Efekti probiotika (Lactobacillus Rhamnosus) na parodontopatiju kod miševa sa hiperhomocisteinemijom: uloga disbioze
Effects of probiotic (Lactobacillus Rhamnosus) on periodontal disease in mice with hyperhomocysteinemia: the role of dysbiosis
Doktorand
Stanišić, DraganaMentor
Jeremić, NevenaČlanovi komisije
Tyagi, SureshObradović, Radmila
Živković, Vladimir
Nikolić Turnić, Tamara
Metapodaci
Prikaz svih podataka o disertacijiSažetak
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 tre...tirani
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.