CN111000919A - Application of ethanol extract of evodia lepta in preparation of medicine for treating sepsis - Google Patents

Application of ethanol extract of evodia lepta in preparation of medicine for treating sepsis Download PDF

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CN111000919A
CN111000919A CN201911332041.XA CN201911332041A CN111000919A CN 111000919 A CN111000919 A CN 111000919A CN 201911332041 A CN201911332041 A CN 201911332041A CN 111000919 A CN111000919 A CN 111000919A
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姚琦
高英
李红日
武春兰
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Guizhou University of Traditional Chinese Medicine
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Abstract

The invention discloses application of an ethanol extract of evodia lepta in preparing a medicament for treating sepsis. Provides a new idea for the research and development of sepsis therapeutic drugs, has rich trifacial bitter resources, is easy to obtain, can drive the full development and utilization of medicinal material resources, and generates related economic benefits.

Description

Application of ethanol extract of evodia lepta in preparation of medicine for treating sepsis
Technical Field
The invention relates to a medicament for treating sepsis, in particular to application of an ethanol extract of evodia lepta in preparing a medicament for treating sepsis.
Background
Clinically, gram-negative (G)－) Severe bacterial infections often lead to the development of Systemic Inflammatory Response Syndrome (SIRS) and eventually to Sepsis (Sepsis), Septic shock (Septic shock) and Multiple Organ Dysfunction Syndrome (MODS). The mortality rate of sepsis is high, wherein the mortality rate of severe sepsis is more 30% -70%, and the sepsis is considered as one of the major diseases seriously threatening the life and health of human beings. International guidelines for treatment of severe sepsis and septic shock recognize that sepsis treats nucleiThe heart is primarily responsible for early infection control and supportive care. Given that sepsis 90% of the pathogens are bacteria, early empirical anti-infective therapy is critical for the prognosis of critically ill patients. The application of a large amount of broad-spectrum and high-efficiency antibiotics provides a powerful weapon for clinically treating bacterial infectious diseases, but the wide application of the antibiotics causes the large increase of drug-resistant strains, and the release of endotoxin (LPS) is caused while bacteria are killed, so that the inflammatory reaction is aggravated. Therefore, it is of great significance to find drugs that effectively antagonize sepsis and to study their mechanisms of action.
The Rutaceae plant of the genus Amaranthus is widely distributed in southern China. It is bitter and cold in nature and has the effects of clearing away heat and toxic material, dispelling wind and removing dampness. Chemical research shows that the trigeminal bitter contains various chemical components such as alkaloid, benzopyran, flavone and the like. Pharmacological research finds that the thin evodia leaf extract has strong antibacterial, anti-inflammatory, antioxidant and analgesic effects. However, until now, no study on the use of trifacial bitter for the treatment of sepsis has been found.
Disclosure of Invention
The invention aims to provide the application of the ethanol extract of the trifoliate bitter in preparing the medicine for treating the sepsis, provides a new idea for the research and development of the medicine for treating the sepsis, is easy to obtain the trifoliate bitter, and can drive the full development and utilization of medicinal material resources.
The technical scheme of the invention is as follows: application of evodia lepta in preparing medicine for treating sepsis is provided.
Application of Foliumet extract in preparing medicine for treating sepsis is provided.
In the application of the ethanol extract of the trifacial bitter in the preparation of the medicine for treating sepsis, the ethanol extract of the trifacial bitter is an ethanol extract of the trifacial bitter.
In the application of the ethanol extract of the trifoliate-bitter in the preparation of the medicine for treating sepsis, the ethanol extract of the trifoliate-bitter is obtained by crushing and sieving the trifoliate-bitter leaves, adding 70-90% ethanol for cold soaking for 3-5 times, filtering, taking filtrate, concentrating the filtrate under reduced pressure at 60-80 ℃, and drying the filtrate at 60-85 ℃ to obtain the ethanol extract of the trifoliate-bitter.
In the application of the ethanol extract of the trifacial bitter in the preparation of the medicine for treating sepsis, the ethanol extract of the trifacial bitter is prepared by crushing and sieving the leaves of the trifacial bitter, adding 80% ethanol for cold soaking for four times, filtering, taking filtrate, carrying out reduced pressure distillation and concentration for 1-2 times at 70 ℃, drying the filtrate on a water bath kettle at 80 ℃, and then drying the filtrate in a reduced pressure oven at 70 ℃ to obtain the ethanol extract of the trifacial bitter.
In the application of the ethanol extract of the trifacial bitter in preparing the medicine for treating sepsis, the weight ratio of the trifacial bitter to the ethanol is 1:5-1:10, and the filtrate is concentrated under reduced pressure to 1/3-1/5 of the original volume.
A medicament for treating sepsis comprises a thin evodia leaf or a thin evodia leaf extract.
A medicine for treating sepsis is mainly prepared from the extract of evodia lepta or evodia lepta.
In the preparation method of the medicine for treating sepsis, the evodia lepta or the evodia lepta extract is combined with the auxiliary materials acceptable in the medicine and processed according to the conventional method to prepare the corresponding medicine.
In the preparation method of the medicine for treating sepsis, the medicine is tablets, capsules, granules, oral liquid or pills.
Compared with the prior art, the invention has the following beneficial effects:
1. as shown in the subsequent experimental evidence, the trifacial bitter extract obviously improves the survival rate of mice with sepsis induced by pathogenic bacteria or Lipopolysaccharide (LPS) and other factors, and the trifacial bitter ethanol extract also has obvious inhibiting effect on inflammatory cytokines generated during sepsis on the levels of animals and cells. Therefore, the invention provides a feasible technical scheme for applying the trigeminal bitter extract to the treatment of the sepsis, and analyzes the possibility of applying the trigeminal bitter extract to the market as a medicament.
2. The experimental result of the invention shows that the tail vein is passedDifferent pathogenic bacteria were injected at a minimum dose of 100% animal deaths (100% MLD). Wherein, the ratio of staphylococcus aureus ATCC29213 to staphylococcus aureus ATCC29213 is 3.3 multiplied by 1010CFU/ml, Klebsiella pneumoniae ATCC10031: 2.0X 1010CFU/ml, Escherichia coli ATCC25922: 1.5X 1010CFU/ml, Pseudomonas aeruginosa ATCC27853: 1.6X 1010CFU/ml) or 40mg/kg LPS, and the trigeminal bitter extract obviously improves the survival rate and survival time of 168h sepsis mice, in addition, enzyme-linked immunosorbent assay (ELISA) is adopted to detect related cell factors in the serum of LPS sepsis-inducing mice, and the trigeminal bitter ethanol extract is found to obviously reduce the expression levels of TNF- α and IL-6 in the serum of LPS sepsis-inducing mice.
Experiments prove that:
1. preparation of ethanol extract of evodia lepta
Collecting Evodia lepta leaf (purchased from Yunnan Lincang good medicine materials planting Limited, and produced in Yunnan Lincang) 8.0kg for dust removal and purification, pulverizing, sieving with 60 mesh sieve, placing in a percolation tank, and cold soaking the medicinal materials with 8 times volume of ethanol (volume ratio of 4:1) for 24 hr to obtain Evodia lepta ethanol extract leaching liquor;
drying the cold soaking solution, storing, and cold soaking the medicinal materials with 8 times volume of ethanol/water solution (volume ratio of 4:1) for 3 days;
drying the second cold soaking solution, storing, and cold soaking the medicinal materials with 8 times volume of ethanol/water solution (volume ratio of 4:1) for 4 days;
draining the cold soaking solution for the third time, storing, and percolating with 8 times volume of ethanol/water solution (volume ratio of 4: 1);
draining percolate, mixing all the above medicinal liquids, and concentrating at 70 deg.C under reduced pressure to obtain total extract of Foliumet ramulus evodiae.
The 32.2kg medicinal extract is obtained by extracting for 4 times and 8.0kg trifoliate bitter leaves.
Weighing 2.0kg of the medicinal materials, soaking, drying the percolate, drying on a water bath kettle at 80 ℃, and drying in a reduced pressure drying oven at 70 ℃ to obtain 150.4g of dry extract.
The extraction rate is (150.4 × 32.2/2)/8000 × 100%: 30.268%.
Through detection and analysis, the ethanol extract of the trifacial bitter contains various components such as flavonoid, alkaloid, phenylpropanoids and the like.
2. Animal experiments
2.1 experiment of therapeutic action of extract of Evodia lepta on mouse sepsis model induced by different pathogenic bacteria
(1) The administration concentration of the medicine of the thin evodia leaf extract (prepared from 1) is as follows: 750mg/kg of ethanol extract of the trifoliate-bitter.
(2) ICR mice (male and female halves), 6-7 weeks old, 20.0 + -2.0 g. The animals were kept in a Specific Pathogen Free (SPF) grade barrier system, placed in dry clean plastic cages, fed with standard laboratory animal feed, and fed with free access water.
(3) Experimental methods animals were randomly grouped by weight, 10 animals per group. Experiments are provided with a blank control group, a pathogenic bacterium control group, a low-dose group of the thin evodia lepta ethanol extract and a high-dose group of the thin evodia lepta ethanol extract.
100% of MLD different pathogenic bacteria are injected into tail vein respectively, and the injection dosage is as follows: staphylococcus aureus ATCC29213: 3.3X 1010CFU/ml, Klebsiella pneumoniae ATCC10031: 2.0X 1010CFU/ml, Escherichia coli ATCC25922: 1.5X 1010CFU/ml, Pseudomonas aeruginosa ATCC27853: 1.6X 1010CFU/ml. The trifacial bitter ethanol extract is directly administrated once by gavage after injection of different pathogenic bacteria, the gavage administration dosage of the low-dose group of the trifacial bitter ethanol extract is 750mg/kg, and the gavage administration dosage of the high-dose group of the trifacial bitter ethanol extract is 1500 mg/kg. The blank control group and the pathogenic bacteria group were given distilled water at 0.2ml/20g simultaneously.
(4) The status and survival of the mice were recorded every 8 hours of observation and the survival rate and survival time of 168h of the mice were calculated.
(5) And the results obtained
TABLE 1 Effect of ethanol extract of Evodia trifoliata on survival and survival time of Staphylococcus aureus ATCC29213 induced septic mice 168h (S)
Figure BDA0002329893070000051
n＝10)
Figure BDA0002329893070000052
Compared with the blank control group, the composition of the composition,**is P<0.01; compared with the staphylococcus aureus injection control group,##is P<0.01。
The results show that after 8 hours of tail vein injection of staphylococcus aureus ATCC29213, the mice are found to have symptoms of gradually reduced activity and feeding, erect hair, curly hair, trembling and the like. As shown in figure 1, the death time of the mice is mainly concentrated in 48-72 h after tail vein injection of the bacteria, and all animals in the bacteria injection group die within 72h, which indicates that the staphylococcus aureus ATCC29213 sepsis mouse model is successfully replicated. All animals in the high and low dose groups of the trigeminal ethanol extract survived, and none of the animals died. Compared with a staphylococcus aureus ATCC29213 injection control group, the survival time of mice in both groups of administration groups is obviously prolonged. As can be seen from the table above, the ethanol extract of trifacial bitter has an obvious protective effect on sepsis mice induced by staphylococcus aureus ATCC 29213.
TABLE 2 Effect of ethanol extract of Premna trifida on survival and survival time of 168h septic mice induced by Klebsiella pneumoniae ATCC10031 (S)
Figure BDA0002329893070000061
n＝10)
Figure BDA0002329893070000062
Compared with the blank control group, the composition of the composition,**is P<0.01; compared with a Klebsiella pneumoniae injection control group,##is P<0.01。
The results show that after 8 hours of tail vein injection of Klebsiella pneumoniae ATCC10031, the mice have symptoms of piloerection, gradually reduced activity and feeding, and trembling. As shown in FIG. 2, the death time of the mice is mainly concentrated in 0-24 h after tail vein injection of bacteria, and 9 animals die within 24h, accounting for 90% of the total. All animals in the bacteria injection group died within 72h, indicating that the Klebsiella pneumoniae ATCC10031 sepsis mouse model was successfully replicated. 8 animals in the high-dose trigeminal ethanol extract group (1500mg/kg) survived, the survival rate is 80%, and the survival time of mice in the administration group is obviously prolonged compared with that in the Klebsiella pneumoniae injection control group. As can be seen from the table above, the ethanol extract of Premna trifasciata has obvious protective effect on sepsis mice induced by Klebsiella pneumoniae ATCC 10031.
TABLE 3 Effect of ethanol extract of Evodia trifoliata on 168h survival and survival time of Evodia coli ATCC25922 induced septic mice ((S))
Figure BDA0002329893070000063
n＝10)
Figure BDA0002329893070000064
Figure BDA0002329893070000071
Compared with the blank control group, the composition of the composition,**is P<0.01; compared with the control group injected with the escherichia coli,##is P<0.01。
The results show that after 8 hours of tail vein injection of Escherichia coli ATCC25922, the mice are found to have symptoms of hair erection, gradually reduced activity and reduced food intake, and the like. As shown in FIG. 3, the death time of the mice is mainly concentrated in 0-24 h after tail vein injection of bacteria, and 8 animals die within 24h, accounting for 80% of the total. All animals in the bacteria injection group died within 72h, indicating that the Escherichia coli ATCC25922 sepsis mouse model was successfully replicated. All animals in the high-dose trigeminal ethanol extract group (1500mg/kg) survived, the survival rate is 100%, and the survival time of mice in the administration group is obviously prolonged compared with that in the escherichia coli injection control group. As can be seen from the above table, the ethanol extract of Evodia elata has an obvious protective effect on sepsis mice induced by Escherichia coli ATCC 25922.
TABLE 4 Effect of ethanol extract of trifacial bitter on the survival and survival time of 168h septic mice induced by Pseudomonas aeruginosa ATCC27853: (
Figure BDA0002329893070000072
n＝10)
Figure BDA0002329893070000073
Compared with the blank control group, the composition of the composition,**is P<0.01; compared with the pseudomonas aeruginosa injection control group,##is P<0.01。
The result shows that the mice are found to have symptoms of hair erection, gradual reduction of activity and food intake and the like after 8 hours of tail vein injection of the pseudomonas aeruginosa ATCC 27853. As shown in FIG. 4, the death time of the mice was mainly concentrated in 0-24 h after tail vein injection of the bacteria, and all animals were within 24 h. All animals in the bacteria injection group die within 72h, which indicates that the mouse model of the sepsis caused by the pseudomonas aeruginosa ATCC27853 is successfully replicated. All animals in the low-dose group (750mg/kg) of the evodia lepta ethanol extract die, only 1 animal in the high-dose group (1500mg/kg) of the evodia lepta ethanol extract survives, and compared with the control group injected with the large escherichia coli, the survival time of mice in the two groups of administration groups is not obviously prolonged. As can be seen from the above table, the ethanol extract of evodia lepta has no obvious protective effect on sepsis mice induced by pseudomonas aeruginosa ATCC 27853.
2.2 experiment of therapeutic action of Evodia extract on LPS-induced mouse sepsis
(1) The administration dosage of the medicine of the trigeminal bitter extract (prepared from 1) is as follows: 750mg/kg of ethanol extract of the trifoliate-bitter.
(2) ICR mice (male and female halves), 6-7 weeks old, 20.0 + -2.0 g. The animal feed is fed in an SPF grade barrier system of animals, placed in a dry and clean plastic cage, fed with standard experimental animal feed, and fed with free water.
(3) Experimental methods animals were randomly grouped by weight, 10 animals per group. Experiments are provided with a blank control group, a pathogenic bacterium control group, a low-dose group of the thin evodia lepta ethanol extract and a high-dose group of the thin evodia lepta ethanol extract. LPS (40mg/kg) was injected into the tail vein, and an equal volume of physiological saline was injected into the vehicle control group. Immediately after LPS injection, the trifacial bitter ethanol extract is administrated once by intragastric administration, the intragastric administration dosage of the low-dose group of the trifacial bitter ethanol extract is 750mg/kg, and the intragastric administration dosage of the high-dose group of the trifacial bitter ethanol extract is 1500 mg/kg. The blank control group and the LPS-injected group were simultaneously administered with distilled water at 0.2ml/20 g.
(4) The status and survival of the mice were recorded every 8 hours of observation and the survival rate of the mice was calculated for 168 h.
(5) And the results obtained
TABLE 5 Effect of ethanol extract of Evodia trifoliata on survival and survival time of LPS-induced septic mice 168h ((S))
Figure BDA0002329893070000081
n＝10)
Figure BDA0002329893070000082
P as compared with blank control group<0.01; compared with LPS injection control group, # is P<0.05，##Is P<0.01。
The results show that after 8 hours of LPS tail vein injection, the mice are found to have the symptoms of hair erection, gradually reduced activity and food intake, diarrhea and the like. As shown in FIG. 5, the death time of the mice is mainly concentrated in 0h-24h after tail vein injection of bacteria, and all animals die within 48h, indicating that the LPS sepsis mouse model is successfully replicated. 2 animals survived in the low-dose group (750mg/kg) of the ethanol extract of the trigeminal bitter, and 8 animals survived in the high-dose group (1500mg/kg) of the ethanol extract of the trigeminal bitter, and the survival time of the animals in the two groups is obviously prolonged compared with that of the LPS injection control group. As can be seen from the table above, the ethanol extract of trifurcate bitter has obvious protective effect on LPS-induced sepsis mice at 750-1500 mg/kg.
2.3 Effect of thin Evodia extract on LPS-induced mouse sepsis serum-induced inflammatory cytokine
(1) The administration concentration of the medicine of the thin evodia leaf extract (prepared from 1) is as follows: 750mg/kg of ethanol extract of the trifoliate-bitter.
(2) ICR mice (male and female halves), 6-7 weeks old, 20.0 + -2.0 g. Feeding in SPF barrier system, placing in dry and clean plastic cage, feeding with standard experimental animal feed, and feeding with free water.
(3) The experimental method comprises the following steps of randomly grouping animals according to body weight, and setting a blank control group, a pathogenic bacterium control group, a trigeminal bitter ethanol extract low-dose group and a trigeminal bitter ethanol extract high-dose group in an experiment, injecting LPS (40mg/kg) into tail veins, injecting isovolumetric normal saline into a solvent control group, immediately performing intragastric administration on the trigeminal bitter ethanol extract once after the LPS injection, performing intragastric administration on the trigeminal bitter ethanol extract low-dose group at 750mg/kg, performing intragastric administration on the trigeminal bitter ethanol extract high-dose group at 1500 mg/kg. in the blank control group and the LPS injection group at 0.2ml/20g, simultaneously administering distilled water for 8 hours after the LPS injection, removing venous blood by an eyeball method, centrifuging for 10min at 4 ℃ and 3000 rpm, collecting serum, and using mouse TNF- α and IL-6 detection kits and referring to kit instructions to detect the expression levels of TNF- α and IL-6 in the serum.
(5) And the results obtained
TABLE 6 Effect of ethanol extract of Evodia trifoliata on the expression levels of TNF- α and IL-6 in LPS-induced serum of septic mice ((
Figure BDA0002329893070000101
n＝10)
Figure BDA0002329893070000102
P <0.01 compared to placebo; compared with the LPS injection control group, # # is P < 0.01.
The results show that after 8 hours of LPS tail vein injection, as shown in figure 6 and figure 7, compared with a blank control group, the serum TNF- α and IL-6 levels of mice in the LPS injection group are obviously increased, compared with the LPS injection control group, the high dose (1500mg/kg) of the ethanol extract of trigeminal amaryllin obviously reduces the serum TNF- α and IL-6 levels of sepsis mice, and the table shows that the ethanol extract of trigeminal amaryllin has obvious down-regulation effect on the expression levels of inflammatory cytokines TNF- α and IL-6 in the serum of sepsis mice induced by LPS.
3 cell assay
3.1 Effect of thin Evodia extract on RAW264.7 cell proliferation
(1) The administration concentration of the medicine of the thin evodia leaf extract (prepared from 1) is as follows: the ethanol extract of the evodia lepta ranges from 50 to 800 mu g/ml.
(2) Mouse leukemia RAW264.7 cells, purchased from american specialty storage center (ATCC).
(3) The cells were collected in log phase, the cell suspension concentration was adjusted to 1X 104/ml and 100. mu.l of cell suspension was added per well. 5% CO2, incubated at 37 ℃ for 24 h. Adding ethanol extracts of thin leaf of evodia rutaecarpa with different concentrations to make the final concentration in the wells respectively 50, 100, 200, 400 and 800 μ g/ml, setting up a solvent control well with 100 μ l cell suspension per well and 3 multiple wells per concentration, and incubating at 37 deg.C for 24 hr. Mu.l of MTT solution (5mg/ml) was added to each well and incubation was continued for 4 h. The culture was terminated and the culture medium in the wells was carefully aspirated. Then, 150. mu.l of dimethyl sulfoxide was added to each well to dissolve the crystals sufficiently. The absorbance (OD) of each well was measured at 490nm using a microplate reader. Cell proliferation rate (OD)Treatment of/ODSolvent×100％。
(4) And the results obtained
TABLE 7 Effect of ethanol extract of Evodia trifoliata on RAW264.7 cell proliferation Rate ((S))
Figure BDA0002329893070000111
n＝3)
Figure BDA0002329893070000112
Compared with the solvent control group,*is P<0.05。
The results show that, as shown in fig. 8, the ethanol extract of trifurcate has obvious inhibition effect on RAW264.7 cell proliferation at 800 μ g/ml compared with the vehicle group, but has no obvious effect on cell proliferation at the concentration of 50-400 μ g/ml as the concentration of the ethanol extract of trifurcate is reduced.
3.2 Effect of Amaranthus trifoliatus extract on TNF- α and IL-6 in supernatant of RAW264.7 cells stimulated by LPS
(1) The administration concentration of the medicine of the thin evodia leaf extract (prepared from 1) is as follows: the ethanol extract of the trifoliate bitter is 50, 100 and 200 mu g/ml.
(2) Mouse leukemia RAW264.7 cells, purchased from ATCC.
(3) Collecting cells in logarithmic phase, adjusting cell suspension concentration to 1 × 106Perml, seeded in 90mm cell culture dishes, 10ml of cell suspension, 5% CO per dish2And incubating at 37 ℃, when the confluence degree of cells in a culture dish reaches about 90%, simultaneously adding LPS and different concentrations of the evodia lepta ethanol extracts to ensure that the final concentration of the LPS in the dish reaches 200ng/ml, the final concentrations of the evodia lepta ethanol extracts reach 50, 100 and 200 mu g/ml respectively, simultaneously setting a solvent control dish, arranging 10ml of cell suspension in each dish, setting 3 parallel control dishes, incubating at 37 ℃, after incubating for 8 hours, respectively taking 0.5ml of cell supernatant, using a mouse TNF- α and IL-6 detection kit, referring to the kit specification for operation, and adopting an enzyme labeling instrument to measure the light absorption value (OD) of each hole at 490nm so as to detect the expression levels of TNF- α and IL-6 in the cell supernatant.
(4) And the results obtained
TABLE 8 Effect of ethanol extract of Evodia trifoliata on the expression levels of TNF- α and IL-6 in the supernatant of LPS-stimulated RAW264.7 cells ((
Figure BDA0002329893070000121
n＝3)
Figure BDA0002329893070000122
Compared with the solvent control group,**is P<0.01; compared with the control group stimulated by LPS,#is P<0.05。
The results show that after 8h of LPS stimulation, as shown in figure 9 and figure 10, compared with the vehicle control group, the expressions of TNF- α and IL-6 in the cell supernatant are obviously increased after the LPS stimulation, the expression levels of TNF- α and IL-6 in the cell supernatant of RAW264.7 stimulated by LPS are obviously reduced when the ethanol extract of trigeminal bitter is in high concentration (200 mug/ml), and ELISA tests further prove that the ethanol extract of trigeminal bitter has obvious inhibition effect on cytokines generated by LPS-induced inflammation.
In summary, the following steps: the experimental result of the invention shows that different pathogenic bacteria are injected through tail vein, and the injection dosage is the minimum dosage (100% MLD) of 100% animal death. Wherein, the ratio of staphylococcus aureus ATCC29213 to staphylococcus aureus ATCC29213 is 3.3 multiplied by 1010CFU/ml, Klebsiella pneumoniae ATCC10031: 2.0X 1010CFU/ml, Escherichia coli ATCC25922: 1.5X 1010CFU/ml, Pseudomonas aeruginosa ATCC27853: 1.6X 1010CFU/ml) or 40mg/kg LPS to establish a mouse sepsis model, in addition to Pseudomonas aeruginosa ATCC27853, the trigeminal ethanol extract can obviously improve the survival rate and survival time of bacteria or LPS-induced sepsis mice 168h, in addition, ELISA is adopted to detect the expression levels of inflammatory cytokines TNF- α and IL-6 in the serum of LPS-induced sepsis mice, and the trigeminal ethanol extract can obviously reduce the expression levels of TNF- α and IL-6 in the serum of LPS-induced sepsis mice.
Therefore, the invention provides the application of the ethanol extract of the trifacial bitter in preparing the medicine for treating the sepsis, and provides a new idea for the research and development of the medicine for treating the sepsis. The trifoliate bitter resources are abundant and easy to obtain, and can drive the full development and utilization of medicinal material resources, thereby producing beneficial effects.
Drawings
FIG. 1 is a schematic diagram showing the effect of ethanol extract of evodia lepta on the survival rate of Staphylococcus aureus ATCC29213 sepsis-causing mouse 168h in the experiment of the present invention to prove 2.1;
FIG. 2 is a schematic diagram showing the effect of ethanol extract of Premna trifurcata in 2.1 on the survival rate of Klebsiella pneumoniae ATCC10031 sepsis-induced mouse 168h in the experiment of the invention;
FIG. 3 is a schematic diagram showing that the experiment of the present invention proves the effect of ethanol extract of thin evodia on the survival rate of 168h mice with sepsis caused by Escherichia coli ATCC25922 in 2.1;
FIG. 4 is a schematic diagram showing that the experiment of the invention proves that the influence of the ethanol extract of evodia lepta in 2.1 on the survival rate of a mouse with pseudomonas aeruginosa ATCC27853 sepsis for 168 h;
FIG. 5 is a schematic diagram showing that the experiment of the present invention proves the effect of the ethanol extract of evodia lepta in 2.2 on the survival rate of LPS sepsis-induced mouse 168 h;
FIG. 6 is a schematic diagram showing the effect of ethanol extract of evodia lepta in 2.3 on the expression level of TNF- α in serum of mice with sepsis induced by LPS according to the experiment of the present invention;
FIG. 7 is a schematic diagram showing the effect of ethanol extract of evodia lepta in 2.3 on the expression level of IL-6 in serum of mice with sepsis induced by LPS according to the experimental demonstration of the present invention;
FIG. 8 is a schematic diagram showing the effect of the ethanol extract of evodia lepta in 3.1 on the proliferation rate of RAW264.7 cells according to the experimental demonstration of the present invention;
FIG. 9 is a graph showing the effect of ethanol extract of evodia lepta in 3.2 on the expression level of TNF- α in the supernatant of LPS-stimulated RAW264.7 cells;
FIG. 10 is a graph showing the effect of ethanol extract of Evodia trifoliata in 3.2 on the expression level of IL-6 in the supernatant of RAW264.7 cells stimulated by LPS.
Detailed Description
The present invention is further illustrated by the following examples, which are not to be construed as limiting the invention.
Example 1: pulverizing and sieving thin Evodia leaf, adding 70% ethanol, cold soaking for 3 times, filtering, concentrating the filtrate at 60 deg.C under reduced pressure for 1 time, oven drying at 60 deg.C in water bath, and drying in 60 deg.C reduced pressure oven to obtain thin Evodia leaf ethanol extract; the weight ratio of the evodia lepta to the ethanol is 1:10, and the filtrate is decompressed and concentrated to 1/3 of the original volume. Adding appropriate amount of conventional adjuvants into the ethanol extract of evodia lepta, mixing, granulating, drying, tabletting, and making into tablet. Each tablet contains 500mg of ethanol extract of evodia lepta.
The usage and dosage are as follows: the preparation is taken 4 tablets each time for 3-4 times a day for treating sepsis.
Example 2: pulverizing and sieving thin Evodia leaf, adding 80% ethanol, cold soaking for 4 times, filtering, concentrating the filtrate at 70 deg.C under reduced pressure for 2 times, oven drying at 80 deg.C in water bath, and drying in 70 deg.C reduced pressure oven to obtain thin Evodia leaf ethanol extract; the weight ratio of the evodia lepta to the ethanol is 1:7, and the filtrate is decompressed and concentrated to 1/4 of the original volume. Adding appropriate amount of conventional adjuvants into the ethanol extract of evodia lepta, mixing, oven drying, sterilizing, making into hard capsule, packaging, and making into capsule. Each tablet contains 500mg of ethanol extract of evodia lepta.
The usage and dosage are as follows: the preparation is taken 3-4 times a day, 4 pills each time, and can be used for treating sepsis.
Example 3: pulverizing and sieving thin Evodia leaf, adding 90% ethanol, cold soaking for 5 times, filtering, concentrating the filtrate at 80 deg.C under reduced pressure for 2 times, oven drying at 85 deg.C in water bath, and drying in 80 deg.C reduced pressure oven to obtain thin Evodia leaf ethanol extract; the weight ratio of the evodia lepta to the ethanol is 1:8, and the filtrate is decompressed and concentrated to 1/5 of the original volume. Adding appropriate amount of common adjuvants into the ethanol extract of evodia lepta, granulating, drying, grading, and making into granule. The granule is 5 g/bag, and the content of the ethanol extract of Foliumet ramulus evodiae in the granule is 100 mg/g.
The usage and dosage are as follows: the preparation is taken 3-4 times a day, 4 bags each time, and can be used for treating sepsis.
Example 4: pulverizing and sieving thin Evodia leaf, adding 70% ethanol, cold soaking for 4 times, filtering, concentrating the filtrate at 70 deg.C under reduced pressure for 2 times, oven drying at 80 deg.C in water bath, and drying in 70 deg.C reduced pressure oven to obtain thin Evodia leaf ethanol extract; the weight ratio of the evodia lepta to the ethanol is 1:10, and the filtrate is decompressed and concentrated to 1/4 of the original volume. Adding appropriate amount of solubilizer into the ethanol extract of evodia lepta, grinding, diluting with small amount of water, mixing, adding correctant and antiseptic, mixing, adding water to desired amount, filtering, mixing, packaging, sterilizing, and making into oral liquid. The content of the ethanol extract of the trifacial bitter in the oral liquid is 60 mg/ml.
The usage and dosage are as follows: the preparation is taken 3 times a day, 20-40 ml each time, and is used for treating sepsis.
Example 5: pulverizing and sieving thin Evodia leaf, adding 90% ethanol, cold soaking for 5 times, filtering, concentrating the filtrate at 80 deg.C under reduced pressure for 2 times, oven drying at 85 deg.C in water bath, and drying in 85 deg.C reduced pressure oven to obtain thin Evodia leaf ethanol extract; the weight ratio of the evodia lepta to the ethanol is 1:10, and the filtrate is decompressed and concentrated to 1/5 of the original volume. Adding appropriate amount of common adjuvants into the ethanol extract of evodia lepta, making into pill, drying, and making into pill. Each pill contains 500mg of ethanol extract of Foliumet ramulus evodiae.
The usage and dosage are as follows: the preparation is taken 3-4 times a day, 4 pills each time, and can be used for treating sepsis.
Example 6. Pulverizing Foliumet ramulus evodiae, adding appropriate amount of conventional adjuvants, mixing, granulating, drying, tabletting, and making into tablet. Each tablet contains 1500mg of Foliumet ramulus evodiae.
The usage and dosage are as follows: the preparation is taken 3-4 times a day, 4 tablets each time, and is used for treating sepsis.
Example 7: pulverizing folium evodiae, adding appropriate amount of conventional adjuvants, mixing, oven drying, sterilizing, making into hard capsule, and packaging. Each tablet contains 1500mg of Foliumet ramulus evodiae.
The usage and dosage are as follows: the preparation is taken 3-4 times a day, 4 pills each time, and is used for treating sepsis.
Example 8: pulverizing Foliumet ramulus evodiae, adding appropriate amount of common adjuvants, granulating, drying, grading, and making into granule. The granule is 5 g/bag, and the content of Foliumet ramulus evodiae in the granule is 300 mg/g.
The usage and dosage are as follows: the preparation is taken 3-4 times a day, 4 bags each time, and is used for treating sepsis.
Example 9: pulverizing Foliumet ramulus evodiae, adding appropriate amount of solubilizer, grinding, diluting with small amount of water, mixing, adding correctant and antiseptic, mixing, adding water to desired amount, filtering, mixing, packaging, sterilizing, and making into oral liquid. The content of Foliumet ramulus evodiae in the oral liquid is 180 mg/ml.
The usage and dosage are as follows: the preparation is taken 3 times a day, 20-40 ml each time, and is used for treating sepsis.
Example 10: pulverizing folium evodiae, adding appropriate amount of common adjuvants, making into pill, drying, and making into pill. Each pill contains 1500mg of Foliumet ramulus evodiae.
The usage and dosage are as follows: the preparation is taken 3-4 times a day, 4 pills each time, and is used for treating sepsis.

Claims (10)

1. Application of evodia lepta in preparing medicine for treating sepsis is provided.
2. Application of Foliumet extract in preparing medicine for treating sepsis is provided.
3. The use of an ethanol extract of trifacial bitter as claimed in claim 2 in the manufacture of a medicament for the treatment of sepsis, wherein: the thin evodia leaf extract is thin evodia leaf extract.
4. The use of an ethanol extract of trifacial bitter as claimed in claim 3 in the manufacture of a medicament for the treatment of sepsis, wherein: the ethanol extract of evodia lepta is prepared by pulverizing evodia lepta leaf, sieving, adding 70-90% ethanol, cold soaking for 3-5 times, filtering, concentrating the filtrate at 60-80 deg.C under reduced pressure, and drying at 60-85 deg.C.
5. The use of an ethanol extract of trifacial bitter as claimed in claim 4 in the manufacture of a medicament for the treatment of sepsis, wherein: the ethanol extract of evodia lepta is prepared by pulverizing evodia lepta leaf, sieving, adding 80% ethanol, cold soaking for 4 times, filtering, concentrating the filtrate at 70 deg.C under reduced pressure by distillation for 1-2 times, oven drying at 80 deg.C in water bath, and drying in 70 deg.C reduced pressure oven to obtain evodia lepta ethanol extract.
6. The use of an ethanol extract of trifacial bitter as claimed in claim 4 in the manufacture of a medicament for the treatment of sepsis, wherein: the weight ratio of the evodia lepta to the ethanol is 1:5-1:10, and the filtrate is decompressed and concentrated to 1/3-1/5 of the original volume.
7. A medicament for treating sepsis, which is characterized in that: the raw materials of the medicine comprise the trifoliate bitter or a trifoliate bitter extract.
8. A medicament for treating sepsis, which is characterized in that: the medicine is mainly prepared from the extract of the trifacial bitter or the trifacial bitter.
9. A method for the preparation of a medicament for the treatment of sepsis according to claim 7 or 8, wherein: taking the extract of the trifoliate bitter or the trifoliate bitter, combining the extract with auxiliary materials acceptable in the medicine, processing the mixture according to a conventional method, and preparing the corresponding medicine.
10. The method for preparing a medicament for treating sepsis according to claim 9, wherein: the medicine is tablet, capsule, granule, oral liquid or pill.
CN201911332041.XA 2019-12-21 2019-12-21 Application of ethanol extract of evodia lepta in preparation of medicine for treating sepsis Withdrawn CN111000919A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111072683A (en) * 2020-01-02 2020-04-28 华润三九医药股份有限公司 Coumarin dimer compound, pharmaceutical composition, preparation method and application thereof

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111072683A (en) * 2020-01-02 2020-04-28 华润三九医药股份有限公司 Coumarin dimer compound, pharmaceutical composition, preparation method and application thereof

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