CN101327236B - Use of pseudo-ginseng saponin component for treating septicemia - Google Patents

Use of pseudo-ginseng saponin component for treating septicemia Download PDF

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CN101327236B
CN101327236B CN2007101112786A CN200710111278A CN101327236B CN 101327236 B CN101327236 B CN 101327236B CN 2007101112786 A CN2007101112786 A CN 2007101112786A CN 200710111278 A CN200710111278 A CN 200710111278A CN 101327236 B CN101327236 B CN 101327236B
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lps
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韩晶岩
孙凯
郭俊
王传社
刘育英
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Tasly Pharmaceutical Group Co Ltd
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Abstract

The invention relates to an application of notoginsenoside for treating septicemia, particularly relates to ginsenoside Rb1 which is the effective component of notoginsenoside, and an application of ginsenoside Rb1 and notoginsenoside R1 in treating and/or preventing septicemia.

Description

The application of pseudo-ginseng saponin component for treating septicemia
Technical field:
The present invention relates to the new purposes of tcm product, particularly arasaponin composition Rb1, Rg1 and R1 are treating and/or preventing the application of septicemia.
Background technology:
(Panax notoginseng PN) is the root of Chinese Chinese medicine Radix Notoginseng (Burk) F.H.Chen after air-dry to Radix Notoginseng.Arasaponin (is also referred to as: Radix Notoginseng total arasaponins, English name: Panax Notoginsenosides, PNS) be the effective constituent that mainly contains of Radix Notoginseng, form by the different Saponin of kind surplus 30, it is diseases related to be widely used in the treatment microcirculation disturbance clinically, as cardiovascular disease regulating liver-QI dysfunction.
Radix Notoginseng total arasaponins has been listed state-promulgated pharmacopoeia in, and the corresponding preparations launch is arranged, as: panax notoginseng saponins for injection.The ginsenoside Rb1 is called for short the Rb1[English name] Ginsenoside Rb1[chemical name] β-D-Glucopyranoside,
The oxy of (3 β, 12 β)-20-[(6-O-β-D-glucopyranosyl-β-D-glucopyranosyl)]-12-hydroxydammar-24-3n-3yl 2-O β-D-glucopyranosyl-[molecular formula] the C54H92023[molecular weight] 1109.26
Ginsenoside Rg1, be called for short Rg1, [English name] Ginsenoside Rg1[another name] panaxoside A, Sanchinoside Cl[chemical name] β-D-Glucopyranoside, (3 β, 6 α, 12 β)-3,12-dihydroxydammar-24-ene-6, the 20-diylbis-[molecular formula] C42H72014o2H2O.Arasaponin R1 is called for short R1, [English name] Notoginsenoside R1
These three kinds of Saponins are extensively applied to the main active ingredient of the Radix Notoginseng with function of promoting blood circulation to disperse blood clots of Chinese medicine.Septicemia is that pathogenic bacterium or conditioned pathogen are invaded growth and breeding in the blood circulation, produce toxin and the caused acute systemic infection of other metabolites, be feature with shiver with cold, hyperpyrexia, erythra, arthralgia and hepatosplenomegaly clinically, part can have septic shock and migrate the sexually transmitted disease (STD) kitchen range.The reason that causes septicemia has:
One, human factors:
1. when mucocutaneous when breakage being arranged or suppurative inflammation taking place, antibacterial is then easily in the intrusive body.
2. the immunoreation of human body can be divided into two kinds of nonspecific immune reaction and specific immune responses, and the latter can be divided into cellular immunization and humoral immunization two aspects again.When body's immunity descends, can not give full play to the effect that it engulfs kill bacteria, even the invading bacteria amount is less, pathogenicity can not cause septicemia by force yet.
3. the caused nosocomial infection of conditioned pathogen also increases gradually.
Two, antibacterial factor:
Main relevant with virulence and the quantity of pathogen.Virulence pathogenic bacterium strong or that quantity is many enter body, cause that the probability of septicemia is bigger.
The cardinal symptom of septicemia is:
1. former inflammation disease: the caused former inflammation disease of various pathogen is relevant with its distribution position at human body.The characteristics of former inflammation disease be partial red, swollen, hot, the pain and dysfunction.
2. toxemia synptom: how hurried onset is.Often have shiver with cold, hyperpyrexia, heating mostly be remittent fever and or intermittent fever, also can be continued fever, irregular fever and double quotidian fever, due to latter's polyphyly gram-negative bacteria septicemia.Heating is simultaneously with in various degree toxemia synptom, as headache, feel sick, vomiting, abdominal distention, stomachache, the whole body are uncomfortable, muscle and arthralgia etc.
3. erythra: see the part patient, see morely, be distributed in trunk, extremity, eye conjunctiva, oral mucosa etc. more and locate so that petechia is the most, few in number.
4. joint symptom: can occur that big joint is red, swollen, hot, pain and limitation of activity, even concurrent arthroedema, empyema, be more common in the course of disease of septicemia such as gram positive coccus, meningococcus, Bacillus alcaligenes.
5. septic shock: see 1/5～1/3 septic patient approximately, show as dysphoria, pulse is speed carefully, extreme cold of the limbs, and piebald skin, hypourocrinia and blood pressure drops etc., and DIC can take place are due to the serious toxemia.
6. hepatosplenomegaly: general silght enlargement only.
According to research, lipopolysaccharide (LPS) is one of constituent of gram-positive bacterium cell wall, has caused the people's septicemia that gram-positive bacterium causes and the multiple performance of septic shock.The microcirculation that takes place during septicemia barrier is the systemic inflammatory responses that the excessive release by LPS causes, mediated by many activated factors, these factors such as adhesion molecule, active oxygen composition (ROS), and inflammation precursor medium such as tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and interferon-γ (INF-γ).The organ dysfunction that the microcirculation disturbance that LPS induces generation takes place during to septicemia plays pivotal role, leukocyte is along endothelium rotation and adhere to blood vessel endothelium in this process, and blood vessel wall produces hydrogen peroxide and mast cell degranulation all is the committed step of bibliographical information.
The main clinical treatment method of current septicemia comprises the capacity recovery, the utilization of catecholamine and antibiotic compensation treatment.These methods can increase septicemia patient's survival rate, but can cause the decline of blood sugar increasing and blood pressure and blood calcium, the concurrent blood that bears clinically.The clinical research suggestion adopts anti-TNF-α treatment helpful to the treatment of septicemia.Yet, nearest studies show that this method can not cure septicemia and can increase serious septicemia patient's mortality rate.Thereby, in the septicemia that LPS causes, seek the method that prevents serious microcirculation barrier and remain a challenge.
The inventor is carrying out in the research process arasaponin, find that three kinds of Saponin Rb1, Rg1 in the Radix Notoginseng and R1 can weaken the erythrocytic movement velocity that injection LPS causes and reduce, Rb1, Rg1 and R1 can reduce the number of attached cell, the rising that suppresses mast cell degranulation and suppress cytokine levels.In vitro tests utilization flow cytometer has confirmed that further Rb1, Rg1 can suppress the raising that CD11b/CD18 expresses in the neutrophilic granulocyte that LPS causes significantly, and Rb1, Rg1 can suppress LPS and stimulate and cause H 2O 2From the granulocytic release of neutrality.Above result shows, three kinds of Saponin Rb1, Rg1 and R1 can improve the inductive microcirculation disturbance of LPS.Septicemia there is the obvious treatment effect.
Summary of the invention:
The invention provides the new therapeutic use of a kind of pseudo-ginseng saponin component.Described new therapeutic use is the septicemia that causes because of microcirculation disturbance with pseudo-ginseng saponin component for treating.
For this reason, the invention provides a kind of new medicine use, promptly prepare a kind of medicine that treats and/or prevents septicemia with pseudo-ginseng saponin component.
Pseudo-ginseng saponin component of the present invention is selected from the ginsenoside Rb1, ginsenoside Rg1 and arasaponin R1.
Three kinds of saponin component ginsenoside Rb1s of the present invention, ginsenoside Rg1 and arasaponin R1 are prior aries, can buy from the market, also can meet medicinal standard and get final product according to prior art for preparing.Preferred purity〉50%, more preferably purity〉90%, purity most preferably〉98%.
The medicine of preparation of the present invention is the pharmaceutical preparations composition that is prepared into as active constituents of medicine with above-mentioned pseudo-ginseng saponin component.
Pharmaceutical preparations composition of the present invention can contain the medicine acceptable carrier as required, and wherein arasaponin is as active constituents of medicine, and its shared percentage by weight in preparation can be 0.1-99.9%, and all the other are the medicine acceptable carrier.Pharmaceutical preparations composition of the present invention exists with unit dosage form, and described unit dosage form is meant the unit of preparation, as every of tablet, and capsular every capsules, every bottle of oral liquid, every bag of granule, every of injection etc.
Pharmaceutical preparations composition of the present invention can be any pharmaceutically useful dosage form, and these dosage forms comprise: tablet, sugar coated tablet, film coated tablet, enteric coated tablet, capsule, hard capsule, soft capsule, oral liquid, suck agent, granule, electuary, pill, powder, unguentum, sublimed preparation, suspensoid, powder, solution, injection, suppository, ointment, plaster, cream, spray, drop, patch.
Chinese medicine preparation of the present invention, the preparation of its oral administration can contain excipient commonly used, such as binding agent, filler, diluent, tablet agent, lubricant, disintegrating agent, coloring agent, flavoring agent and wetting agent, can carry out coating to tablet in case of necessity.
The filler that is suitable for comprises cellulose, mannitol, lactose and other similar filler.Suitable disintegrating agent comprises starch, polyvinylpyrrolidone and starch derivatives, for example sodium starch glycollate.Suitable lubricant comprises, for example magnesium stearate.The acceptable wetting agent of appropriate drug comprises sodium lauryl sulphate.
Can fill by mixing, the method that tabletting etc. are commonly used prepares solid oral composition.Mix repeatedly active substance is distributed in those compositionss of a large amount of filleies of whole use.
The form of oral liquid for example can be aqueous or oily suspensions, solution, Emulsion, syrup or elixir, perhaps can be a kind of available water before use or other suitable composite dry products of carrier.This liquid preparation can contain conventional additive, such as suspending agent, for example sorbitol, syrup, methylcellulose, gelatin, hydroxyethyl-cellulose, carboxymethyl cellulose, aluminium stearate gel or hydrogenation edible fat, emulsifying agent, for example lecithin, anhydro sorbitol monooleate or arabic gum; Non-aqueous carrier (they can comprise edible oil), for example almond oil, fractionated coconut oil, such as oily ester, propylene glycol or the ethanol of the ester of glycerol; Antiseptic, for example para hydroxybenzene methyl ester or propyl p-hydroxybenzoate or sorbic acid, and if desired, can contain conventional flavouring agent or coloring agent.
For injection, the liquid unit dosage forms of preparation contains active substance of the present invention and sterile carrier.According to carrier and concentration, this chemical compound can be suspended or dissolving.The preparation of solution is normally by being dissolved in active substance in a kind of carrier filter-sterilized before it is packed into a kind of suitable bottle or ampoule, sealing then.For example a kind of local anesthetic of adjuvant, antiseptic and buffer agent also can be dissolved in this carrier.In order to improve its stability, can be after the bottle of packing into that this compositions is freezing, and under vacuum, water is removed.
Chinese medicine preparation of the present invention, when being prepared into medicament, optionally add suitable medicine acceptable carrier, described medicine acceptable carrier is selected from: mannitol, sorbitol, sodium pyrosulfite, sodium sulfite, sodium thiosulfate, cysteine hydrochloride, TGA, methionine, vitamin C, the EDTA disodium, EDTA calcium sodium, the alkali-metal carbonate of monovalence, acetate, phosphate or its aqueous solution, hydrochloric acid, acetic acid, sulphuric acid, phosphoric acid, aminoacid, sodium chloride, potassium chloride, sodium lactate, xylitol, maltose, glucose, fructose, dextran, glycine, starch, sucrose, lactose, mannitol, silicon derivative, cellulose and derivant thereof, alginate, gelatin, polyvinylpyrrolidone, glycerol, soil temperature 80, agar, calcium carbonate, calcium bicarbonate, surfactant, Polyethylene Glycol, cyclodextrin, β-cyclodextrin, the phospholipid material, Kaolin, Pulvis Talci, calcium stearate, magnesium stearate etc.
Preparation of the present invention is determined usage and dosage according to patient's situation in use, but obeys every day three times, each 1-20 agent, as: 1-20 bag or grain or sheet, every dose of 1mg-1000mg.
Therapeutic use of the present invention experimental results show that by following:
Material and method
Reagent and experimental animal
Rb1 and Rg1 are available from Nat'l Pharmaceutical ﹠ Biological Products Control Institute, and R1 is available from Kunming Feng-Shan-Jian pharmaceutical companies.All Saponins all use the affinity chromatography purification.
Male sprague-dawley (SD) rat, body weight obtains from Department Of Medicine, Peking University's animal center between 200～250 grams.The credit number of this batch animal is SCXK2002-0001.Adopt code test chamber Mus feedstuff (Department Of Medicine, Peking University's animal center) to feed before the test 72 hours, keep body temperature at 24 ± 1 ℃, relative humidity is 50 ± 1%, and all experimental animals are accepted 12/12-light/dark cycle.Before the test animal fasting be can't help water 12 hours.The guide of abideing by zooscopy committee of Peking University carries out the animal processing.
Mesenteric mesaraic microcirculation
The utilization urethane is anaesthetized (1.25mg/kg body weight) to the test rat, adopts median incision cut-up test rat abdominal cavity (long 20-30mm).Carry out femoral vein and internal jugular vein intubate test injection medicine.
Careful separation goes out mesenteric mesaraic ileocecus (zone of mesentery afterbody 10-15cm), and mesentery is taken out and is tiled on the transparent plastic platform from intraperitoneal.Continue 37 ℃ of normal saline of perfusion by the surface and keep mesenteric mesaraic temperature and humidity.(Germany, Leica DM-IRB) observe the microcirculatory hematodinamics situation of mesentery to adopt the inverted microscope that uses transilluminator.A television camera (Japan, the Jk-TU53H of Toshiba) and images to a color display are installed on microscope, and (J2118A), (China, Malata DVR-R25) writes down all images by DVD player for China, TCL.This research is selected unramified list to prop up and is not had the object of the venule of obvious bending as experimental observation, and between the 50 μ m, length is about 200 μ m at 30 μ m for venular diameter.
Administration
The test rat is divided into five groups, 6 every group at random.Rat for LPS group and matched group, earlier experimental animal is carried out 10 minutes previewing, then continue medication saline (1ml/hr, matched group) by right jugular vein, be dissolved in brinish Saponin (Rb1, Rg1 or R1) (5mg/kg/hr, simple Saponin group), and be dissolved in brinish LPS (2mg/kg/hr, the LPS group), the time is 60 minutes.For three test group, organizing identical method through right jugular vein employing with LPS for every group increases injection Saponin Rb1 (Rb1+LPS group), Rg1 (Rg1+LPS group) or R1 (R1+LPS group) more respectively.20 minutes start injection Saponins before injection LPS, the continuous injection Saponin is until observe end.Every group of Saponin that uses all is dissolved in saline in advance then with the dosed administration of 5mg/kg/hr in three groups.Continue to observe and write down the microcirculatory hemodynamic state of rat mesentery of each group.
The mensuration of blood vessel diameter
(Media Cybernetic, when USA) software is respectively to lps injection (0 minute), and the image that records when 20 minutes, 40 minutes and 60 minutes after the lps injection carries out the mensuration of blood vessel diameter to use Image-Pro Plus5.0.Same position is measured blood vessel diameter and is got its meansigma methods then three times.
The mensuration of erythrocyte speed
The CCD of monitor is converted to high speed video photographic system (FASTCAM-ultima APX, photron, Japan) with erythrocytic speed in the speed record venule of 2000 frames/s, afterwards the high speed image that stores with the speed of 25 frames/s from new broadcast.When lps injection (0 minute) respectively, and after the lps injection 20 minutes, 40 minutes and use Image-Pro Plus5.0 software 60 minutes the time (Media Cybernetic USA) measures erythrocytic speed in the venule.
Shear rate is measured
The shear rate of using formula SR=2.12 * 8 * [V/D] estimation venule blood vessel wall, the D here represents the blood vessel average diameter, and V is erythrocytic Mean Speed, the 2.12 experience correction factors for blood capillary velocity profile in the body that measures.
The mensuration of leukocyte adhesion
Thereby come the leukocytic dynamic behavior of retrospective analysis to estimate the adhesion of leukocyte to blood vessel wall by the image that replay records.In this research, be defined as and adhere to the homonymy blood vessel wall time and surpass the cell of 10s adhering to leukocyte.20 minutes, 40 minutes and 60 minutes after (0 minute) and the lps injection when lps injection respectively, the number of the interior attached cell of the blood vessel segment that the picture count that records by picked at random is certain (diameter 30～50 μ m, length is 200 μ m).
The mensuration of leucocyte migration
The leukocyte image that records when 20 minutes, 40 minutes and 60 minutes after (0 minute) and the lps injection when reviewing lps injection is estimated the leukocyte number that moves.The leukocyte of migration is defined as the leukocyte number around the venule of certain-length, is that the blood vessel of 200 μ m is measured the leukocyte number (diameter 30～50 μ m) in the surrounding tissue (diameter 30～50 μ m) by selecting the image of record at random and selecting length.The mensuration of mast cell degranulation
Injection LPS 60 minutes afterwards, the toluidine blue perfused tissue with 0.1% 1 minute is bleached with saline then.The cell that the cell endoparticle is discharged into surrounding tissue is considered to degranulated mastocyte, and the cell number in counting 20 * object lens microscope circular field of view is determined the number of this class cell.Select blood capillary system 5 visuals field on every side to estimate each mesentery window.Record takes off granule and non-degranulated mastocyte number, calculates the ratio of taking off the granule mastocyte then.
The mensuration of cytokine
Observed after the mesentery microcirculation, gathered the blood preparation of each experimental animal through the ventral aorta approach and carry out anticoagulant heparin (20unit/ml blood).Pass through centrifugal separation plasma then.50 μ l blood plasma and standard substance and 50 μ l attract pearl to hatch altogether 1 hour under room temperature, mix with the TNF-α and the IL-6 detection antibody of 50 μ l PE labellings then, hatch altogether under room temperature and form the sandwich complex in 2 hours.After the hatching, every pipe adds the cleaning buffer solution (BD Biosciences Pharmingen, the U.S.) of 1ml altogether, and utilization flow cytometer (FACS Calibur, B.D.Co, the U.S.) detects average fluorescent strength.Adopt BD Cytometric Bead Array analysis software to carry out data analysis.
The external test that CD11b/CD18 expresses
Other one group of (n=6) SD rat is carried out the external test that CD11b/CD18 expresses.Rat fasting before test be can't help water 12 hours.Adopt urethane that rat is anaesthetized (1.25mg/ per kilogram of body weight, intramuscular injection).Abdominal aortic blood from each experimental animal, use anticoagulant heparin (20 units/every ml blood) then, prepare 20 duplicate samples according to following requirements then: contrast, Rb1 (0.2mg/ml), Rg1 (0.2mg/ml), R1 (0.2mg/ml), Rb1 (1.0mg/ml), Rg1 (1.0mg/ml), R1 (1.0mg/ml), Rb1 (2.0mg/ml), Rg1 (2.0mg/ml), R1 (2.0mg/ml), LPS, Rb1+LPS (0.2mg/ml), Rg1+LPS (0.2mg/ml), R1+LPS (0.2mg/ml), Rb1+LPS (1.0mg/ml), Rg1+LPS (1.0mg/ml), R1+LPS (1.0mg/ml), Rb1+LPS (2.0mg/ml), Rg1+LPS (2.0mg/ml) and R1+LPS (2.0mg/ml).In the middle of the sample of each part preparation, add the blood of 0.2ml.Matched group is the saline of 8 μ l.Single Saponin group is made of a kind of in three kinds of Saponins of 4 μ l saline and 4 μ l, and ultimate density is seen within the above round parentheses.The LPS sample is made of 4 μ l saline and 4 μ l LPS, finally makes the concentration of LPS reach 2 μ g/ml.Other all samples are made of a certain in three kinds of Saponins of 4 μ l LPS and 4 μ l, finally make the concentration of LPS reach 2 μ g/ml, and the ultimate density of each test Saponin is seen in the above round parentheses (Sun et al., 2006).Biased sample was preserved 2 hours under 37 ℃, hatched altogether under room temperature 20 minutes with the anti-CD11b antibody or the anti-CD18 antibody (BD Biosciences Pharmingen, the U.S.) of 1ugFITC-labelling then.Adopt hemolysin dissolved cell (BD Biosciences Immunocytometer Systems, the U.S.), clean twice with PBS then.Use flow cytometer (FACS Calibur, B.D.Co, the U.S.) to measure average fluorescent strength.Adopt the FSC-SSC scatterplot to select neutrophilic granulocyte.Every duplicate samples is measured 5,000 neutrophilic granulocyte number, calculates average fluorescence intensity then.
H in the external neutrophilic granulocyte cell 2O 2With O 2 -The analysis of output
Adopt anticoagulant heparin then from abdominal aortic blood.Use the mono-poly dissolve medium to separate neutrophilic granulocyte (Ting and Morris, 1971) according to the method for Ting report.According to improved Shen method (Shen etal., 1998), utilization RPMI1640 medium prepares cell suspension and uses flow cytometer (BD company, USA) mensuration cell hydrogen peroxide (H then then 2O 2) and superoxide anion ( O 2 -) output.Briefly process is, neutrophilic granulocyte is at 37 ℃ of following and 0.2mM2 ', and 7 '-dichlorofluorescein diacetate (DCFH-DA) was hatched 5 minutes altogether, hatched altogether 15 minutes with the dihydro bromination second pyridine (HE) of 0.1mM once more.Use Rb1 (1.0mg/ml) then separately, Rg1 (1.0mg/ml) or R1 (1.0mg/ml) handle cell, perhaps use LPS (2 μ g/ml) to handle simultaneously.After hatching 20 minutes altogether with ice cube, with flow cytometer by measuring 2 ', 7 '-dichlorofluorescein (DCF) emission 525nm fluorescence and the fluorescence of the 590nm of ethidium bromide (EB) emission measure H 2O 2With O 2 -Content.
The mensuration of blood plasma alkali phosphatase
After having observed mesenteric mesaraic microcirculation, use anticoagulant heparin (the every ml blood of 20unit/) then from the abdominal aortic blood of every experimental animal.Go out blood plasma by centrifugalize.(7170A, Hitachi Japan), adopt alkaline phosphatase enzyme reagent kit (Olympus, US) concentration of detection blood plasma neutral and alkali phosphatase to the utilization automatic biochemistry analyzer.
Statistics
Adopt variance analysis and F-check to carry out statistical analysis.Numerical value is represented with meansigma methods ± S.E (N=6).The P value thinks that less than 0.05 statistical remarkable meaning is arranged.
The result
Between the diameter of the mesenteric venule that 0 time point and other times point detect, there is not significant difference after the lps injection.Each experimental condition mesenterium ventrale venule erythrocyte speed over time, erythrocyte speed did not change in 60 minutes observation period in the matched group venule.On the contrary, the carrying out property reduction after lps injection 40 minutes of the venular erythrocyte speed of LPS group rat mesentery, and be lower than 0 time point and matched group (p＜0.05) significantly.The injection after 60 minutes, the erythrocytic speed that measures is 1.30 ± 0.11mm/ second, is 70% of 0 time value.Rb1 can change the reduction of the inductive mesenteric venule erythrocyte of rat LPS speed on to a certain degree after Rg1 or R1 handle.
The rat mesentery venule red blood cell adhesion situation of process in time is, at 0 time point, the leukocyte number that adheres to the venule wall does not have significant difference between these groups, in the observation end of term of matched group, adherent leukocyte number only has slight 1.72 ± 0.71cells/200 μ m that risen to.On the contrary, the value of LPS group then shows the linear growth relation of tangible time and number, a high value the when 5.00 ± 0.83cells/200 μ m after lps injection 20 minutes the time has risen to 60 minutes has reached 12.72 ± 1.41cells/200 μ m, increases than the value of 0 time point to have surpassed 15 times.Give Rb1, all cause the inductive leukocyte of LPS that the venule wall is adhered to after Rg1 or the R1 and obtained improvement, though for each Saponin along with the process of time has some variations, the inhibitory action of R1 appears at 20 minutes after the lps injection, the inhibitory action of Rb1 appears at 40 minutes after the lps injection, and the inhibitory action of Rg1 appears at 60 minutes after the lps injection.
Leukocyte is along with the situation that time course moves out of the mesenteric venule wall is, any situation 0 time point does not all detect the tangible leukocyte that moves out of blood vessel wall.For matched group, almost all do not observe the leukocyte that moves out of blood vessel wall in the time period that whole sight is looked into.On the contrary, the number of WBC phenomenal growth of moving out of wall of vein after the injection LPS has reached 3.20 ± 0.73cells/200 μ m in 60 minutes, and comparing with matched group has increased by four times.The inductive migration leukocyte count of LPS purpose is increased in and gives to have obtained significant improvement after the R1, has reached 1.20 ± 0.37cells/200 μ m in 60 minutes.Observed percentage ratio along the degranulated mastocyte of mesenteric venule after 60 minutes after injection LPS, the percentage ratio of the degranulated mastocyte of matched group is 19.8 ± 8.1%, has represented idiopathic mast cell degranulation.Inject the number that degranulated mastocyte took place afterwards in LPS60 minute and risen to 49.4 ± 7.3%.Give Rb1 and R1 and can suppress the inductive mast cell degranulation of LPS (terminal point in the observation period has reached 13.9 ± 3.8% and 23.8 ± 4.1% respectively) significantly.Rg1 has also shown the effect (observe terminal point and reached 30.1 ± 7.4%) of certain inductive mast cell degranulation of inhibition LPS.
The concentration situation of plasma cell factor TNF-α and IL-6 is, at matched group, the concentration of TNF-α and IL-6 is respectively 24.51 ± 5.57 and 22.72 ± 8.67ng/L.The rising rapidly after LPS stimulates of two cytokines measuring has reached 493.97 ± 192.29 and 741.53 ± 126.86ng/L respectively.Give Rb1, Rg1 and R1 can significantly suppress the generation of the inductive IL-6 of LPS.
CD11b substantially is consistent under all experimental conditions with the positive cell proportion of CD18.Yet,, evident difference is arranged if represent the expression of these adhesion molecules with fluorescence intensity.The fluorescence intensity of CD11b raise (be compared to contrast and increased by 247.9 ± 6.9,1.8 times) rapidly after LPS stimulated, and Rb1 and R1 can significantly suppress the raising of the inductive CD11b expression of LPS and show dose-dependent state.Use R1 even observed more tangible effect, the expression of neutrophilic granulocyte CD18 is similar to the expression of CD11b.After LPS stimulated, fluorescence intensity increased rapidly, the inductive CD18 of LPS express can be reduced by Rb1 significantly and present dose-dependent relation (176.8 ± 10.3,1.0mg/ml and 138.9 ± 12.7,2.0mg/ml).Utilization R1 pretreatment cause more significantly reducing the effect that the inductive CD18 of LPS expresses (119.6 ± 7.9,0.2mg/ml), though do not show dose-dependent relation as CD11b.
Above experimental result shows that injection LPS can reduce erythrocytic movement velocity, but Rb1, Rg1 and R1 can weaken this effect of LPS.Can cause the release of leukocyte behind the injection LPS to adhesion, mast cell degranulation and the cytokine of tube wall.And Rb1, Rg1 and R1 can reduce the number of attached cell, the rising that suppresses mast cell degranulation and suppress cytokine levels.These results provide a new alternative method for the clinical treatment of the inductive septicemia of LPS.
The specific embodiment:
Further specify the present invention by the following examples, but not as limitation of the present invention.
Embodiment 1
Tablet
[prescription] Rb1100g microcrystalline Cellulose 50g micropowder silica gel 3g magnesium stearate 1.5g
[method for making] got former, adjuvant and crossed 100 mesh sieves respectively; Get arasaponin, microcrystalline Cellulose, mixing in right amount as binding agent system soft material, is crossed 20 mesh sieve system granules with 60% ethanol, and 60 ℃ of dryings are taken out, and cross 30 mesh sieve granulate, add micropowder silica gel and magnesium stearate, mixing, and tabletting is made 1000, promptly.
Embodiment 2
[prescription] Rg175g microcrystalline Cellulose 37g micropowder silica gel 2.3g magnesium stearate 1.1g
[method for making] got former, adjuvant and crossed 100 mesh sieves respectively; Get arasaponin, calcium sulfate, microcrystalline Cellulose, mixing in right amount as binding agent system soft material, is crossed 20 mesh sieve system granules with 60% ethanol, 60 ℃ of dryings are taken out, and cross 30 mesh sieve granulate, add an amount of micropowder silica gel and magnesium stearate, mixing, tabletting is made 1000, promptly.
Embodiment 3
[prescription] R1133g microcrystalline Cellulose 66g micropowder silica gel 4g magnesium stearate 2g
[method for making] got former, adjuvant and crossed 100 mesh sieves respectively; Get arasaponin, calcium sulfate, microcrystalline Cellulose, mixing in right amount as binding agent system soft material, is crossed 20 mesh sieve system granules with 60% ethanol, 60 ℃ of dryings are taken out, and cross 30 mesh sieve granulate, add an amount of micropowder silica gel and magnesium stearate, mixing, tabletting is made 1000, promptly.
Embodiment 4
Capsule
Get Rb1, add appropriate amount of starch, adjuvants such as magnesium stearate are granulated, granulate, and the capsule of packing into No. 1, promptly.
Embodiment 5
Oral liquid
Get Rg1, add an amount of sucrose, antiseptic adds water to 1000ml, is distributed into one of 10ml, promptly gets oral liquid.
Embodiment 6
Granule
Get R1, add an amount of dextrin, steviosin, dry granulation, granulate, packing, promptly.
Embodiment 7
Injection
Rb1150g is dissolved in water, and sodium chloride, ethylparaben add the hot water dissolving, mixing, adjust pH in addition.Water for injection is diluted to 1000ml, filters with hollow-fibre membrane, and fill, sterilization, promptly.
Embodiment 8
Application example, the treatment of septicemia
Model case
The patient opens * *, the male, 55 years old, the office worker had headache, feels sick, vomiting, abdominal distention, stomachache, the whole body are uncomfortable, muscle and arthralgia etc., are septicemia after diagnosing
Start injection Rb1 injection, specification 0.05g/ props up, each 2 of every day three times.Subjective symptoms takes an evident turn for the better behind the first quarter moon.No any untoward reaction during the medication.

Claims (6)

1. the application of pseudo-ginseng saponin component Rb1 in a kind of medicine for the treatment of septicemia of preparation.
2. the application of pseudo-ginseng saponin component R1 in a kind of medicine for the treatment of septicemia of preparation.
3. claim 1 or 2 application is characterized in that, described septicemia is that pathogenic bacterium or conditioned pathogen are invaded growth and breeding in the blood circulation, produces toxin and the caused acute systemic infection of other metabolites.
4. claim 1 or 2 application is characterized in that, described septicemia causes microcirculation disturbance, and this microcirculation disturbance is the systemic inflammatory responses that the excessive release by LPS causes.
5. claim 1 or 2 application is characterized in that, described application is the number that Rb1 or R1 reduce attached cell, weaken the erythrocytic movement velocity that LPS causes and reduce.
6. claim 1 or 2 application is characterized in that, described application is the rising that Rb1 or R1 suppress mast cell degranulation and suppress cytokine levels.
CN2007101112786A 2007-06-21 2007-06-21 Use of pseudo-ginseng saponin component for treating septicemia Active CN101327236B (en)

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JP2000191539A (en) * 1998-12-22 2000-07-11 Japan Science & Technology Corp BRAIN CELL OR NERVE CELL-PROTECTING AGENT COMPRISING GINSENOSIDE Rb1
CN1623555A (en) * 2003-12-05 2005-06-08 云南金不换(集团)有限公司 Notoginseng saponin drops
CN100500687C (en) * 2004-08-04 2009-06-17 李明劲 Process for preparing notoginseng triol saponin and use thereof
CN100384429C (en) * 2006-04-07 2008-04-30 黑龙江省珍宝岛制药有限公司哈尔滨分公司 Capsule for treating cardiac and cerebral vascular diseases and its preparing method and application

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毕秀丽,等.人参皂苷类成分对活化的小胶质细胞NO及TNFα释放的不同调节作用.《中国药理通讯》.2004,第21卷(第3期),8. *

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