Vpliv probiotika Bidifobacterium breve BR03 na raven provnetnih in protivnetnih citokinov ter kratkoverižnih maščobnih kislin pri otrocih s celiakijo

Klemenak, Martina

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Title:	Vpliv probiotika Bidifobacterium breve BR03 na raven provnetnih in protivnetnih citokinov ter kratkoverižnih maščobnih kislin pri otrocih s celiakijo
Authors:	ID Klemenak, Martina (Author) ID Mičetić Turk, Dušanka (Mentor) More about this mentor... ID Langerholc, Tomaž (Comentor)
Files:	DOK_Klemenak_Martina_2016.pdf (3,51 MB) MD5: 6D41883DCC28414E861B27891508A0CC
Language:	Slovenian
Work type:	Doctoral dissertation
Organization:	MF - Faculty of Medicine
Abstract:	Celiakija je imunsko posredovana sistemska bolezen, ki je posledica uživanja glutena. Edini do sedaj učinkovit način zdravljenja je brezglutenska dieta (BGD), dosledno izvajanje le-te pa predstavlja težavo za mnoge osebe s celiakijo. Pri nekaterih osebah kljub strogi BGD simptomi bolezni vztrajajo in pri 40¬¬–50 % je vnetje še vedno prisotno. V več študijah je bilo ugotovljeno, da imajo osebe s celiakijo neravnovesno črevesno mikrobioto (povečan delež gramnegativnih bakterij in zmanjšan delež bifidobakterij), ki ima pomembno vlogo v patogenezi bolezni. Tudi po uvedbi BGD imajo te osebe še vedno zmanjšan delež bifidobakterij. Določene vrste Bifidobacterium breve so v in vitro študijah delovale protivnetno. Cilj doktorske disertacije je bil ugotoviti vpliv 3-mesečnega jemanja Bifidobacterium breve BR03 in B632 (v nadaljevanju probiotik) na provnetni citokin dejavnik tumorske nekroze alfa (TNF-α), protivnetni citokin interlevkin 10 (IL-10) v serumu in na raven kratkoverižnih maščobnih kislin (KVMK) v blatu pri otrocih s celiakijo na BGD. V raziskavo smo vključili 10 otrok z na novo odkrito aktivno celiakijo, 49 otrok s celiakijo na BGD in 18 zdravih otrok. Otroke s celiakijo na BGD smo randomizirali v dve skupini. Prva skupina je 3 mesece prejemala probiotik, druga skupina pa placebo. Meritve citokinov TNF-α in IL-10 so potekale po navodilih proizvajalca z metodo kemiluminiscence na aparatu Immulite One. Iz vzorcev blata smo s pomočjo visokozmogljivostne tekočinske kromatografije analizirali ocetno, propionsko, masleno in mlečno kislino. Fermentacijski indeks ([ocetna kislina] – ([propionska kislina] + [n-maslena kislina]) / [skupne KVMK]) smo uporabili za prikaz vnetnega količnika KVMK. Pri otrocih s celiakijo na BGD je prišlo po jemanju probiotika do značilnega znižanja TNF-α. Ob kontroli, 3 mesece po koncu jemanja probiotika, se je raven TNF-α ponovno značilno dvignila. Vrednosti IL-10 so bile pri večini otrok pod ravnjo kvantifiacije. Otroci z aktivno celiakijo so imeli značilno višje začetne vrednosti ocetne in propionske kisline v blatu v primerjavi z zdravimi otroki. Otroci s celiakijo na BGD so imeli značilno več propionske kisline v primerjavi z zdravimi otroki. Vrednosti mlečne kisline so bile pri večini otrok pod ravnjo kvantifikacije. Po jemanju probiotika ni prišlo do značilnih sprememb vrednosti KVMK v blatu. Fermentacijski indeks (FI) se je 3 mesece po koncu uživanju probiotika še značilno znižal, kar nakazuje zmanjšanje vnetja na ravni črevesja. Z raziskavo smo dokazali, da sta izbrana probiotična seva Bifidobacterium breve BR03 in B632 delovala protivnetno, saj so se po jemanju le-teh vrednosti TNF-α v serumu značilno znižale in nakazano se je znižal FI, ki ponazarja aktivnost kroničnega, s črevesno floro povezanega vnetja.
Keywords:	celiakija, TNF-α, citokini, kratkoverižne maščobne kisline, črevesna mikrobiota, bifidobakterije
Place of publishing:	Maribor
Year of publishing:	2016
PID:	20.500.12556/DKUM-57863
COBISS.SI-ID:	5807167
NUK URN:	URN:SI:UM:DK:QEPGGHXN
Publication date in DKUM:	09.11.2016
Views:	3100
Downloads:	233
Metadata:
Categories:	MF
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Secondary language

Language:	English
Title:	Influence of probiotic Bifidobacterium breve BR03 administration on the level of pro-inflammatory and anti-inflammatory cytokines and short-chain fatty acids in children with celiac disease
Abstract:	Celiac disease (CD) is an immune-mediated disorder triggered by ingestion of gluten. Gluten-free diet (GFD) has so far proved to be the only effective treatment. Many patients with CD have problems with full compliance to GFD. Symptoms persist in some patients despite strict GFD and in 40¬¬–50 % of them inflammation is still present. Increasing evidence suggests that persons with CD have gut microbiota dysbiosis (increased number of Gram negative bacteria and decreased number of bifidobacteria), which play an important role in the disease pathogenesis. Even after a long-term treatment with GFD patients have a decreased number of bifidobacteria. Some strains of Bifidobacterium breve have shown in vitro anti-inflammatory effect on the gut level. The aim of doctoral dissertation was to investigate the influence of three months administration of Bifidobacterium breve BR03 and B632 (afterwards probiotic) on the pro-inflammatory cytokine tumor necrosis factor alpha (TNF-α), anti-inflammatory cytokine interleukin 10 (IL-10) in serum and on the level of short chain fatty acids (SCFA) in faeces of children with celiac disease on GFD. Clinical study was conducted on 10 children with newly diagnosed active CD, 49 children with CD on GFD and 18 healthy children. Children with CD on GFD were randomized in two groups. The first group received probiotic for three months and the second group was administered placebo. Measurements of cytokines TNF-α and IL-10 was performed according to the manufacturer’s instructions with chemiluminescent immunometric assay in Immulite One. Faecal samples were analyzed for acetic, propionic, butyric and lactic acid with high performance liquid chromatography. Fermentation index ([acetic acid] – ([propionic acid] + [n-butyric acid]) / [total SCFAs]) was used for the assessment of inflammatory status in the gut of patients. TNF-α levels were significantly decreased in children with CD on GFD after receiving probiotics for three months. At checkup, three months after the end of intervention with probiotics, TNF-α significantly increased again. IL-10 levels were below the detection level in most of the children. Children with an active CD had significantly higher baseline levels of acetic and propionic acid in faeces compared to healthy children. Children with CD on GFD had significantly higher baseline levels of propionic acid in faeces compared to the healthy children. Lactic acid levels were below detection level in most of the children. After probiotic administration there was no significant differences in individual SCFA levels in faeces. However, faecal fermentation index was significantly decreased three months after the end of probiotic intervention, suggesting a decrease of inflammation on the gut level. Our study has shown that selected B. breve strains have an anti-inflammatory effect. After administration of the probiotic, TNF-α levels in serum as well as faecal FI decreased, which represents chronical, gut microbiota-mediated inflammation.
Keywords:	celiac disease, TNF-α, cytokines, short chain fatty acids, gut microbiota, bifidobacteria

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