Aldo Corsetti

Carmine Merola

Federico Fanti

Monia Perugini

Natalia Battista

Natalia Garcia-Gonzalez

Roberta Prete

English

Archivio della Ricerca - Università degli Studi di Teramo

 Lpb. plantarum IMC513 showed a potential ability to modulate, atthe level of the central nervous system, some ECS components inzebrafish. These findings suggest the use of probiotics as innovative dietarystrategy to counteract the emerging health risk of EDs, which isworth to be further investigated..MODULATION OF THE ENDOCANNABINOID SYSTEM BY PROBIOTIC LACTIPLANTIBACILLUS PLANTARUM IMC513 IN ZEBRAFISH EXPOSED TO ENDOCRINE DISRUPTORSRoberta Prete1, Natalia Garcia-Gonzalez1, Federico Fanti1, Carmine Merola1, Monia Perugini1,  Natalia Battista1, Aldo Corsetti11 Faculty of Bioscience and Technology for Food, Agriculture and Environment, University of Teramo, Teramo, ItalyThis project has received funding from theEuropean Union’s Horizon 2020 research andinnovation programme under the MarieSkłodowska-Curie grant agreement No 713714.1. Perugini M (2016) In: Safety Analysis of Foods of Animal Origin 727-741.2. Zarean M, Keikha M, Poursafa P, Khalighinejad P, Amin M, Kelishadi R (2016) Environ. Sci. Pollut. Res. 23: 24642.3. Garcia-Gonzalez N, Prete R, Perugini M, Merola C, Battista N, Corsetti A. (2020) FEMS Microbiol. Lett., 367: fnaa041.4. Prete R, Tofalo R, Federici E, Ciarrocchi A, Cenci G, Corsetti A (2017) Front. Microbiol. 8:2349.5. Garcia-Gonzalez N, Prete R, Battista N and Corsetti A (2018) Front. Microbiol. 9:2392.6. Prete R, Long SL, Lopez Gallardo A, Gahan CG, Corsetti A, Joyce S (2020) Sci. Rep.10:1165. 7. Prete R, Garcia-Gonzalez N, Di Mattia CD, Battista N, Corsetti A (2020) Sci. Rep.10:16340. Lpb. plantarum IMC513 (109 CFU/g) was administrated by directly adding to thewater freeze-dried cells together with the standard diet. Lpb. plantarum IMC513was daily delivered for six months either alone (probiotic group) or as a combinedtreatment (probiotic + 1mg/kg DnHP group). After 6 months of treatment, all fishes (n= 12/group) were euthanized usingtricaine and then dissected. Total length and wet weight of each fish as well as wet weight of each organ (gutand brain) were measured immediately upon sampling. The presence of key genes of the ECS was tested at the transcriptional level byqRT-PCR analysis in the brain of zebrafish. The gene levels of the toll-like receptor9 (TLR9), a key molecule involved in recognition of microbes in the gut, weremeasured in the intestine samples. The levels of two main endocannabinoids (eCBs), anandamide (AEA) and 2-arachidononyl-glycerol (2-AG), and the eCB-like mediator palmitoylethanolamide(PEA) were detected through UHPLC/MS-MS in zebrafish brain samples.EXPERIMENTAL DESIGNAlthough the statistical analysis did not revealed significant changes, 2-AG and PEA levelsresult increased in zebrafish exposed to DnHP.The levels of AEA; 2-AG and PEA were modulated by the daily intake of Lpb. plantarumIMC513, suggesting the ability of probiotic to interact at central level.MODULATION OF eCBs LEVELS BY IMC513 Values are expressed as mean ± SD (n=12) for each treatment group. No significant differences were found by One-way ANOVA analysis (p>0.05). 1 Samples Lenght (mm) Weight (mm) Brain (g) Gut (g) Control 23.25±1.08 0.212±0.043 0.0044±0.0009 0.0158±0.0035 IMC513 23.79±0.66 0.223±0.036 0.0044±0.0008 0.0141±0.0031 DnHP 23.33±1.17 0.226±0.118 0.0041±0.0007 0.0126±0.0023 IMC513+DnHP 23.08±1.84 0.218±0.066 0.0042±0.0007 0.0165±0.0067 During the 6 months treatment period, no fish died in all the treated groups includingthe control. Both DnHP and probiotic administration did not affect total length, neithertotal body weight nor in terms of brain and gut weight.MORPHOMETRIC DATAThe exposure to DnHP significantly upregulated the mRNA levels of CNR1 and CNR2in the brain, whilst the gene expression of cannabinoid receptors resulteddownregulated in the brain of zebrafish fed with probiotic Lpb. plantarum IMC513.mRNA levels of TLR9 were increased by the 6-months administration of the probioticLpb. plantarum IMC513 with respect to the control samples, showing an upregulationof the intestinal expression of the gene encoding for TLR9 in response to the probioticpresence in the gut. Overall, these results reveal a promising protective role ofprobiotic through a downregulation of the ECS by restoring the gene expression ofECS elements to the control level.EFFECTS OF IMC513 ON THE ECSEnvironmental pollutants, used as plasticizers in food packaging and in thousands of everyday products, have become harmful for their impact on human health. Among them, phthalateshave been recognized as emerging endocrine-disrupting compounds (EDs), that can induce toxic effects leading to different animals and humans health disorders (1), including adysregulation of the endocannabinoid system (ECS).Few studies have been carried out to evaluate the effects in in vivo animal models of di-n-hexyl phthalate (DnHP), commonly used in a wide variety of everyday products for themanufacturing of plastic care products and food packaging (2), whereas no studies have been conducted to determine the effect of DnHP exposure on the ECS of adult zebrafish. Moreover,the interest in dietary interventions (i.e. probiotic bacteria), that are able to positively affect human health, is relevant in the scientific community. Among all the beneficial effects thatprobiotics may have, their ability to counteract the toxicity of environmental chemicals, such as EDs, is also emerging (3).The probiotic strain Lpb. plantarum IMC513 was used as dietary intervention. This strain was previously characterized for several properties, including the antigenotoxic activity, the abilityto survive the gastrointestinal conditions (4) and to interact with intestinal cell models (5-7). In addition, this strain was selected for this experimental trial, based on its ability to inhibit thegenotoxicity of DnHP in a preliminary in vitro study (2).In this perspective, the present study was aimed at evaluating the potential neuroprotective impact of the probiotic strain Lactiplantibacillus plantarum IMC513) on the physiology ofzebrafish exposed to DnHP, with a focus on the modulation of the ECS in the brain.GRAPHICAL ABSTRACTCONCLUDING REMARKSREFERENCESThe experimental procedures on zebrafish were approved by the Committee for Animal Care (OPBA) of Teramo University and by the Italian Ministry of Health (authorization number: 529/2018 -PR). The authors gratefully acknowledge Synbiotec s.r.l. (Camerino, Italy) to kindly provide the probiotic strain Lpb. plantarum IMC513.ACKNOWLEDGEMENTSABSTRACT 

MODULATION OF THE ENDOCANNABINOID SYSTEM BY PROBIOTIC LACTIPLANTIBACILLUS PLANTARUM IMC513 IN ZEBRAFISH EXPOSED TO ENDOCRINE DISRUPTORS

https://research.unite.it/bitstream/11575/117716/1/poster_ID83_Prete%20et%20al.pdf

MODULATION OF THE ENDOCANNABINOID SYSTEM BY PROBIOTIC LACTIPLANTIBACILLUS PLANTARUM IMC513 IN ZEBRAFISH EXPOSED TO ENDOCRINE DISRUPTORS

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