [chemical name] β -D-Glucopyranoside,

(3 β, 12 ) -20- [ (6-0- β -D-glucopyranosyl- β -D-glucopyranosyl) oxy] - 12_h ydroxydammar-24-3n-3yl 2-0 β -D-glucopyranosyl- [ Molecular Formula] C54H92023 [Molecular weight] 1109. 26

Ginsenoside Rgl, referred to as Rgl, [English name] Ginsenoside Rgl [alias] panaxoside A, Sanchinoside CI [chemical name] β - D-Glucopyranoside, (3 β , 6 α , 12 β ) -3, 12-dihydroxydammar-24- Ene-6, 20-diylbis- [Molecular formula] C42H72014o2H20. Notoginsenoside R1 [referred to as R" [English name] Notoginsenoside R1

These three saponins are the main active ingredients of Panax notoginseng which has been widely used in traditional Chinese medicine and have a blood circulation.

Sepsis is an acute systemic infection caused by pathogenic bacteria or conditional pathogens invading the blood circulation, producing toxins and other metabolites. It is characterized by chills, fever, rash, joint pain and hepatosplenomegaly. There may be septic shock and migratory lesions. The causes of sepsis are:

First, the human factor:

1 When the skin and mucous membranes are damaged or have suppurative inflammation, the bacteria easily invade the body. 2 The human immune response can be divided into two types: non-specific immune response and specific immune response, and the latter can be divided into cellular immunity and humoral immunity. When the immune function of the body declines, the effect of phagocytosis and killing of bacteria cannot be fully exerted. Even if the amount of invading bacteria is small, the pathogenicity is not strong and can cause sepsis.

3 The iatrogenic infection caused by the conditional pathogens also gradually increased.

Second, the bacterial factors:

Mainly related to the virulence and quantity of pathogens. Pathogenic bacteria with high virulence or a large number of bacteria enter the body, which is more likely to cause sepsis.

The main symptoms of sepsis are -

1. Primary inflammation: The primary inflammation caused by various pathogens is related to its distribution in the human body. Primary inflammation is characterized by localized redness, swelling, heat, pain, and dysfunction.

2. Symptoms of Toxemia: The onset is more rapid. Often there are chills, high fever, and fever, which are mostly relaxation heat or intermittent heat. They can also be caused by heat retention, irregular heat and bimodal fever. The latter are caused by sepsis-negative bacilli sepsis. The fever is accompanied by varying degrees of symptoms of toxemia such as headache, nausea, vomiting, bloating, abdominal pain, generalizedness, and muscle and joint pain.

3. Rash: Seen in some patients, the most common defects are distributed in the trunk, limbs, conjunctiva, oral mucosa, etc., not many.

4. Joint symptoms: There may be large joint redness, swelling, heat, pain and limited mobility, even complicated with joint fluid and empyema. It is more common in the course of sepsis such as Gram-positive cocci, meningococcal, and Alcaligenes. .

5. Infectious shock: About 1/5~1/3 of patients with sepsis, manifested as irritability, rapid pulse rate, cold limbs, skin spots, decreased urine output and decreased blood pressure, etc. Caused by severe toxemia.

6. Hepatosplenomegaly: Generally only mild swelling.

According to the study, lipopolysaccharide (LPS) is one of the components of the cell wall of Gram-positive bacteria, causing multiple manifestations of human sepsis and septic shock caused by Gram-positive bacteria. The microcirculatory disorder that occurs during sepsis is a systemic inflammatory response caused by excessive release of LPS, mediated by a number of activated factors such as adhesion molecules, reactive oxygen species (R0S), and inflammatory precursor mediators such as tumors. Necrosis factor-a (TNF_a), interleukin- ₆ (IL-6), and interferon-Y (INF-γ). The microcirculatory disorder induced by LPS plays a key role in organ dysfunction during sepsis. During this process, white blood cells rotate along the endothelium and adhere to the vascular endothelium. Hydrogen peroxide and mast cell degranulation are produced in the vessel wall. Newspaper The key steps of the road.

The main clinical treatments for current sepsis include volume recovery, the use of catecholamines, and antibiotic-compensatory treatment. These methods can increase the survival rate of patients with sepsis, but clinically lead to an increase in blood sugar and a decrease in blood pressure and blood calcium, and bleeding. Clinical studies suggest that anti-TNF-α therapy may be helpful in the treatment of sepsis. However, recent studies have shown that this method does not cure sepsis and increases mortality in patients with severe sepsis. Therefore, finding a way to prevent severe microcirculation in LPS-induced sepsis remains a challenge.

The present inventors have found that three saponins Rbl, Rgl and R1 in Panax notoginseng can reduce the decrease of the movement speed of red blood cells caused by injection of LPS, and Rbl, Rgl and R1 can reduce the number of adherent cells. , inhibit mast cell degranulation and inhibit the increase of cytokine levels. In vitro experiments using flow cytometry further confirmed that Rbl and Rgl can significantly inhibit the increase of CD1 lb/CD18 expression in neutrophils induced by LPS, while Rbl and Rgl can inhibit LPS stimulation and cause H ₂ 0 ₂ from neutrophils. Release. The above results indicate that the three saponins Rbl, Rgl and R1 can improve LPS-induced microcirculatory disorders. It has obvious therapeutic effects on sepsis. Summary of the invention:

The present invention provides a novel therapeutic use of a notoginsenoside component. The new therapeutic use is the treatment of sepsis caused by microcirculatory disorders with a component of saponins.

To this end, the present invention provides a novel use of a medicament for preparing a medicament for treating and/or preventing sepsis using a component of saponins.

The panax notoginseng saponin component of the present invention is selected from the group consisting of ginsenoside Rbl, ginsenoside Rgl and notoginsenoside

Rl.

The three saponin components of the present invention, ginsenoside Rbl, ginsenoside Rgl and notoginsenoside R1, are available in the prior art and are commercially available or can be prepared according to the prior art and meet pharmaceutical standards. Preferably, the purity is > 50%, more preferably > 90%, most preferably > 98%.

The medicament prepared by the present invention is a pharmaceutical preparation composition prepared by using the above-mentioned notoginsenoside component as a pharmaceutically active ingredient.

The ninth, the remainder is 0. 1-99. 9%, the remainder is 0. 1-99. 9%, the remainder is A pharmaceutically acceptable carrier. The pharmaceutical preparation composition of the present invention is in the form of a unit dosage form, which means a unit of the preparation, such as each tablet of the tablet, each capsule of the capsule, each bottle of the oral solution, granules per bag, injection Every one of them.

The pharmaceutical preparation composition of the present invention may be in any pharmaceutically acceptable dosage form, and includes: a tablet, a sugar-coated tablet, a film-coated tablet, an enteric coated tablet, a capsule, a hard capsule, a soft capsule, orally. Liquid, buccal, granule, granule, pill, powder, ointment, dandruff, suspension, powder, solution, injection, suppository, ointment, plaster, cream, spray, drops, patch .

The traditional Chinese medicine preparation of the present invention, the preparation for oral administration may contain common excipients such as a binder, a filler, a diluent, a tablet, a lubricant, a disintegrant, a coloring agent, a flavoring agent and a wetting agent. , The tablets can be coated if necessary.

Suitable fillers include cellulose, mannitol, lactose and other similar fillers. Suitable disintegrating agents include starch, polyvinylpyrrolidone and starch derivatives such as sodium starch glycolate. Suitable lubricants include, for example, magnesium stearate. Suitable pharmaceutically acceptable wetting agents include sodium decyl sulfate.

The solid oral composition can be prepared by a usual method such as mixing, filling, tableting or the like. Repeated mixing allows the active material to be distributed throughout those compositions that use large amounts of filler.

The oral liquid preparation may be in the form of, for example, an aqueous or oily suspension, solution, emulsion, syrup or elixir, or may be a dry product which may be formulated with water or other suitable carrier before use. Such liquid preparations may contain conventional additives such as suspending agents such as sorbitol, syrup, methylcellulose, gelatin, hydroxyethylcellulose, carboxymethylcellulose, aluminum stearate or hydrogenated edible fats. Emulsifiers such as lecithin, sorbitan monooleate or gum arabic; non-aqueous carriers (which may include edible oils), such as almond oil, fractionated coconut oil, oily esters of esters such as glycerol, propylene glycol or ethanol; The agent, for example, p-hydroxybenzyl or propylparaben or sorbic acid, and if desired, may contain conventional flavoring or coloring agents.

For injection, the liquid unit dosage form prepared contains the active substance of the invention and a sterile vehicle. This compound can be suspended or dissolved depending on the carrier and concentration. The solution is usually prepared by dissolving the active substance in a carrier, sterilizing it by filtration before filling it into a suitable vial or ampoule, and then sealing. Excipients such as a local anesthetic, preservative and buffer may also be dissolved in such a carrier. To increase its stability, the composition can be frozen after filling the vial and the water removed under vacuum.

The traditional Chinese medicine preparation of the invention can be selectively added to a suitable pharmaceutically acceptable load when preparing the medicament The pharmaceutically acceptable carrier is selected from the group consisting of: mannitol, sorbitol, sodium metabisulfite, sodium hydrogen sulfite, sodium thiosulfate, cysteine hydrochloride, thioglycolic acid, methionine, vitamins (:, disodium EDTA, EDTA Calcium sodium, monovalent alkali metal carbonate, acetate, phosphate or its aqueous solution, hydrochloric acid, acetic acid, sulfuric acid, phosphoric acid, amino acid, sodium chloride, potassium chloride, sodium lactate, xylitol, maltose, glucose , fructose, dextran, glycine, starch, sucrose, lactose, mannitol, silicon derivatives, cellulose and its derivatives, alginate, gelatin, polyvinylpyrrolidone, glycerol, soil temperature 80, agar, Calcium carbonate, calcium hydrogencarbonate, surfactant, polyethylene glycol, cyclodextrin, e-cyclodextrin, phospholipid material, kaolin, talc, calcium stearate, magnesium stearate, and the like.

The preparation of the present invention is used according to the condition of the patient at the time of use, and can be taken three times a day for 1-20 doses, such as: 1-20 bags or tablets or tablets, each dose of lmg-1000 mg. The therapeutic use of the present invention is demonstrated by the following experiments:

Materials and Method

Reagents and test animals

Rbl and Rgl were purchased from the China National Institute for the Control of Pharmaceutical and Biological Products, and R1 was purchased from Kunming Feng-Shan-Jian Pharmaceutical Company. All saponins were purified by affinity chromatography.

Male sprague-dawley (SD) rats weighing between 200 and 250 grams were obtained from the Animal Center of Peking University Medical School. The license number for this batch of animals is SCXK 2002-0001. Before the test, the standard test room mouse feed (Peking University Medical Center Animal Center) was used for 72 hours, the body temperature was maintained at 24 ± 1 °C, the relative humidity was 50 ± 1%, and all the test animals received the 12/12-light/dark cycle. Animals were fasted for 12 hours before the test. Animal treatment was carried out in accordance with the guidelines of the Animal Research Committee of Peking University.

Mesenteric microcirculation

The rats were anesthetized with urethane (1.25 mg/kg body weight), and the rats were intraperitoneally inoculated with a median incision (20-30 mm long for injection of the femoral vein and internal jugular vein.

The ileocecal portion of the mesentery (10-15 cm in the tail of the mesentery) was carefully separated, and the mesentery was removed from the abdominal cavity and laid flat on a transparent plastic table. The temperature and humidity of the mesentery were maintained by continuous perfusion of 37 Ό normal saline on the surface. The hemodynamics of the mesenteric microcirculation were observed using an inverted microscope (Leica, DM-IRB, Germany) using a transilluminator. Install a TV camera (Japan, Toshiba Jk-TU53H) on the microscope and transfer the image to a color display (China, TCL, J2118A), through the DVD player (middle Country, Malata DVR-R25) records all images. In this study, a single branch with no branching and no obvious curvature was selected as the experimental observation. The diameter of the small vein was between 30μηι and 50 μηι, and the length was about 200 μπι.

Administration

The test rats were randomly divided into five groups of six each. For the rats in the LPS group and the control group, the test animals were pre-observed for 10 minutes, followed by continuous administration through the right jugular vein, saline (1 ml/hr, control group), and saponin (Rbl, dissolved in saline). Rgl or R1) (5 mg/kg hr, simple saponin group), and LPS dissolved in saline (2 mg/kg/hr, LPS group) for 60 minutes. For each of the three experimental groups, each group was injected with the saponin Rbl (Rbl + LPS group), Rgl (Rgl + LPS group) or Rl (R1 + LPS group) through the right jugular vein in the same manner as the LPS group. Saponin injection was started 20 minutes before the injection of LPS, and saponin continued to be injected until the end of the observation. The saponins used in each of the three groups were pre-dissolved in saline and then administered at a dose of 5 mg/kg/hr. The hemodynamic status of the mesenteric microcirculation of each group was continuously observed and recorded.

Determination of vessel diameter

Blood vessel diameters were determined using Image-Pro Plus 5.0 (Media Cybernetic, USA) software for LPS injections (0 minutes) and images recorded at 20 minutes, 40 minutes, and 60 minutes after LPS injection. The diameter of the blood vessel was measured three times at the same position and then averaged.

Determination of red blood cell rate

Convert the monitor's CCD to a high-speed TV camera system (FASTCAM-ultima APX, photron, Japan) to record the rate of red blood cells in the venule at 2000 frames/s, and then store the stored high-speed images at a rate of 25 frames per s. Play. Image-Pro Plus 5.0 software (Media Cybernetic, USA) was used to measure the rate of red blood cells in the venules at the time of LPS injection (0 minutes) and at 20 minutes, 40 minutes, and 60 minutes after LPS injection.

Shear rate determination

The shear rate of the venous vessel wall was estimated using the formula SR=2.12x8x[V/D], where D represents the mean diameter of the vessel, V is the average rate of red blood cells, and 2.12 is the empirical correction factor for the measured microvessel velocity profile in vivo. .

Determination of leukocyte adhesion

Reviewing the dynamic behavior of white blood cells by replaying the recorded images to evaluate white blood cell pairs Adhesion of the vessel wall. In this study, adherent white blood cells were defined as adhering to the ipsilateral vascular wall for longer than

10s of cells. At the time of LPS injection (0 minutes) and 20 minutes, 40 minutes, and 60 minutes after LPS injection, a certain number of blood vessel fragments (30~50 μπι in diameter and 200 μιη in diameter) were counted by randomly selecting the recorded images. The number of cells.

Determination of leukocyte migration

The number of leukocytes migrated was estimated by reviewing the white blood cell images recorded at the LPS injection (0 minutes) and at 20 minutes, 40 minutes, and 60 minutes after the LPS injection. The migrated white blood cells are defined as the number of white blood cells around a certain length of venules. The number of white blood cells in the surrounding tissues (30 to 50 μm in diameter) is determined by randomly selecting the recorded images and selecting a blood vessel of 200 μηι in length (30 to 50 in diameter). Ππι). Determination of mast cell degranulation

60 minutes after the injection of LPS, the tissue was perfused with 0.1% toluidine blue for 1 minute and then bleached with saline. Cells in which intracellular particles are released into the surrounding tissue are considered to be degranulated mast cells, and the number of cells in the circular field of view of the microscope microscope is counted to determine the number of such cells. Each of the mesenteric windows was evaluated by selecting five fields of view around the microvasculature. The number of degranulated and non-degranulated mast cells was recorded, and then the proportion of degranulated mast cells was calculated.

Determination of cytokines

After observing the mesenteric microcirculation, blood samples from each test animal were collected via the abdominal aorta route for heparin anticoagulation (20 unit/ml blood). The plasma is then separated by centrifugation. 50 μL of plasma and standard substances were incubated with 50 μΐ of the attracted beads for 1 hour at room temperature, then mixed with 50 μΐ of TNF-α and IL-6 detection antibodies, and incubated for 2 hours at room temperature to form a sandwich complex. . After co-incubation, 1 ml of wash buffer (BD Biosciences Pharmingen, USA) was added to each tube and the mean fluorescence intensity was measured using a flow cytometer (FACS Calibur, B.D. Co, USA). Data analysis was performed using BD Cytometric Bead Array analysis software.

In vitro assay of CD11b/CD18 expression

An in vitro assay of CDl lb/CD18 expression was performed on another group (n=6) of SD rats. Rats were fasted for 12 hours before the test. Rats were anesthetized with urethane (1.25 mg/kg body weight, intramuscularly). Blood was taken from the abdominal aorta of each test animal, then anticoagulated with heparin (20 units/ml of blood), and then 20 samples were prepared as follows: Control, Rbl (0.2 mg/ml), Rgl (0.2 mg) /ml), Rl (0.2 mg/ml), Rbl (1.0 mg/ml), Rgl (1.0 mg/ml), Rl (1.0 mg/ml), Rbl (2.0 mg/ml), Rgl (2.0 mg/ml) ), Rl (2.0 mg/ml), LPS, Rbl+LPS (0.2 mg/ml), Rgl+LPS (0.2 mg/ml): Rl+LPS (0.2 mg/ml), Rbl+LPS (1.0 mg/ml), Rgl+LPS (1.0 mg/ml), Rl+LPS (1.0 mg/ml), Rbl+LPS (2.0 mg/ml), Rgl+LPS (2.0 mg/ml) and Rl+LPS (2.0 mg/ml). 0.2 ml of blood was added to each of the prepared samples. The control group was 8 μΐ saline. The single saponin group consisted of one of 4 μΐ saline and 4 μΐ of three saponins, and the final concentration was found in the above parentheses. The LPS sample consisted of 4 μΐ saline and 4 μΐ LPS, and finally the concentration of LPS reached 2 _μβ /ηι1 _0. All other samples consisted of one of 4 μΐ LPS and 4 μΐ of three saponins, finally achieving the concentration of LPS. 2 μ£/ιη1, the final concentration of each test saponin is shown in parentheses above (Sun et al., 2006). Mix the sample at 37 °. It was stored for 2 hours and then co-incubated with lugFITC-labeled anti-CDl lb antibody or anti-CD18 antibody (BD Biosciences Pharmingen, USA) for 20 minutes at room temperature. The cells were solubilized with hemolysin (BD Biosciences Immunocytometer Systems, USA) and then washed twice with PBS. The mean fluorescence intensity was determined using a flow cytometer (FACS Calibur, BDCo, USA). Neutrophils were selected using the FSC-SSC scatter plot. The number of neutrophils of 5,000 was determined for each sample, and then the average fluorescence intensity was calculated.

Analysis of H ₂ 0 ₂ and '0 ₂ production in neutrophil cells in vitro

Blood was taken from the abdominal aorta and then heparin was used for anticoagulation. Neutrophils were isolated using a mono-poly dissolution medium as described by Ting (Ting and Morris, 1971). The cell suspension was then prepared using PMI 1640 medium according to the modified Shen method (Shen et al., 1998) and then intracellular hydrogen peroxide (H ₂ 0 ₂ ) and superoxide were measured by flow cytometry (BD company, USA). Yield of anion ( ' 0 ₂ — ). The brief procedure is that neutrophils are co-incubated with 0.2 mM 2',7'-dichlorofluorescein diacetate (DCFH-DA) for 5 minutes at 37 °C, again with 0.1 mM ethidium dihydrobromide. (HE) Co-incubation for 15 minutes. The cells were then treated with Rbl (1.0 mg/ml), Rgl (1.0 mg/ml) or Rl (1.0 mg/ml) alone or with LPS (2 g/ml). After co-incubation with ice for 20 minutes, flow cytometry was used to measure the fluorescence of 525 nm emitted by 2',7'-dichlorofluorescein (DCF) and the fluorescence of 590 nm emitted by ethidium bromide (EB). The contents of H ₂ 0 ₂ and '0 ₂ — were determined. Determination of plasma alkaline phosphatase

After observing the microcirculation of the mesentery, blood was taken from the abdominal aorta of each test animal and then anticoagulated with heparin (20 units per ml of blood). Plasma was separated by centrifugation. Plasma alkaline phosphatase concentration was measured using an alkaline biophosphatase kit (Olympus, US) using an automated biochemical analyzer (7170A, Hitachi, Japan).

Statistics Statistical analysis was performed using analysis of variance and F-test. Values are expressed as mean soil SE (N=6). A P value of less than 0.05 is considered statistically significant.

Result

There was no significant difference between the diameters of the mesenteric venules detected at 0 o'clock and other time points after LPS injection. The erythrocyte rate of the mesenteric venules varied with time under the experimental conditions, and the red blood cell rate in the venule of the control group did not change during the observation period of 60 minutes. In contrast, the red blood cell rate of the mesenteric venules of the LPS group was progressively decreased 40 minutes after LPS injection, and was significantly lower than the 0 point and the control group (p < 0.05). At 60 minutes after the injection, the measured rate of red blood cells was 1.30 ± 0.11 mm/sec, which was 70% of the time value of 0 o'clock. Rbl, Rgl or R1 treatment can change the LPS-induced decrease in mesenteric venule erythrocyte rate in a rat to a certain extent.

In the rat mesenteric venule erythrocyte adhesion over time, at 0 o'clock, the number of white blood cells adhering to the venule wall did not differ significantly between these groups. At the end of the observation period of the control group, adherent white blood cells The number has only slightly increased to 1.72 ± 0.71 cells /200 μΓη. In contrast, the LPS group showed a significant linear increase in time and number, from 5.00 ± 0.83 cells / 200 μπι at 20 minutes after LPS injection to a high value of 60 minutes at 12.72 ± 1.41 cells / 200 Ππι, the value of the increase is more than 15 times compared to the 0 o'clock. Administration of Rbl, Rgl or R1 resulted in improved LPS-induced adhesion of leukocytes to the venular wall, although for each saponin there was some variation over time, and inhibition of R1 occurred after LPS injection. At 20 minutes, inhibition of Rbl occurred 40 minutes after LPS injection, while inhibition of Rgl occurred 60 minutes after LPS injection.

The condition in which leukocytes migrate out of the mesenteric venule wall over time is that no obvious white blood cells that migrate out of the vessel wall are detected at any time point. For the control group, no leukocytes that migrated out of the vessel wall were observed during the entire observation period. In contrast, the number of leukocytes that migrated out of the vein wall after injection of LPS increased significantly, reaching 3.20 ± 0.73 cells /200 μπι at 60 minutes, a four-fold increase compared to the control group. The increase in the number of leukocytes induced by LPS resulted in a significant improvement after administration of R1, reaching 1.20 ± 0.37 cells / 200 μπι at 60 minutes. The percentage of mast cells that were degranulated along the mesenteric venules observed 60 minutes after LPS injection, the percentage of degranulated mast cells in the control group was 19.8 ± 8.1%, representing spontaneous mast cell degranulation. The number of degranulated mast cells that rose after 60 minutes of LPS injection increased to 49.4 ± 7.3%. give Rbl and Rl significantly inhibited LPS-induced degranulation of mast cells (13.9 ± 3.8% and 23.8 ± 4.1%, respectively, at the end of the observation period). Rgl also showed a certain inhibition of LPS-induced degranulation of mast cells (observed end point reached 30.1 ± 7.4%).

The concentrations of plasma cytokines, TNF-α and IL-6, were 24.51 ± 5.57 and 22.72 ± 8.67 ng/L, respectively, in the control group. The two cytokines measured rapidly rose after LPS stimulation, reaching 493.97 ± 192.29 and 741.53 ± 126.86 ng/L, respectively. Administration of Rbl, Rgl and Rl significantly inhibited LPS-induced IL-6 production.

The proportion of cells positive for both CDllb and CD18 was essentially the same under all test conditions. However, if the expression of these adhesion molecules is represented by fluorescence intensity, there is a significant difference. The fluorescence intensity of CD1 lb increased rapidly after LPS stimulation (247.9 ± 6.9, 1.8 fold compared to the control), and Rbl and R1 significantly inhibited LPS-induced increase in CD1 lb expression and showed a dose-dependent state. Even more pronounced effects were observed with R1, and the expression of neutrophil CD18 was similar to that of CDl lb. After LPS stimulation, fluorescence intensity increased rapidly, and LPS-induced CD18 expression was significantly reduced by Rbl and showed a dose-dependent relationship (176.8 ± 10.3, 1.0 mg/ml and 138.9 ± 12.7, 2.0 mg/ml) using Rl pre- Treatment resulted in a more pronounced reduction in LPS-induced CD18 expression (119.6 ± 7.9, 0.2 mg/ml), although there was no dose-dependent relationship like CD1 lb-like.

The above experimental results show that injection of LPS can reduce the movement speed of red blood cells, but Rbl, Rgl and Rl can attenuate this effect of LPS. Injection of LPS can lead to adhesion of leukocytes to the wall, degranulation of mast cells, and release of cytokines. Rbl, Rgl and Rl can reduce the number of adherent cells, inhibit mast cell degranulation and inhibit the increase of cytokine levels. These results provide a new alternative to the clinical treatment of LPS-induced sepsis. detailed description:

The invention is further illustrated by the following examples, which are not intended to limit the invention.

Example 1

Tablet

[Prescription] Rbl 100g Microcrystalline cellulose 50g Microsilica gel 3g Magnesium stearate 1. 5g [Preparation method] Take the original and auxiliary materials through 100 mesh sieves separately; take notoginsenoside, microcrystalline cellulose, mix well, use 60% ethanol as the binder to make soft materials, pass 20 mesh sieve particles, 60'C Dry, take out, pass through 30 mesh sieve, add micro-silica gel and magnesium stearate, mix and compress, and make 1000 pieces.

Example 2

[Prescription] Rgl 75g microcrystalline cellulose 37g micro powder silica gel 2. 3g magnesium stearate l . l g

[Preparation method] Take the original and auxiliary materials through 100 mesh sieves separately; take notoginsenoside, calcium sulfate, microcrystalline cellulose, mix well, use 60% ethanol as the binder to make soft materials, and filter the particles through 20 mesh. 60'C is dried, taken out, sieved through 30 mesh, added with appropriate amount of micro-silica gel and magnesium stearate, mixed, and compressed into 1000 pieces.

Example 3

[Prescription] Rl 133g Microcrystalline Cellulose 66g Microsilica Gel 4g

Magnesium stearate 2g

[Preparation method] Take the original and auxiliary materials through 100 mesh sieves separately; take notoginsenoside, calcium sulfate, microcrystalline cellulose, mix well, use 60% ethanol as the binder to make soft materials, pass 20 mesh sieve particles, Dry at 60 ° C, take out, sift through 30 mesh, add appropriate amount of micro-silica gel and magnesium stearate, mix and compress, and make 1000 pieces.

Example 4

Capsule

Take Rbl, add appropriate amount of starch, magnesium stearate and other accessories, granulate, whole, and put into the No. 1 capsule, that is. Example 5

Oral solution

Take Rgl, add appropriate amount of sucrose, preservative, add water to 1000ml, and dispense into 10ml - branch, that is, get oral liquid.

Example 6

Granule

Take Rl, add appropriate amount of dextrin, stevia, dry granulation, whole granules, and dispense.

Example 7 u injection

Rbl 150g is dissolved in water, and sodium chloride and ethyl p-hydroxybenzoate are dissolved in water, mixed, and adjusted to pH. The water for injection is diluted to 1000 ml, filtered through a hollow fiber membrane, filled, and sterilized. Example 8

Application example, treatment of sepsis

Typical cases

Patient X X, male, 55 years old, staff member, has headache, nausea, vomiting, bloating, abdominal pain, general discomfort, muscle and joint pain, etc., diagnosed as sepsis

Start injection of Rbl injection, size 0. 05g / support, 2 times a day, three times a day. After half a month, the symptoms were obviously improved. There were no adverse reactions during the medication.

Claims

Rights request

1. The use of Panax notoginseng saponins in the preparation of a medicament for treating sepsis.

2. Use according to claim 1, characterized in that the notoginsenoside component is Rbl.

3. The use of claim 1, wherein the notoginsenoside component is Rgl.

4. The use according to claim 1, characterized in that the notoginsenoside component is R1.

The use according to claim 1, characterized in that the sepsis is an acute systemic infection caused by the incursion of pathogenic bacteria or conditional pathogens into the blood circulation to produce toxins and other metabolites.

6. Use according to claim 1, characterized in that the sepsis triggers a microcirculatory disorder which is a systemic inflammatory response caused by excessive release of LPS.

7. Use according to claim 1, characterized in that said application is that Rbl, Rgl and R1 reduce the number of adherent cells and attenuate the decrease in the rate of movement of red blood cells caused by LPS.

8. Use according to claim 1, characterized in that the application is that Rbl, Rgl and R1 inhibit mast cell degranulation and inhibit the increase of cytokine levels.

The use according to claim 1, wherein the notoginsenoside component is a pharmaceutical preparation composition containing a notoginsenoside component.

The use according to claim 9, characterized in that the pharmaceutical preparation composition has Rbl, Rgl or/and R1 as a pharmaceutically active ingredient and a pharmaceutically acceptable carrier, wherein the weight of the notoginsenoside in the preparation The percentage is from 0.1 to 99.9%, the balance being a pharmaceutically acceptable carrier.