阅读说明：本技术 一种抑制铜绿假单胞菌生长的天然组合物及其制备方法和应用 (Natural composition for inhibiting growth of pseudomonas aeruginosa as well as preparation method and application thereof ) 是由 彭红 周刚 王颖思 施庆珊 谢小保 于 2019-11-20 设计创作，主要内容包括：本发明公开了一种抑制铜绿假单胞菌生长的天然组合物及其制备方法和应用。本发明将蜂蜜和苦楝果水煎提取物按照质量比为(1～8):(1～8)的比例混合均匀,得到有效组合物。通过本发明的试验研究表明,蜂蜜和苦楝果水煎提取物对铜绿假单胞菌有优异的抑菌作用,二者复配使用抑制铜绿假单胞菌的生长更高效、更长效,二者复配有协同增效作用。本发明的组合物的原材料天然环保,抑菌效果显著,有益于人体健康,可应用于制备清洗伤口的药物、食品保鲜防腐剂、日化产品抗菌剂等。(The invention discloses a natural composition for inhibiting growth of pseudomonas aeruginosa, a preparation method and application thereof. The effective composition is obtained by uniformly mixing honey and the chinaberry fruit water decoction extract according to the mass ratio of (1-8) to (1-8). Experimental research shows that the honey and the chinaberry fruit water decoction extract have excellent bacteriostatic action on pseudomonas aeruginosa, the combination of the honey and the chinaberry fruit water decoction extract has higher and longer effect on inhibiting the growth of the pseudomonas aeruginosa, and the combination of the honey and the chinaberry fruit water decoction extract has a synergistic effect. The composition has natural and environment-friendly raw materials and remarkable bacteriostatic effect, is beneficial to human health, and can be applied to preparation of medicaments for cleaning wounds, food fresh-keeping preservatives, daily chemical product antibacterial agents and the like.)

1. The antibacterial composition is characterized by comprising honey and a chinaberry extract, wherein the mass ratio of the honey to the chinaberry extract is (1-8) to (1-8).

2. The bacteriostatic composition according to claim 1, wherein the mass ratio of the honey to the chinaberry extract is 1: 1.

3. The bacteriostatic composition according to claim 2, wherein the total mass fraction of the honey and the chinaberry extract in the bacteriostatic composition is 25-33.3%.

4. The bacteriostatic composition according to claim 1, wherein the chinaberry extract is a water-decocted chinaberry fruit extract.

5. The bacteriostatic composition according to claim 4, wherein the water extract of Melia azedarach is prepared by the following steps: oven drying fructus Toosendan, pulverizing, decocting in water, filtering to obtain decoction, concentrating the decoction to 1g per 1mL, and sterilizing to obtain fructus Toosendan water decoction extract.

6. The bacteriostatic composition according to claim 5, wherein the water decoction is obtained by decocting 2 times with water, filtering, and combining the water decoctions for 2 times.

7. The bacteriostatic composition according to any one of claims 1 to 6, wherein the bacterium is Pseudomonas aeruginosa.

8. A method of preparing a bacteriostatic composition according to claim 1, comprising the steps of:

s1: drying and crushing the chinaberry fruits, adding water for decoction, filtering to obtain a water decoction, concentrating the water decoction until each 1mL of the water decoction is equal to 1g of the raw medicinal material, and sterilizing to obtain a chinaberry fruit water decoction extract;

s2: mixing honey and the chinaberry fruit water decoction extract according to the mass ratio of (1-8) to obtain the antibacterial composition.

9. The method as claimed in claim 8, wherein the antibacterial composition is obtained by mixing honey and the water extract of chinaberry fruit in a mass ratio of 1: 1.

10. Use of a bacteriostatic composition according to any one of claims 1 to 6 in the preparation of a medicament for wound cleansing, a food preservative or a daily chemical product antimicrobial.

Technical Field

The invention belongs to the fields of medicine, daily chemicals, animals, plants and microorganisms, and particularly relates to a natural composition for inhibiting growth of pseudomonas aeruginosa, and a preparation method and application thereof.

Background

In recent years, antibiotic resistance is on the rise, new products with remarkable bacteriostatic function are screened from natural animal and plant extracts, strong attention is paid to people, and a therapeutic substitute with bacteriostatic property is paid attention.

A great number of special plants with bacteriostatic ability exist in nature, such as Melia azedarach, Melia toosendan and Neem which are Meliaceae plants, have few parasites, and are high-activity natural bactericides. A great deal of intensive research is carried out on the melia azedarach (mainly the neem) in countries such as America, India and the like, Chinese scholars obtain remarkable results on the research on the aspects of chemical components, pharmacology, sterilization and insecticidal action and the like of the melia azedarach, and the furantriterpenoid compound Toosendanin (Toosendanin) is extracted from barks and fruits of the melia azedarach. The melia azedarach has the effects of dust retention, bacteriostasis, air purification and the like, is easy to decompose in a natural environment, and is safe and environment-friendly. The studies of researchers find that the ethanol extract of the chinaberry tree has good bacteriostatic activity on aspergillus niger and trichoderma viride, and the chinaberry infusion has certain bacteriostatic effect on trichophyton xanthum, trichophyton concentricum, sphaerotheca scholaris, trichophyton rubrum, trichophyton asteroideum and the like.

The use history of honey as an antibacterial drug is long, a large number of folk methods for treating diseases by using honey are kept all over the world to date, and the medicinal value of honey is gradually proved by various research results all over the world in recent years. In 2007, researchers at state university of pennsylvania in the united states published a research result that honey is effective in relieving cough in children, and in 2008, researchers at canada also published a research result that honey has a strong bactericidal effect better than an antibiotic substance, and these results all confirm that honey has an excellent bacteriostatic or bactericidal effect. The honey therapy is a new method for treating wounds in recent years, and researches show that the honey not only has the effects of antibiosis, inflammation diminishing, antioxidation, immunoregulation, autolysis debridement and deodorization, but also has the effect of inhibiting a chronic wound bacterial biofilm. Honey also has the effect of inhibiting bacterial biofilm in chronic wounds. Manuka honey (Manuka honey) is reported in many foreign researches, the Manuka honey has the effects of disinfecting and repairing wounds, and the Manuka honey with the concentration of 10% (w/v) can obviously reduce the biofilm development of two strains of Proteus mirabilis (Proteus mirabilis) and Enterobacter cloacae (Enterobacter cloacae) which are wound separating strains. According to foreign scholars, the honey contains methylglyoxal which is reported to be an antibacterial compound and proved to be a cause of bacterial death in wounds wrapped by biofilms. Foreign scholars study the influence of the combination of honey and antibiotics on the in vitro activity of bacterial biofilms, and find that the index of the combination of vancomycin and honey on staphylococcus aureus biofilms is 0.34, so that the synergistic effect is achieved, and the index of the combination of gentamicin and honey on pseudomonas aeruginosa biofilms is 0.78-0.82, so that the additive effect is achieved.

Pseudomonas aeruginosa (Pseudomonas aeruginosa) is a common pathogenic bacterium. The bacterium is widely distributed in nature, is fond of a humid environment, and exists in various water, soil, air, skin, subcutaneous tissues, bones, ears, eyes, urinary tracts, heart valves, respiratory tracts, intestinal tracts and the like of normal people. The bacterium is one of the main pathogenic bacteria of nosocomial infections, has the characteristics of ring-shaped abscess caused by typical gram-negative bacilli, is usually accompanied by the existence of more virulent bacteria in a focus, and occasionally can cause the infection of tissues exposed to the outside alone. Many infections caused by the bacterium occur in weak or immunocompromised inpatients, are the second most common pathogenic bacteria of intensive care unit infection, patients suffering from metabolic diseases, hematopathy and malignant tumors, and patients after operation or after certain treatments are very easy to infect the bacterium, the infection of wound after operation is often caused, burn wound is also easily infected by copper green pseudomonas, bedsore, abscess, suppurative otitis media and the like can also be caused, and the caused infection focus can cause bacteremia and septicemia and can cause death. There is therefore a need to screen for drugs or methods that inhibit the growth of pseudomonas aeruginosa, thereby reducing the risk of this bacterium.

Disclosure of Invention

The invention aims to overcome the defects in the prior art and provides a natural composition for inhibiting the growth of pseudomonas aeruginosa and a preparation method and application thereof.

The first purpose of the invention is to provide a bacteriostatic composition, which comprises honey and a chinaberry extract, wherein the mass ratio of the honey to the chinaberry extract is (1-8) to (1-8).

The mass ratio of the honey to the chinaberry extract is 1: 1.

The antibacterial composition comprises 25-33.3% of total mass of honey and chinaberry extract.

The chinaberry extract is a water decoction extract of chinaberry fruits.

The honey is natural honey.

The preparation method of the water decoction extract of the melia azedarach fruit comprises the following steps: oven drying fructus Toosendan, pulverizing, decocting in water, filtering to obtain decoction, concentrating the decoction to 1g per 1mL, and sterilizing to obtain fructus Toosendan water decoction extract.

Preferably, the bacterium is Pseudomonas aeruginosa (P.aeruginosa ATCC 9027)

The second purpose of the invention is to provide a preparation method of the antibacterial composition, which comprises the following steps:

s1: drying and crushing the chinaberry fruits, adding water for decoction, filtering to obtain a water decoction, concentrating the water decoction until each 1mL of the water decoction is equal to 1g of the raw medicinal material, and sterilizing to obtain a chinaberry fruit water decoction extract;

s2: mixing honey and the chinaberry fruit water decoction extract according to the mass ratio of (1-8) to obtain the antibacterial composition.

The decoction is prepared by decocting in water for 2 times, filtering, and mixing decoctions for 2 times.

The third purpose of the invention is to provide the application of the antibacterial composition in preparing medicines for cleaning wounds, food fresh-keeping preservatives or daily chemical product antibacterial agents.

The composition obtained by compounding the honey and the chinaberry fruit water decoction extract has a high-efficiency and long-acting antibacterial effect on pseudomonas aeruginosa compared with a single use, and the compound has a synergistic effect.

Experimental research shows that the honey and the chinaberry fruit water decoction extract are compounded and used according to the mass ratio of 1:1, even if the honey and the chinaberry fruit water decoction extract are diluted to 1/8 (mass ratio), the inhibition rates of pseudomonas aeruginosa (P.aeruginosa ATCC 9027) for 5h, 24h, 48h and 5d are respectively 95.15%, 95.76%, 98.25% and 99.99%, and the bacteriostasis effect is excellent.

The raw materials of the composition are derived from animals and plants, are natural and environment-friendly, have remarkable bacteriostatic effect, are beneficial to human health, can be applied to preparation of medicaments for cleaning wounds, food fresh-keeping preservatives or daily chemical product antibacterial agents, and have wide application range and prospect.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and the described embodiments are only a part of the embodiments of the present invention, but not all of the embodiments. The following examples are further illustrative of the present invention and are not intended to be limiting thereof.

Weighing Mel A100 g; weighing 100g of chinaberry fruit in 1000mL of distilled water, soaking for 30min at room temperature, boiling for 30min, decocting, slightly cooling, filtering to obtain a water decoction, adding 500mL of distilled water into the residue, boiling for 30min, filtering to obtain a water decoction, mixing the two water decoctions, concentrating (e.g. concentrating under reduced pressure) the water decoction to 1mL of the water decoction which is equivalent to 1g of the original medicinal material, sterilizing at 121 ℃ for 20min to obtain a chinaberry fruit water decoction extract B, and storing in a refrigerator at 4 ℃ for later use.

When the test strain in the following examples is pseudomonas aeruginosa (p.aeruginosa ATCC 9027), the test method is: the test strains were cultured at 37 ℃ for 1 day using the nutrient agar medium (NA) slant medium in this example. Then washing the bacterial sludge with PBS, diluting to OD₆₀₀And (4) preparing a bacterial suspension working solution as 1.0. A, B are respectively diluted by distilled water, A, B after dilution is combined according to the mass ratio of 1:1 to obtain the A + B composition, and meanwhile, sterile distilled water is used as a blank control. Weighing 9mL of each diluted liquid medicine or blank control into each sterile test tube, placing the sterile test tube in a water bath at 20 +/-1 ℃ for 5min, respectively adding 1mL of bacterial suspension, gently shaking and uniformly mixing, culturing at 37 ℃, performing bacteriostasis test, acting for 5h, 24h, 48h and 5d of test preset time, respectively performing viable bacteria culture counting on the pseudomonas aeruginosa in each test tube, comparing the content of the pseudomonas aeruginosa in the blank control at corresponding time each time, and calculating the bacteriostasis rate.

The formula for calculating the bacteriostasis rate is as follows:

each treatment was done in 3 replicates.

The following abbreviations are used in the specification: ATCC is a shorthand for American type culture Collection, PBS is phosphate buffered saline (phosphate buffer saline), temperature is given in degrees Celsius (C.), min, d day, mL mL mg, g, OD₆₀₀Refers to the absorbance of the solution at a wavelength of 600 nm.