植物乳杆菌培养上清液中具有促进皮肤慢性伤口愈合和抗致病特性的化合物。

Compounds from Lactobacillus plantarum culture supernatants with potential pro-healing and anti-pathogenic properties in skin chronic wounds.

作者信息

Ramos Alberto N, Sesto Cabral Maria E, Arena Mario E, Arrighi Carlos F, Arroyo Aguilar Abel A, Valdéz Juan C

机构信息

Cátedra de Inmunología, Facultad de Bioquímica, Química, Farmacia y Biotecnología, Instituto de Microbiología, Universidad Nacional de Tucumán , Tucumán , Argentina .

出版信息

Pharm Biol. 2015 Mar;53(3):350-8. doi: 10.3109/13880209.2014.920037. Epub 2014 Oct 27.

DOI:10.3109/13880209.2014.920037

PMID:25347359

Abstract

CONTEXT

It is necessary to advance the field of alternative treatments for chronic wounds that are financially accessible to the least economically developed countries. Previously we demonstrated that topical applications of Lactobacillus plantarum culture supernatants (LAPS) on human-infected chronic wounds reduce the pathogenic bioburden, the amount of necrotic tissue, and the wound area, as well as promote debridement, granulation tissue, and wound healing.

OBJECTIVE

To study LAPS chemically and biologically and to find potential molecules responsible for its pro-healing and anti-pathogenic properties in chronic wounds.

MATERIALS AND METHODS

(1) Chemical analysis: extracts were subjected to a column chromatography and the fractions obtained were studied by GCMS. (2) Quantification: dl-lactic acid (commercial kit), phenolic compounds (Folin-Ciocalteu), H2O2 (micro-titration), and cations (flame photometry). (3) Biological analysis: autoinducers type 2 (AI-2) (Vibrio harveyi BB170 bioassay), DNAase activity (Agar DNAase), and Pseudomonas aeruginosa biofilm inhibition (crystal violet technique).

RESULTS

According to its biological activity, the most significant molecules found by GCMS were the following: antimicrobials (mevalonolactone, 5-methyl-hydantoine, benzoic acid, etc.); surfactants (di-palmitin, distearin, and 1,5-monolinolein); anesthetics (barbituric acid derivatives), and AI-2 precursors (4,5-dihydroxy-2,3-pentanedione and 2-methyl-2,3,3,4-tetrahydroxytetrahydrofurane). Concentrations measured (µg/mL): DL-lactic acid (11.71 ± 1.53) and H2O2 (36 ± 2.0); phenolic compounds (485.2 ± 15.20); sodium (370 ± 17); potassium 920 ± 24); calcium (20 ± 4); and magnesium (15 ± 3). DNAase from LAPS had activity on genomic DNA from PMNs and P. aeruginosa.

DISCUSSION AND CONCLUSION

The molecules and biological activities found in LAPS could explain the observed effects in human chronic wounds.

摘要

背景

有必要推动慢性伤口替代治疗领域的发展，使最不发达国家也能负担得起相关费用。此前我们证明，将植物乳杆菌培养上清液（LAPS）局部应用于人类感染的慢性伤口，可减少致病生物负荷、坏死组织量和伤口面积，并促进清创、肉芽组织形成和伤口愈合。

目的

对LAPS进行化学和生物学研究，找出在慢性伤口中具有促愈合和抗致病特性的潜在分子。

材料与方法

（1）化学分析：提取物进行柱色谱分析，所得馏分用气相色谱-质谱联用仪（GCMS）研究。（2）定量分析：dl-乳酸（商用试剂盒）、酚类化合物（福林-西奥尔特试剂）、过氧化氢（微量滴定法）和阳离子（火焰光度法）。（3）生物学分析：2型自诱导物（AI-2）（哈维氏弧菌BB170生物测定法）、DNA酶活性（琼脂DNA酶）和铜绿假单胞菌生物膜抑制（结晶紫技术）。

结果

根据其生物学活性，通过GCMS发现的最显著分子如下：抗菌剂（甲羟戊酸内酯、5-甲基乙内酰脲、苯甲酸等）；表面活性剂（二棕榈酸甘油酯、二硬脂酸甘油酯和1,5-单亚油酸甘油酯）；麻醉剂（巴比妥酸衍生物）和AI-2前体（4,5-二羟基-2,3-戊二酮和2-甲基-2,3,3,4-四羟基四氢呋喃）。测得的浓度（μg/mL）：dl-乳酸（11.71±1.53）和过氧化氢（36±2.0）；酚类化合物（485.2±15.20）；钠（370±17）；钾（920±24）；钙（20±4）；镁（15±3）。LAPS中的DNA酶对中性粒细胞和铜绿假单胞菌的基因组DNA有活性。