阅读说明：本技术 一种用于预防和/或治疗苯并芘暴露的产品 (Product for preventing and/or treating benzopyrene exposure ) 是由 于雷雷 田丰伟 张凌宇 翟齐啸 陆文伟 崔树茂 王刚 赵建新 张灏 陈卫 于 2020-07-13 设计创作，主要内容包括：本发明公开了一种用于预防和/或治疗苯并芘暴露的产品,属于微生物技术领域以及医药技术领域。本发明提供了一种可预防和/或治疗苯并芘暴露的药品,此药品的有效成分为植物乳杆菌(Lactobacillus plantarum)CCFM8661,此植物乳杆菌CCFM8661能够有效缓解苯并芘暴露,具体体现在：显著降低苯并芘暴露小鼠粪便中苯并芘代谢物3-羟基苯并芘的含量；显著降低苯并芘暴露小鼠血清中谷草转氨酶(AST)和谷丙转氨酶(ALT)的活性；显著降低苯并芘暴露小鼠脑组织中超氧化物歧化酶(SOD)和丙二醛(MDA)的水平；显著改善并芘暴露小鼠的行为学指标；显著提高苯并芘暴露小鼠粪便中短链脂肪酸的含量。(The invention discloses a product for preventing and/or treating benzopyrene exposure, and belongs to the technical field of microorganisms and medicines. The invention provides a medicine capable of preventing and/or treating benzopyrene exposure, the effective component of the medicine is Lactobacillus plantarum (Lactobacillus plantarum) CCFM8661, and the Lactobacillus plantarum CCFM8661 can effectively relieve the benzopyrene exposure, and is specifically represented as follows: the content of benzopyrene metabolite 3-hydroxy benzopyrene in the mouse feces exposed by benzopyrene is remarkably reduced; the activity of glutamic-oxaloacetic transaminase (AST) and glutamic-pyruvic transaminase (ALT) in the serum of a benzopyrene-exposed mouse is obviously reduced; the levels of superoxide dismutase (SOD) and Malondialdehyde (MDA) in the brain tissue of the mouse exposed by benzopyrene are obviously reduced; the behavioral indexes of the pyrene-exposed mice are obviously improved; the content of short-chain fatty acid in the feces of the mice exposed by benzopyrene is obviously improved.)

1. The application of the lactobacillus plantarum in the preparation of the medicine for preventing and/or treating benzopyrene exposure is characterized in that the lactobacillus plantarum is preserved in Guangdong province microorganism strain preservation center with the preservation number of CGMCC No.5494 and the preservation date of 2011, 11 and 29 days.

2. The use according to claim 1, wherein the number of viable Lactobacillus plantarum in the medicament is not less than 1 x 10⁶CFU/mL or 1X 10⁶CFU/g。

3. Use according to claim 1 or 2, wherein the medicament comprises lactobacillus plantarum, a pharmaceutical carrier and/or a pharmaceutical excipient.

4. The use of claim 3, wherein the drug carrier comprises a microcapsule, microsphere, nanoparticle and/or liposome.

5. The use according to claim 3 or 4, wherein the pharmaceutical excipient comprises an excipient and/or an additive.

6. The use of any one of claims 1 to 5, wherein the medicament is in the form of a powder, granules, capsules, tablets, pills or oral liquid.

7. A drug for preventing and/or treating benzopyrene exposure, characterized in that the drug contains lactobacillus plantarum; the lactobacillus plantarum is preserved in the Guangdong province microorganism strain preservation center, the preservation number is CGMCC No.5494, and the preservation date is 2011, 11 and 29 days.

8. The drug for preventing and/or treating benzopyrene exposure according to claim 7, wherein the viable count of Lactobacillus plantarum is not less than 1 x 10⁶CFU/mL or 1X 10⁶CFU/g。

9. The application of the lactobacillus plantarum in removing benzopyrene is characterized in that the lactobacillus plantarum is preserved in Guangdong province microorganism strain preservation center with the preservation number of CGMCC No.5494 and the preservation date of 2011, 11 and 29 days.

10. A benzopyrene scavenger is characterized in that the benzopyrene scavenger contains lactobacillus plantarum; the lactobacillus plantarum is preserved in the Guangdong province microorganism strain preservation center, the preservation number is CGMCC No.5494, and the preservation date is 2011, 11 and 29 days.

Technical Field

The invention relates to a product for preventing and/or treating benzopyrene exposure, belonging to the technical field of microorganisms and medicines.

Background

Benzopyrene (benzopyrene) is a polycyclic aromatic hydrocarbon consisting of 5 benzene rings and has a molecular formula of C₂₀H₁₂The molecular weight is 252.30, the crystal is colorless to light yellow needle crystal (pure product) at normal temperature, the property is stable, the boiling point is 310-312 ℃, the melting point is 178 ℃, the crystal is insoluble in water, slightly soluble in ethanol and methanol, and soluble in organic solvents such as benzene, toluene, xylene, chloroform, ether, acetone and the like.

In 1933, Cook et al, UK scientist J.W. Cook et al, isolated pure benzopyrene from asphalt for the first time, and proved by animal experiments that benzopyrene can induce skin Cancer in mice, and thus, benzopyrene was identified as the first carcinogen of chemical environment, and long-term exposure to environment containing benzopyrene would undoubtedly cause great harm to human body (see references "Cancer Letters, 2004, 207: 157-" Pulmony Pharmacology & Therapeutics, 2011, 24:133- "and" Journal of Toxicology & Environmental Health Part A, 2003, 66(13):1189- "specifically).

Meanwhile, benzopyrene is widely existed in the environment, and the source of benzopyrene mainly has two aspects: the method is characterized in that waste gas generated by incomplete combustion of fuels such as coal, petroleum and natural gas in the industrial production and living processes, including automobile tail gas, smoke generated in rubber production and smoking, enters food for human life such as vegetables, fruits, grains, aquatic products and meat through pollution to water sources, atmosphere and soil; secondly, in the smoking, baking and frying process of the food, fat, cholesterol, protein, carbohydrate and the like can generate a pyrolysis reaction under a high temperature condition, and polycyclic aromatic hydrocarbon substances including benzopyrene can be formed through cyclization and polymerization reaction, and especially when the food is burnt in the smoking and baking process, the generation amount of the benzopyrene is increased by 10-20 times compared with that of the common food. It can be seen that people are exposed to environments containing benzopyrene almost every moment in daily life. Therefore, there is an urgent need to find a product that is effective in preventing and/or treating benzopyrene exposure.

Disclosure of Invention

[ problem ] to

The invention aims to provide a product capable of preventing and/or treating benzopyrene exposure.

[ solution ]

In order to solve the problems, the invention provides application of lactobacillus plantarum CCFM8661 in preparation of a medicine for preventing and/or treating benzopyrene exposure, wherein the lactobacillus plantarum CCFM8661 is preserved in Guangdong province microorganism strain preservation center with the preservation number of CGMCC No.5494 and the preservation date of 2011, 11 and 29 days.

In one embodiment of the invention, the number of viable bacteria of lactobacillus plantarum CCFM8661 in the medicine is not less than 1 x 10⁶CFU/mL or 1X 10⁶CFU/g。

In one embodiment of the invention, the medicine contains lactobacillus plantarum CCFM8661, a pharmaceutical carrier and/or a pharmaceutical excipient.

In one embodiment of the invention, the drug carrier comprises microcapsules, microspheres, nanoparticles and/or liposomes.

In one embodiment of the present invention, the pharmaceutical excipient comprises an excipient and/or an additive.

In one embodiment of the invention, the excipient comprises a solvent, a propellant, a solubilizer, a cosolvent, an emulsifier, a colorant, an absorbent, a diluent, a flocculant, a deflocculant, a filter aid, and/or a release retardant.

In one embodiment of the invention, the additive comprises microcrystalline cellulose, hydroxypropyl methylcellulose and/or refined lecithin.

In one embodiment of the present invention, the pharmaceutical composition is in the form of powder, granule, capsule, tablet, pill or oral liquid.

The invention also provides a medicine for preventing and/or treating benzopyrene exposure, which contains lactobacillus plantarum CCFM 8661; the lactobacillus plantarum CCFM8661 is preserved in Guangdong province microbial strain collection center, the preservation number is CGMCC No.5494, and the preservation date is 2011, 11 and 29 days.

In one embodiment of the invention, the number of viable bacteria of lactobacillus plantarum CCFM8661 in the medicine is not less than 1 x 10⁶CFU/mL or 1X 10⁶CFU/g。

In one embodiment of the invention, the medicine contains lactobacillus plantarum CCFM8661, a pharmaceutical carrier and/or a pharmaceutical excipient.

In one embodiment of the invention, the drug carrier comprises microcapsules, microspheres, nanoparticles and/or liposomes.

In one embodiment of the present invention, the pharmaceutical excipient comprises an excipient and/or an additive.

In one embodiment of the invention, the excipient comprises a solvent, a propellant, a solubilizer, a cosolvent, an emulsifier, a colorant, an absorbent, a diluent, a flocculant, a deflocculant, a filter aid, and/or a release retardant.

In one embodiment of the invention, the additive comprises microcrystalline cellulose, hydroxypropyl methylcellulose and/or refined lecithin.

In one embodiment of the present invention, the pharmaceutical composition is in the form of powder, granule, capsule, tablet, pill or oral liquid.

The invention also provides application of the lactobacillus plantarum CCFM8661 in removal of benzopyrene, wherein the lactobacillus plantarum CCFM8661 is preserved in Guangdong province microbial strain collection center with the preservation number of CGMCC No.5494 and the preservation date of 2011, 11 and 29 days.

The invention also provides a benzopyrene scavenging agent, which contains lactobacillus plantarum CCFM 8661; the lactobacillus plantarum CCFM8661 is preserved in Guangdong province microbial strain collection center, the preservation number is CGMCC No.5494, and the preservation date is 2011, 11 and 29 days.

Has the advantages that:

1. the invention provides a medicine capable of preventing and/or treating benzopyrene exposure, the effective component of the medicine is Lactobacillus plantarum (Lactobacillus plantarum) CCFM8661, and the Lactobacillus plantarum CCFM8661 can effectively relieve the benzopyrene exposure, and is specifically represented as follows:

(1) the content of benzopyrene metabolite 3-hydroxy benzopyrene in the mouse feces exposed by benzopyrene is remarkably reduced;

(2) the activity of glutamic-oxaloacetic transaminase (AST) and glutamic-pyruvic transaminase (ALT) in the serum of a benzopyrene-exposed mouse is obviously reduced;

(3) the levels of superoxide dismutase (SOD) and Malondialdehyde (MDA) in the brain tissue of the mouse exposed by benzopyrene are obviously reduced;

(4) the behavioral indexes of the pyrene-exposed mice are obviously improved;

(5) the content of short-chain fatty acid in the feces of the mice exposed by benzopyrene is obviously improved.

2. The invention provides a benzopyrene scavenging agent, which can scavenge benzopyrene and is specifically represented by the following components: the effective component of Lactobacillus plantarum (Lactobacillus plantarum) CCFM8661 is added into dimethyl sulfoxide (DMSO) containing benzopyrene to be cultured for 2 hours, so that the clearance of the benzopyrene in the dimethyl sulfoxide (DMSO) can reach more than 60 percent.

Drawings

FIG. 1: clearance rate of benzopyrene in dimethyl sulfoxide (DMSO) by different lactobacillus plantarum.

FIG. 2: the content of benzopyrene metabolite 3-hydroxy benzopyrene in the feces of mice exposed by benzopyrene of different groups.

FIG. 3: activity of glutamic-oxaloacetic transaminase (AST) in sera of mice exposed to benzopyrene of different groups.

FIG. 4: different groups of benzopyrenes expose alanine Aminotransferase (ALT) activity in the serum of mice.

FIG. 5: different groups of benzopyrenes exposed the level of superoxide dismutase (SOD) in mouse brain tissue.

FIG. 6: different groups of benzopyrenes exposed the level of Malondialdehyde (MDA) in mouse brain tissue.

FIG. 7: resting time of different groups of benzopyrene exposed mice.

FIG. 8: total distance of different groups of benzopyrene exposed mice.

FIG. 9: the central zone of the mice was exposed to benzopyrene for the time of activity for the different groups.

FIG. 10: different groups of benzopyrene exposed peripheral zone activation times of mice.

FIG. 11: the content of acetic acid in the mouse feces exposed by benzopyrene of different groups.

FIG. 12: the content of propionic acid in the mouse feces of different groups of benzopyrene exposed.

FIG. 13: the content of isobutyric acid in the mouse feces of the different groups of benzopyrenes exposed.

FIG. 14: the content of butyric acid in the mouse feces exposed by benzopyrene of different groups.

FIG. 15: the content of isovaleric acid in the mouse feces exposed by benzopyrene of different groups.

FIG. 16: the content of valeric acid in the mouse feces of different groups of benzopyrene exposed.

Detailed Description

SPF grade 8 week old male BALB/C mice referred to in the examples below were purchased from Slek laboratory animals, Inc.; skim milk referred to in the examples below was purchased from fumace biotechnology limited; corn oil referred to in the following examples was purchased from alatin Biotechnology Ltd; the skim milk powder referred to in the following examples was purchased from nieuruiz food ltd; the benzopyrene standards referred to in the examples below were purchased from sigma corporation, usa; dimethyl sulfoxide (DMSO) referred to in the following examples was purchased from national pharmaceutical group chemicals, ltd; the Lactobacillus plantarum (Lactobacillus plantarum) CCFM8661 referred to in the following examples has a accession number of CGMCC No.5494, described in the patent application publication No. CN 102586148A; the lactobacillus plantarum (lactobacillus plantarum) CCFM382 referred to in the following examples has a accession number of CGMCC No.9734, and is described in the patent application publication No. CN 104357349A; the Lactobacillus plantarum (Lactobacillus plantarum) CCFM8610 referred to in the examples below has a accession number of CGMCC No.6077, described in the patent application publication No. CN 102827796A; lactobacillus plantarum (Lactobacillus plantarum) Lp45, described in the following examples, has a accession number CGMCC No.8072 and is described in the patent application publication No. CN 110192654A.

The media involved in the following examples are as follows:

MRS solid medium (g/L): 10g/L of peptone, 10g/L of beef extract, 20g/L of glucose, 2g/L of sodium acetate, 5g/L of yeast powder and 2g/L, K of diammonium hydrogen citrate₂PO₄·3H₂O 2.6g/L、MgSO₄·7H₂O 0.1g/L、MnSO₄0.05 g/L, Tween 801mL/L, agar 20g/L, cysteine hydrochloride 0.5g/L, and pH 6.8.

MRS liquid medium (g/L): 10g/L of peptone, 10g/L of beef extract, 20g/L of glucose, 2g/L of sodium acetate, 5g/L of yeast powder and 2g/L, K of diammonium hydrogen citrate₂PO₄·3H₂O 2.6g/L、MgSO₄·7H₂O 0.1g/L、MnSO₄0.05 g/L, Tween 801mL/L, cysteine hydrochloride 0.5g/L, and pH 6.8.

The detection methods referred to in the following examples are as follows:

the detection method of viable count comprises the following steps: the national standard GB 4789.35-2016 food safety national standard food microbiology detection of lactobacillus is adopted.

The preparation of Lactobacillus plantarum cells and cell suspensions referred to in the following examples was as follows:

streaking lactobacillus plantarum liquid on an MRS solid culture medium, and culturing for 48h at 37 ℃ to obtain a single colony; selecting a single colony, inoculating the single colony in an MRS liquid culture medium, culturing for 18h at 37 ℃ for activation, and continuously activating for two generations to obtain an activation solution; inoculating the activated liquid into an MRS liquid culture medium according to the inoculation amount of 2% (v/v), and culturing for 18h at 37 ℃ to obtain a bacterial liquid; centrifuging the bacterial liquid for 15min at 5000g to obtain lactobacillus plantarum thalli; washing Lactobacillus plantarum thallus with phosphate buffer solution with pH of 7.2 for 3 times, and suspending in skimmed milk powder solution with concentration of 130g/L to bacterial concentration of 1.5 × 10¹⁰CFU/mL to obtain bacterial suspension, and storing the bacterial suspension at-80 ℃ for later use.