Efekti probiotika (Lactobacillus Rhamnosus) na parodontopatiju kod miševa sa hiperhomocisteinemijom: uloga disbioze

Abstract

Uvod: Lactobacillus rhamnosus (LGG) je gram pozitivni fakultativni anaerob, probiotik, koji ne fermentira saharozu i laktozu. S obzirom na antimikrobna, antiinflamacijska i nekariogena svojstva, LGG bi mogao ponuditi smanjenje rizika, ekonomski prihvatljiv način u prevenciji i lečenju parodontalne bolesti kao i odličnu opciju za postizanje i održavanje parodontološkog zdravlja. Cilj: Cilj ove studije bio je da se ispitaju efekti LGG kod zdravih, miševa sa parodontopatiojom (PD), miševa sa urođenim nedostatkom cistation β-sintetaze (hiperhomocisteinemičnih (HHcy) miševa) i miševa sa disbiozom, kao i uticaj urođenog nedostatka cistation β-sintetaze i disbioze na nastanak PD. Materijal i metode: U studiji bilo je uključeno 48 miševa, 36 miševa divljeg tipa (Wild type, WT, C57BL/6J) i 12 miševa sa urođenim nedostatkom cistation βsintetaze (CBS+/- knock-out heterozigoti, B6.129P2‐Cbstm1Unc/J 002853) koji su razvrstani u sledećih osam grupa: zdravi miševi (WT), miševi tretirani probiotikom LGG, miševi kojima je indukovana PD, miševi kojima je indukovana PD i koji su tretirani probiotikom LGG, CBS+/- miševi, CBS+/- miševi koji su tretirani probiotikom LGG, miševi kojima je indukovana disbioza, miševi kojima je indukovana disbioza i koji su tretirani probiotikom LGG. Tretman LGG podrazumevao je tromesečnu primenu peroralno, u dozi od 2.5×105 CFU dnevno. Nakon tretmana životinje su žrtvovane i sprovedene su mikrobiološke analize fecesa, izmeren je gingivalni protok krvi Laser Doppler-om, radiografska i histomorfometrijska analiza parodontalnog tkiva, zimografijska analiza aktivnosti matriksnih metaloproteinaza, Western blot analiza ekspresije proteina i kvantitativna reakcija lančane polimerizacije u realnom vremenu. Rezultati: LGG primnjen kod navedenih grupa miševa pokazao je antimikrobne i antiinflamacijske efekte, poboljšao oporavak parodontalnog tkiva i doprineo očuvanju zdravog. LGG je poboljšao stanje mikrobiote kod svih ekperimentalnih grupa a time uticao i na poboljšanje PD i pokazao preventivno dejstvo u nastanku iste. Takođe, rezultati su pokazali da disbioza i hiperhomocisteinemija, u našem slučaju kao urođeni poremećaj, potpomažu nastanku PD dok hiperhomocisteinemija, kao individualni poremećaj, indukuje disbiozu. Zaključak: LGG preko kompleksnih mehanizama ostvaruje antimikrobne, antiinflamacijske i preventivne efekte u stanju parodontopatije. LGG doprinosi poboljšanju i očuvanju parodontalnog tkiva miševa. HHcy i disbioza utiču i potpomažu nastanku PD.

Introduction: Lactobacillus rhamnosus (LGG) is a gram positive facultative anaerobic, probiotic, which does not ferment sucrose and lactose. Given its antimicrobial, antiinflammatory and non-cariogenic properties, LGG could offer risk reduction, an inexpensive way to prevent and treat periodontal disease as well as an excellent option for achieving and maintaining periodontal health. Aim: The aim of this study was to examine the effects of LGG in healthy mice with periodontal disease (PD), mice with congenital cystation β-synthetase deficiency (hyperhomocysteinemic (HHcy) mice) and mice with dysbiosis, as well as the effect of congenital β-cystation deficiency synthetases and dysbioses on the formation of PD. Material and methods: The study included 48 mice, 36 wild-type mice (Wild type, WT, C57BL/6J) and 12 mice with congenital cystation β-synthetase deficiency (CBS+/- knock-out heterozygotes, B6.129P2 ‐ Cbstm1Unc/J 002853) classified into the following eight groups: healthy mice (WT), mice treated with LGG probiotics, PD-induced mice, PD-induced mice treated with LGG probiotics, CBS+/- mice, CBS+/- mice treated with the LGG probiotic, mice induced by dysbiosis, mice induced with dysbiosis and treated with the probiotic LGG. LGG treatment required quarterly administration orally, at a dose of 2.5×105 CFU. After treatment, the animals were sacrificed and microbiological analyzes of feces, gingival blood flow was measured Laser Doppler, radiographic and histomorphometric analysis of periodontal tissue zymographic analysis of MMP activity, Western blot analysis of protein expression and quantitative real-time chain polymerization reaction were performed. Results: LGG applied to these groups of mice showed antimicrobial and anti-inflammatory effects, improved periodontal tissue recovery and contributed to maintaining healthy. LGG improved the condition of the microbiota in all experimental groups and thus influenced the improvement of periodontitis and showed a preventive effect in its development. Also, the results showed that dysbiosis and hyperhomocysteinemia, in our case as a congenital disorder, contribute to the development of periodontitis while hyperhomocysteinemia, as an individual disorder, induces dysbiosis. Conclusions: LGG through complex mechanisms achieves antimicrobial, anti-inflammatory and preventive effects in the periodontal disease. LGG contributes to the improvement and preservation of periodontal tissue in mice. HHcy and dysbiosis affect and promote the development of periodontitis.