CN112546153A

CN112546153A - Acinetobacter baumannii-resistant composition and preparation method and application thereof

Info

Publication number: CN112546153A
Application number: CN202011439582.5A
Authority: CN
Inventors: 李倩婷; 姜志辉; 苏澎; 李娜
Original assignee: Southern Theater Command General Hospital of PLA
Current assignee: Southern Theater Command General Hospital of PLA
Priority date: 2020-12-11
Filing date: 2020-12-11
Publication date: 2021-03-26

Abstract

The invention relates to the technical field of biology, in particular to an anti-acinetobacter baumannii composition and a preparation method and application thereof. The anti-acinetobacter baumannii composition comprises a mixture of the iphigenia indica and the moutan bark, or extracts of the iphigenia indica and the moutan bark, wherein the extracts of the iphigenia indica and the moutan bark are water extracts or alcohol extracts. The invention also provides an application of the acinetobacter baumannii-resistant composition, which is used for resisting acinetobacter baumannii or preparing acinetobacter baumannii-resistant medicines. According to the application, the treatment of the nematodes infected with the acinetobacter baumannii proves that the extracts of the pleione bulbocodioides and the moutan bark have a good bactericidal effect on the acinetobacter baumannii, provide a good early-stage research foundation for researching and developing novel anti-acinetobacter baumannii medicines, and bring about eosin for solving the difficult problem of clinically treating, preventing and controlling the acinetobacter baumannii.

Description

Acinetobacter baumannii-resistant composition and preparation method and application thereof

Technical Field

The invention relates to the technical field of biology, in particular to an anti-acinetobacter baumannii composition and a preparation method and application thereof.

Background

Acinetobacter baumannii is the most common non-fermentation gram-negative bacterium in acinetobacter, belongs to conditional pathogenic bacteria, and is listed as a super bacterium needing the most advanced solution by the world health organization. The bacterium can survive for a long time in a hospital environment, the clinical department distribution is mainly the most ICU, and the respiratory medicine is the second one. Elderly patients, patients with weak body resistance and critical illness, as well as patients using various invasive procedures and long-term treatment with broad-spectrum antibiotics, are susceptible subjects. Acinetobacter baumannii has the capability of rapid acquisition and transmission, and the Acinetobacter baumannii has a worldwide epidemic trend and becomes one of the most important pathogenic bacteria for nosocomial infection in China. According to the data of the CHINET bacterial monitoring network of 2018, the number of acinetobacter baumannii in 35 education hospitals in China accounts for about 14.7% of that of gram-negative bacteria clinically isolated, and is second only to escherichia coli and klebsiella pneumoniae, so that a novel anti-acinetobacter baumannii medicine is urgently required to be developed.

Many scholars have abundant experience in treating infectious diseases by using traditional Chinese medicines for clearing heat and removing toxicity. As early as in Han Dynasty, Shang Han Lun, many effective prescriptions were summarized, such as Xie Xin Tang, Ge Gen Qin Lian Tang and Bai Tou Weng Tang. At present, a plurality of scholars think that the Shanghai treatise is a special treatise on infectious diseases as the actual classic treatise, and the occurrence of the writings of the Qing Dynasty, the pestilence treatise, the Wen Bing tiao Bian and the like further enriches the treatment experience of the infectious diseases and provides valuable resources for the deep research on the anti-infection treatment of the traditional Chinese medicine. At present, scholars at home and abroad do a lot of work on the research on the antibacterial effect of Chinese herbal medicines, and experiments and clinical research all achieve certain results. However, most of these studies are in vitro antibacterial activity studies, and there is no effective elimination method for compounds with high toxicity, poor pharmacokinetic properties and poor in vitro and in vivo correlation, so that reliable research results have not been obtained.

Disclosure of Invention

The invention aims to provide an anti-acinetobacter baumannii composition which has a good bactericidal effect on acinetobacter baumannii, and also provides a preparation method and application thereof.

The anti-acinetobacter baumannii composition comprises a mixture of the Indian iphigenia bulb and the tree peony bark or extracts of the Indian iphigenia bulb and the tree peony bark.

Preferably, the extracts of the Indian iphigenia bulb and the tree peony bark are water extracts or alcohol extracts; the alcohol extract is more preferable, the paeonol in the tree peony bark is a fat-soluble compound, the paeonol is more easily extracted by the alcohol extract, and the paeonol in the tree peony bark has the in-vivo antibacterial effect, so the effect of the alcohol extract is better than that of the water extract; more preferably 50% by weight.

The application of the acinetobacter baumannii-resistant composition is used for resisting acinetobacter baumannii or preparing acinetobacter baumannii-resistant medicines.

Preferably, the anti-acinetobacter baumannii drug is an in vivo anti-acinetobacter baumannii drug.

The preparation method of the acinetobacter baumannii-resistant composition comprises the following steps:

(1) mixing the mixture of Pseudobulbus Cremastrae Seu pleiones and cortex moutan with solvent, heating and refluxing, separating solid and liquid,

(2) mixing the solid obtained in the step (1) with a solvent, heating and refluxing, carrying out solid-liquid separation,

(3) and (3) mixing the liquid obtained in the step (1) and the liquid obtained in the step (2), and removing the solvent to obtain the extracts of the Indian iphigenia bulb and the tree peony bark.

Wherein: the weight ratio of the edible tulip to the moutan bark in the mixture in the step (1) is 0.2:1-1: 2; more preferably 1:1, and this range of ratios provides better efficacy than other ratios.

The mass ratio of the mixture to the solvent in the step (1) is 1: 5-20; the mass ratio of the solid to the solvent in the step (2) is 1: 5-20.

The solvent in the step (1) and the step (2) is water or alcohol; more preferably, the alcohol is ethanol with the mass fraction of 50% or 95%.

The solvent is water, the heating reflux temperature is 90-100 ℃, and the heating reflux time is 1-2 hours.

The solvent is ethanol, the heating reflux temperature is 80-90 ℃, and the heating reflux time is 1-2 hours.

The experimental model adopted in the invention is a bacterial-nematode infection model, is an in-vivo model, and the pharmacodynamic data obtained on the model is in-vivo pharmacodynamic data.

In summary, the invention has the following advantages:

(1) the invention provides an anti-acinetobacter baumannii composition which has a very good bactericidal effect on acinetobacter baumannii.

(2) The invention provides an application of a iphigenia indica combined cortex moutan extract in resisting acinetobacter baumannii, and comprises treatment of the iphigenia indica combined cortex moutan extract in vivo resisting acinetobacter baumannii. The edible tulip is sweet and slightly pungent in taste and cool in nature, and has the effects of clearing heat and removing toxicity, eliminating carbuncles and dissipating stagnation and the like; cortex moutan is bitter and pungent in flavor and slightly cold in property, and has effects of clearing heat and cooling blood, promoting blood circulation and removing blood stasis, and relieving deficiency heat. The two medicines of the Indian iphigenia bulb and the tree peony bark are compatible and have synergistic action, one is in favor of interior walking and the other is in favor of exterior walking, and the synergistic antibacterial action is played comprehensively.

(3) The preparation method of the edible tulip combined cortex moutan extract disclosed by the invention has the advantages of abundant and easily-obtained raw material resources, low price and simple and easily-operated preparation process.

(4) According to the invention, the extract of the edible tulip combined with the tree peony bark is used for carrying out antibacterial treatment on the nematodes infected with the acinetobacter baumannii, so that the extract of the edible tulip combined with the tree peony bark has a good bactericidal effect on the acinetobacter baumannii, a good early-stage research basis is provided for the research and development of novel medicines for resisting the acinetobacter baumannii, and the eosin is brought for solving the difficult dilemma of clinically treating and preventing and controlling the acinetobacter baumannii.

Detailed Description

It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.

It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments according to the present application. As used herein, the singular forms also include the plural forms unless the context clearly dictates otherwise, and further, it is understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of the stated features, steps, operations, devices, components, and/or combinations thereof.

The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments, and it should be understood that the described embodiments are some, but not all, embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1

An anti-Acinetobacter baumannii composition comprises water extract of Pseudobulbus Cremastrae Seu pleiones and cortex moutan.

The preparation method comprises the following steps:

the first step is as follows: weighing 2g of edible tulip and 2g of moutan bark, placing the mixture in a 100mL round-bottom flask, adding 48g of water according to the mass ratio of 1:12, heating and refluxing the mixture at 100 ℃ for 1.5 hours, filtering the mixture after 1.5 hours, and separating the solid from the liquid.

The second step is that: adding water into the solid obtained in the first step according to the mass ratio of 1:8, heating and refluxing for 1.5 hours at the temperature of 100 ℃, filtering after 1.5 hours, and separating the solid from the liquid.

The third step: and fully mixing the liquid obtained in the first step and the liquid obtained in the second step, and evaporating all solvents under reduced pressure to obtain the medicinal extract. Placing the medicinal extract in a 1.5mL centrifuge tube, and refrigerating and placing in a refrigerator at-20 deg.C to obtain Pseudobulbus Cremastrae Seu pleiones combined cortex moutan water extract.

Example 2

An anti-Acinetobacter baumannii composition comprises alcohol extracts of Pseudobulbus Cremastrae Seu pleiones and cortex moutan.

The preparation method comprises the following steps:

the first step is as follows: weighing 2g of edible tulip and 2g of moutan bark, placing the mixture in a 100mL round-bottom flask, adding 48g of 50% ethanol according to the mass ratio of 1:12, heating and refluxing the mixture at 90 ℃ for 1.5 hours, filtering the mixture after 1.5 hours, and separating the solid from the liquid.

The second step is that: adding water into the solid in the first step according to the mass ratio of 1:8, heating and refluxing for 1.5 hours at the temperature of 90 ℃, filtering after 1.5 hours, and separating the solid from the liquid.

The third step: and fully mixing the liquid obtained in the first step and the liquid obtained in the second step, and evaporating all solvents under reduced pressure to obtain the medicinal extract. Placing the medicinal extract in a 1.5mL centrifuge tube, and refrigerating and placing in a refrigerator at-20 deg.C to obtain 50% ethanol extract of Pseudobulbus Cremastrae Seu pleiones and cortex moutan Radicis.

Example 3

The preparation method comprises the following steps:

the first step is as follows: weighing 2g of edible tulip and 2g of moutan bark, placing the mixture in a 100mL round-bottom flask, adding 48g of 95% ethanol according to the mass ratio of 1:12, heating and refluxing the mixture at 85 ℃ for 1.5 hours, filtering the mixture after 1.5 hours, and separating the solid from the liquid.

The second step is that: adding water into the solid 1 according to the mass ratio of 1:8, heating and refluxing for 1.5 hours at the temperature of 85 ℃, filtering after 1.5 hours, and separating the solid from the liquid.

The third step: and fully mixing the liquid obtained in the first step and the liquid obtained in the second step, and evaporating all solvents under reduced pressure to obtain the medicinal extract. Placing the medicinal extract in a 1.5mL centrifuge tube, and refrigerating and placing in a refrigerator at-20 deg.C to obtain 95% ethanol extract of Pseudobulbus Cremastrae Seu pleiones and cortex moutan.

Extracts of Pseudobulbus Cremastrae Seu pleiones and cortex moutan obtained in examples 1-3 were subjected to antibacterial testing using Acinetobacter baumannii-caenorhabditis elegans infection model:

20% Brain Heart Infusion (BHI) containing 10. mu. mol iron ions was used) The synchronized caenorhabditis elegans is flushed from the nematode growth medium into a 15mL centrifuge tube in the liquid medium, centrifuged for 1 minute at 800rpm, the supernatant is removed, 20% BHI medium containing 10. mu. mol iron ions is added to about 2mL, and the mixture is placed in a thermostat with the temperature of 25 ℃ and the humidity of 85% for standby. A20% BHI medium containing 10. mu. mol iron ions was added to a 3mL turbidimetric tube, and a single colony of Acinetobacter baumannii was picked from a Luria-Bertan (LB) agar plate with a sterile cotton swab, thoroughly mixed in the turbidimetric tube, and the concentration of the bacteria was adjusted to 0.5 M.unit (about 1X 108CFU/mL) using a turbidimeter. Using a pipette gun to suck 250 mu L of liquid with the adjusted bacterium concentration, adding the liquid into a new 5mL centrifuge tube, adding 2250 mu L of 20% BHI liquid culture medium containing 10 mu mol of iron ions, and diluting the bacterium liquid by 10 times to 1X 10⁷CFU/mL. 2mL of the diluted bacterial solution was aspirated by a 1000. mu.L pipette and added to a nematode tube to give a final Acinetobacter baumannii concentration of 5X 10⁶CFU/mL, then placing into a constant temperature and humidity incubator with the temperature of 25 ℃ and the humidity of 85% for culture. After the co-incubation infection is carried out for 6 hours, washing for at least 3 times by using M9 liquid culture medium, sucking 180 mu L of solution and distributing into a 96-well plate, wherein 15-20 caenorhabditis elegans infected by acinetobacter baumannii in each well are suitable, and an acinetobacter baumannii-caenorhabditis elegans infection model is obtained. The fluorescent bacteria are used for infecting the nematodes, and a large amount of bacteria are found to be accumulated in intestinal tracts of the nematodes, so that the acinetobacter baumannii infected by the nematodes in the obtained acinetobacter baumannii-caenorhabditis elegans infection model can be considered as in vivo infection.

The in vivo antibacterial activity of the edible tulip combined cortex moutan extract is detected by using an acinetobacter baumannii-caenorhabditis elegans infection model.

1. Experimental group

Three parallel experiments were performed using different concentrations (1000. mu.g/mL, 100. mu.g/mL, 10. mu.g/mL, 1. mu.g/mL, 0.1. mu.g/mL) of the aqueous extract of the combination of Pseudobulbus Cremastrae Seu pleiones prepared in example 1, the 50% ethanol extract of the combination of Pseudobulbus Cremastrae Seu pleiones prepared in example 2, and the 95% ethanol extract of the combination of Pseudobulbus Cremastrae Seu pleiones prepared in example 3 as the experimental groups.

The preparation method of the iphigenia indica combined moutan bark water extract with different concentrations comprises the following steps: weighing 10mg of extract, placing the extract in a 1.5mL centrifuge tube, adding 100 μ L of water, adding 900 μ L of BHI culture medium, fully mixing and dissolving the extract, sucking 100 μ L of mixed solution, adding 900 μ L of BHI culture medium, and diluting for 5 times to obtain the Pseudobulbus Cremastrae Seu pleiones combined cortex moutan water extractive solutions with concentrations of 1000 μ g/mL, 100 μ g/mL, 10 μ g/mL, 1 μ g/mL and 0.1 μ g/mL respectively.

The preparation method of 50% ethanol extracts of edible tulip and moutan bark in combination with different concentrations comprises the following steps: weighing 10mg of extract, placing the extract in a 1.5mL centrifuge tube, adding 100 μ L of 50% ethanol, adding 900 μ L of BHI culture medium, mixing thoroughly to dissolve the extract, sucking out 100 μ L of mixed solution, adding 900 μ L of BHI culture medium, and diluting for 5 times to obtain 50% ethanol extractive solutions of Pseudobulbus Cremastrae Seu pleiones combined with cortex moutan with concentrations of 1000 μ g/mL, 100 μ g/mL, 10 μ g/mL, 1 μ g/mL, and 0.1 μ g/mL respectively.

The preparation method of the edible tulip bulb and moutan bark 95% ethanol extract with different concentrations comprises the following steps: weighing 10mg of extract, placing the extract in a 1.5mL centrifuge tube, adding 100 μ L of 95% ethanol, adding 900 μ L of BHI culture medium, fully mixing and dissolving the extract, sucking 100 μ L of mixed solution, adding 900 μ L of BHI culture medium, and diluting for 5 times to obtain 95% ethanol extractive solutions of Pseudobulbus Cremastrae Seu pleiones with cortex moutan, wherein the concentrations of the 95% ethanol extractive solutions are 1000 μ g/mL, 100 μ g/mL, 10 μ g/mL, 1 μ g/mL and 0.1 μ g/mL respectively.

2. Positive control group

Polymyxin B at 20. mu.g/mL was used as a positive control.

10mg of polymyxin B is weighed and placed in a 1.5mL centrifuge tube, 1mL of BHI culture medium is added to mix well and dissolve the extract, 100 μ L of the mixture is sucked out and 900 μ L of BHI culture medium is added to dilute for 5 times, and polymyxin B solutions with the concentrations of 1000 μ g/mL, 100 μ g/mL, 10 μ g/mL, 1 μ g/mL and 0.1 μ g/mL are obtained respectively.

3. Negative control group

10% dimethyl sulfoxide was used as a negative control.

10mg of 10% dimethyl sulfoxide was weighed and placed in a 1.5mL centrifuge tube, 1mL of BHI medium was added to mix well and dissolve the extract, 100. mu.L of the mixture was aspirated and added to 900. mu.L of BHI medium to dilute 5 times, and dimethyl sulfoxide solutions with concentrations of 1000. mu.g/mL, 100. mu.g/mL, 10. mu.g/mL, 1. mu.g/mL, and 0.1. mu.g/mL were obtained, respectively.

20 mu L of prepared iphigenia indica combined cortex moutan extract is respectively added into a hole containing caenorhabditis elegans infected with acinetobacter baumannii, the iphigenia indica combined cortex moutan extract is respectively used for intervening with 1000 mu g/mL, 100 mu g/mL, 10 mu g/mL, 1 mu g/mL and 0.1 mu g/mL gradient final concentration extracts, after incubation for 36h in a thermostat at 25 ℃, the treatment effect of the extracts with different concentrations on the caenorhabditis elegans infected with acinetobacter baumannii is judged according to the survival rate of the nematodes, the experimental results are shown in table 1, and table 1 shows the influence condition of the iphigenia indica combined cortex moutan extract with different concentrations on the survival of the acinetobacter baumannii nematodes.

TABLE 1

The result shows that the iphigenia indica combined cortex moutan extract, including water extract and alcohol extract, has a relatively obvious anti-acinetobacter baumannii effect, and the treatment effect is better when the concentration is high than when the concentration is low. The edible tulip and 50% ethanol extract of the moutan bark has better treatment effect compared with a water extract and a 95% ethanol extract no matter under low concentration or high concentration, the survival rate of the infected nematodes can completely survive under the highest concentration, the paeonol in the moutan bark is a fat-soluble compound, the paeonol is more easily extracted by the alcohol solution, and the paeonol in the moutan bark plays a role in vivo antibiosis, so the antibacterial effect of the alcohol extract of the invention is better than that of the water extract.

Comparative example 1

A Pseudobulbus Cremastrae Seu pleiones extract is prepared by the following steps:

the first step is as follows: weighing three parts of 4g of edible tulip, placing the weighed three parts into 3 100mL round-bottom flasks, adding 48g of water, 50% of ethanol by mass fraction and 95% of ethanol by mass fraction according to the mass ratio of 1:12, heating and refluxing the mixture at 100 ℃, 90 ℃ and 85 ℃ for 1.5 hours, filtering the mixture after 1.5 hours, and separating solid from liquid.

The second step is that: adding corresponding water, 50% ethanol and 95% ethanol into the solid obtained in the first step according to the mass ratio of 1:8, heating and refluxing at 100 ℃, 90 ℃ and 85 ℃ for 1.5 hours, filtering after 1.5 hours, and separating the solid from the liquid.

The third step: and (3) fully mixing the liquid obtained in the first step and the liquid obtained in the second step, and evaporating all solvents under reduced pressure to obtain the medicinal extract. Respectively placing the medicinal extracts in 1.5mL centrifuge tubes, and refrigerating and placing in a refrigerator at-20 deg.C to obtain Pseudobulbus Cremastrae Seu pleiones water extract, Pseudobulbus Cremastrae Seu pleiones 50% ethanol extract and Pseudobulbus Cremastrae Seu pleiones 50% ethanol extract.

The in vivo antibacterial activity of the pleione indica extract is detected by utilizing an acinetobacter baumannii-caenorhabditis elegans infection model.

Experiments were performed with different concentrations (1000. mu.g/mL, 100. mu.g/mL, 10. mu.g/mL, 1. mu.g/mL, 0.1. mu.g/mL) of aqueous extract of Pseudobulbus Cremastrae Seu pleiones, 50% ethanol extract of Pseudobulbus Cremastrae Seu pleiones and 95% ethanol extract of Pseudobulbus Cremastrae Seu pleiones, each extract being set up in 3 parallel experiments.

The preparation methods of the edible tulip water extract, the edible tulip 50% ethanol extract and the edible tulip 95% ethanol extract with different concentrations (1000 mug/mL, 100 mug/mL, 10 mug/mL, 0.1 mug/mL) are consistent with the preparation methods of the edible tulip combined cortex moutan water extract, the edible tulip combined cortex moutan 50% ethanol extract and the edible tulip combined cortex moutan 95% ethanol extract with different concentrations in the examples. The experimental results are shown in table 2, and table 2 shows the effect of the Pseudobulbus cremastrae seu pleiones extract on the survival of a nematode infected with Acinetobacter baumannii at different concentrations.

TABLE 2

The results show that the iphigenia indica extract has little influence on the survival rate of the acinetobacter baumannii nematodes, although the iphigenia indica water extract has a slightly better antibacterial effect on the acinetobacter baumannii nematodes at a low concentration than the acinetobacter baumannii nematodes in the example 1 of the invention, the effect of the example 1 of the invention is better than that of a comparative example at a high concentration, and the antibacterial effect of the iphigenia indica combined cortex moutan alcohol extract is far better than that of the iphigenia indica alcohol extract.

Comparative example 2

A cortex moutan extract is prepared by the following steps:

the first step is as follows: weighing three parts of 4g of tree peony bark respectively, placing the tree peony bark in 3 100mL round-bottom flasks, adding 48g of water, 50% of ethanol by mass fraction and 95% of ethanol by mass fraction according to the mass ratio of 1:12 respectively, heating and refluxing the mixture at 100 ℃, 90 ℃ and 85 ℃ respectively for 1.5 hours, filtering the mixture after 1.5 hours, and separating solid from liquid.

The third step: and (3) fully mixing the liquid obtained in the first step and the liquid obtained in the second step, and evaporating all solvents under reduced pressure to obtain the medicinal extract. Respectively placing the medicinal extracts in 1.5mL centrifuge tubes, and refrigerating in a refrigerator at-20 deg.C to obtain cortex moutan water extract, cortex moutan 50% ethanol extract and cortex moutan 50% ethanol extract.

Detecting the in vivo antibacterial activity of cortex moutan extract by using Acinetobacter baumannii-caenorhabditis elegans infection model.

Experiments were performed with different concentrations (1000. mu.g/mL, 100. mu.g/mL, 10. mu.g/mL, 1. mu.g/mL, 0.1. mu.g/mL) of moutan bark water extract, moutan bark 50% ethanol extract and moutan bark 95% ethanol extract, each extract setting 3 sets of parallel experiments.

The preparation methods of the moutan bark water extract, the moutan bark 50% ethanol extract and the moutan bark 95% ethanol extract with different concentrations (1000 mug/mL, 100 mug/mL, 10 mug/mL, 1 mug/mL and 0.1 mug/mL) are consistent with the preparation methods of the iphigenia indica combined moutan bark water extract, the iphigenia indica combined moutan bark 50% ethanol extract and the iphigenia indica combined moutan bark 95% ethanol extract with different concentrations in the embodiment, the experimental results are shown in Table 3, and the Table 3 is a table of the influence of the moutan bark extract on the survival of the acinetobacter baumannii nematode infected with different concentrations.

TABLE 3

The result shows that the cortex moutan water extract has little influence on the survival rate of the acinetobacter baumannii nematode infection and has a common antibacterial effect. Although the antibacterial effect of the 95% alcohol extract of the tree peony bark is slightly better than that of the 95% alcohol extract of the Indian iphigenia bulb and the tree peony bark under the conditions of 10 mu g/mL and 100 mu g/mL, the antibacterial effect of the 95% alcohol extract of the Indian iphigenia bulb and the tree peony bark is better as a whole under the conditions of lower concentration and higher concentration, and more importantly, the antibacterial effect of the 50% alcohol extract and the water extract of the Indian iphigenia bulb and the tree peony bark is obviously better than that of the corresponding 50% alcohol extract and the water extract of the tree peony bark.

Comparative example 3

The preparation of forsythia suspense extract and the detection of its bacteriostatic activity in vivo were carried out according to the method of example 1, using forsythia suspense extract with better anti-acinetobacter baumannii activity in vitro, which has been verified in the art, as a control.

A fructus forsythiae extract is prepared by the following steps:

the first step is as follows: weighing three parts of 2g of fructus forsythiae respectively, placing the fructus forsythiae in 3 100mL round-bottom flasks, adding 48g of water, 50% of ethanol by mass fraction and 95% of ethanol by mass fraction according to the mass ratio of 1:12, heating and refluxing the fructus forsythiae at 100 ℃, 90 ℃ and 85 ℃ respectively for 1.5 hours, filtering the mixture after 1.5 hours, and separating solid from liquid.

The third step: and (3) fully mixing the liquid obtained in the first step and the liquid obtained in the second step, and evaporating all solvents under reduced pressure to obtain the medicinal extract. Respectively placing the medicinal extracts in 1.5mL centrifuge tubes, and refrigerating and placing in a refrigerator at-20 deg.C to obtain fructus forsythiae water extract, fructus forsythiae 50% ethanol extract and fructus forsythiae 50% ethanol extract.

And (3) detecting the antibacterial activity in the forsythia suspense extract in vivo by using an acinetobacter baumannii-caenorhabditis elegans infection model.

Experiments were performed with different concentrations (1000. mu.g/mL, 100. mu.g/mL, 10. mu.g/mL, 1. mu.g/mL, 0.1. mu.g/mL) of aqueous extract of Forsythia suspensa, 50% ethanol extract of Forsythia suspensa, and 95% ethanol extract of Forsythia suspensa, each extract being set up in 3 parallel experiments. The experimental results are shown in table 4, and table 4 shows the effect of forsythia suspense extract on the survival of acinetobacter baumannii nematode infection at different concentrations.

TABLE 4

The results show that the forsythia suspense extract has little influence on the survival rate of the acinetobacter baumannii nematode infection, and has no in-vivo anti-acinetobacter baumannii effect.

Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. An anti-acinetobacter baumannii composition, which comprises a mixture of Indian iphigenia bulb and moutan bark, or an extract of Indian iphigenia bulb and moutan bark.

2. The acinetobacter baumannii composition of claim 1, wherein the extract of iphigenia indica and moutan cortex radicis is an aqueous extract or an alcoholic extract.

3. Use of the Acinetobacter baumannii composition according to claim 1 or 2 for preparing an Acinetobacter baumannii or an anti-Acinetobacter baumannii drug.

4. The use of the Acinetobacter baumannii composition according to claim 3, wherein the Acinetobacter baumannii drug is an in vivo anti-Acinetobacter baumannii drug.

5. A method for preparing an acinetobacter baumannii composition according to claim 1 or 2, comprising the steps of:

6. The method of preparing an acinetobacter baumannii composition according to claim 5, wherein: the weight ratio of the edible tulip bulb to the tree peony bark in the mixture in the step (1) is 0.2:1-1: 2.

7. The method of preparing an acinetobacter baumannii composition according to claim 5, wherein: the mass ratio of the mixture to the solvent in the step (1) is 1: 5-20; the mass ratio of the solid to the solvent in the step (2) is 1: 5-20.

8. The method of preparing an acinetobacter baumannii composition according to claim 5, wherein: the solvent in the step (1) and the step (2) is water or alcohol.

9. The method of preparing an acinetobacter baumannii composition according to claim 8, wherein: the solvent is water, the heating reflux temperature is 90-100 ℃, and the heating reflux time is 1-2 hours.

10. The method of preparing an acinetobacter baumannii composition according to claim 8, wherein: the solvent is ethanol, the heating reflux temperature is 80-90 ℃, and the heating reflux time is 1-2 hours.