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Antigenotossicità e Terapia Probiotica Integrata
1. Dipartimento di Biomedicina e Neuroscienze
Dottorato di Ricerca in Medicina Sperimentale e
Molecolare
Università degli Studi di Palermo
Antigenotossicità e Terapia Probiotica Integrata
Marcello Romeo
2. Carcinoma del Colonretto e Flogosi
Il carcinoma colorettale è una delle patologie tumorali maligne più diffuse nel mondo occidentale.
Come importanza nell’uomo è secondo solo al tumore al polmone, mentre nella donna viene dopo
il tumore al polmone e alla mammella.
La loro causa precisa è sconosciuta, ma hanno importanza, tra gli altri fattori, la dieta, una
predisposizione ereditaria, e alcune patologie definite pre-neoplastiche, come le malattie
infiammatorie croniche dell’intestino (morbo di Crohn e retto-colite ulcerosa) e la poliposi
intestinale.
Il carcinoma colorettale mostra un classico esempio di patogenesi multifattoriale.
Accanto a modificazioni genetiche, è necessario che il tumore acquisisca particolari caratteristiche
fenotipiche che gli consentano di svilupparsi, eludere il sistema immunitario, invadere i tessuti
circostanti, creare nuovi vasi e diffondere a distanza attraverso le vie ematiche e linfatiche.
3. Chaperonine e Genotossicità
Numerosi studi hanno dimostrato che tra le
caratteristiche fenotipiche assunte dalle
cellule tumorali del colon vi è l’aumento
intracellulare delle “Heat Shock Proteins –
HSPs”.
Rodolico V et al, Cell Stress Chaperones 2010
4. LA FLOGOSI DELLA MUCOSA INTESTINALE È IL PRIMO SEGNALE D'ALLARME
G. 10
DELLA MOLTIPLICAZIONE NON FISIOLOGICA DELLA CELLULA
Flogosi e Cancerogenesi
MOLTIPLICAZIONE DIFFERENZIAZIONE APOPTOSI
FLOGOSI FLOGOSI PLUS DEGENERAZIONE TRASFORMAZIONE
COLITE COLITE ULCEROSA COLITE CON DISPLASIA NEOPLASIA INTESTINALE
La flogosi della mucosa rappresenta il primo
segnale d’allarme di una moltiplicazione cellulare
FIG. 11
Alimentazione errata
alterata
Alimentazione scorretta e
5. Espressione di Hsp60 nelle IBD
! !
a. morbo di Crohn b. rettocolite ulcerosa
Nelle malattie infiammatorie croniche
dell’intestino si rilevano elevati livelli di HSP
6. Cell Stress and Chaperones
DOI 10.1007/s12192-010-0196-8
ORIGINAL PAPER
Hsp60 and Hsp10 increase in colon mucosa of Crohn’s
disease and ulcerative colitis
Vito Rodolico & Giovanni Tomasello & Monica Zerilli & Anna Martorana &
Alessandro Pitruzzella & Antonella Marino Gammazza & Sabrina David &
Giovanni Zummo & Provvidenza Damiani & Salvatore Accomando &
Everly Conway de Macario & Alberto J. L. Macario & Francesco Cappello
peronins in inflammatory Society International 25 March
Received: 8 March 2010 / Revised:
# Cell Stress bowel diseases 2010
2010 / Accepted: 26 March 2010
Abstract The purpose of this work was to determine in key factors in the activation of the immune system leading to
colon mucosa of Crohn’s disease (CD) and ulcerative colitis inflammation, we propose that they play a central role in the
(UC) in relapse: a) the levels of the chaperonins Hsp60 and pathogenesis of the two diseases, which, consequently, ought
Hsp10; b) the quantity of inflammatory cells; and c) if the to be studied as chaperonopathies.
levels of chaperonins parallel those of inflammation cells.
Twenty cases of CD and UC and twenty normal controls Keywords Chaperonins . Inflammation . IBD .
(NC) were studied using immunohistochemistry, Western Chaperonopathies . Autoimmunity
blotting and immunofluorescence. Immunohistochemically,
Hsp60 and Hsp10 were increased in both inflammatory
bowel diseases (IBD) compared to NC. These results were Introduction
confirmed by Western blotting. Hsp60 and Hsp10 occurred
in the cytoplasm of epithelial cells in CD and UC but not in Crohn’s disease (CD) and ulcerative colitis (UC) are serious
NC. Hsp60 and Hsp10 co-localised to epithelial cells of pathologies whose etiology and pathogenesis are still
mucosal glands but not always in connective tissue cells of incompletely understood (Bouma and Strober 2003;
lamina propria, where only Hsp60 or, less often, Hsp10 was Kucharzik et al. 2006; Devlin and Panaccione 2010). They
found. Cells typical of inflammation were significantly more are considered inflammatory bowel diseases (IBD) because
abundant in CD and UC than in NC. Since chaperonins are inflammation of the gastrointestinal tract is a major
Vito Rodolico and Giovanni Tomasello contributed equally to the
present work.
V. Rodolico : M. Zerilli : A. Martorana P. Damiani 33
Dipartimento di Patologia Umana, Dipartimento di Medicina Interna,
Università degli Studi di Palermo, Malattie Cardiovascolari e Nefrourologiche,
Palermo, Italy Università degli Studi di Palermo,
Palermo, Italy
G. Tomasello
7. Livelli di Hsp60 nelle tappe della cancerogenesi
a. mucosa normale b. adenoma tubulare
c. adenocarcinoma
Le tappe della cancerogenesi si caratterizzano per un progressivo
aumento dei livelli di HSP
F. Cappello, M. Bellafiore, A. Palma, S. David, V. Marcianò, T. Bartolotta, C. Sciumè, G. Modica, F. Farina, G. Zummo, F. Bucchieri.
60KDa chaperonin (HSP60) is over-expressed during colorectal carcinogenesis. Eur J Histochem 47, 105-10 (2003).
8. Espressione di Hsp60 nel tessuto tumorale colorettale
! ! !
a. invasione nervosa b. invasione vascolare c. area di neoangiogenesi
L’Hsp60 è presente in sede di invasione ematica e nervosa dei
tessuti nonché nei focolai di neoangiogenesi
F. Cappello, S. David, F. Rappa, F. Bucchieri, L. Marasà, T.E. Bartolotta, F. Farina, G. Zummo. The expression of HSP60 and HSP10
in large bowel carcinomas with lymph node metastase. BMC Cancer 5, 139 (2005)
9. Positività di Hsp60 nelle metastasi da carcinoma colorettale
a. metastasi linfonodale b. metastasi ossea
L’Hsp60 è aumentata non solo nel tumore primitivo ma anche
nelle metastasi linfonodali e a distanza del tumore
F. Cappello, S. David, F. Rappa, F. Bucchieri, L. Marasà, T.E. Bartolotta, F. Farina, G. Zummo. The expression of HSP60 and HSP10
in large bowel carcinomas with lymph node metastase. BMC Cancer 5, 139 (2005)
10. HSP 60
Conferisce aumentata resistenza alle cellule
tumorali
I livelli di HSP 60 sono correlati al grading e allo
staging della neoplasia
Elevati livelli di HSP 60 nelle cellule tumorali si
associano ad una prognosi peggiore
Cappello F et al, Biomark Med 2008
16. BB-Pol
FEMS Microbiology Letters 240 (2004) 131–136
www.fems-microbiology.org
Anticancerogenic effect of a novel chiroinositol-containing
polysaccharide from Bifidobacterium bifidum BGN4
Hyun Ju You a, Deok-Kun Oh c, Geun Eog Ji a,b,*
a
Department of Food Science and Nutrition, Seoul National University, Seoul 151-742, Republic of Korea
b
Research Center, BIFIDO Co., Ltd., Seoul 151-818, Republic of Korea
c
Department of Bioscience and Biotechnology, Sejong University, Seoul 143-747, Republic of Korea
Received 17 May 2004; received in revised form 17 September 2004; accepted 17 September 2004
First published online 28 September 2004
Edited by W. Kneifel
Abstract
Strains of bifidobacteria have many health-promotion effects. Whole cells or cytoplasm extracts of Bifidobacterium bifidum
BGN4, isolated from human feces, inhibited the growth of several cancer cell lines. The polysaccharide fraction (BB-pol) extracted
from B. bifidum BGN4 had a novel composition, comprising chiroinositol, rhamnose, glucose, galactose, and ribose. Three human
colon cancer cell lines were treated with BB-pol: HT-29, HCT-116, and Caco-2. Trypan blue exclusion assay and BrdU incorpora-
tion assay showed that BB-pol inhibited the growth of HT-29 and HCT-116 cells but did not inhibit the growth of Caco-2 cells.
Ó 2004 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.
Keywords: Bifidobacterium; Chiroinositol-containing polysaccharide; Colon cancer; Growth inhibition; Probiotics
17. 134
BB-Pol
H.J. You et al. / FEMS Microbiology Letters 240 (2004) 131–136
(a)
600
400
nC
(c)
200
(e)
(d)
(b)
0
0 5 10 15 20 25 30 35 40
Time (min)
Bio-LC analysis composition of the BB-Pol
Fig. 1. Bio-LC analysis of the composition of BB-pol. (a) chiroinositol (26.4%); (b) rhamnose (3.9%); (c) glucose (31.5%); (d) galactose (11.0%); (e)
Chiroinositolo: 26.4%
ribose (23.8%).
Rhamnosio: 3.9%
Glucosio: 31.5%
Galattosio: 11%
HT-29 cells even at a concentration of 10 lg mlÀ1. How- the changed DNA synthesis of treated cells as for
Ribosio: 23.85
3
ever, Caco-2 cells were not noticeably inhibited at con- [ H]-thymidine incorporation assay.
18. BGN4 e Network Citochinico
Clinical Immunology (2007) 123, 30—39
a v a i l a b l e a t w w w. s c i e n c e d i r e c t . c o m
w w w. e l s e v i e r. c o m / l o c a t e / y c l i m
Oral feeding of Bifidobacterium bifidum (BGN4)
prevents CD4 + CD45RB high T cell-mediated
inflammatory bowel disease by inhibition of
disordered T cell activation
Namju Kim a,b , Jun Kunisawa b , Mi-Na Kweon c ,
Geun Eog Ji a,d , Hiroshi Kiyono b,⁎
a
Department of Food and Nutrition, Seoul National University, Seoul, South Korea
b
Division of Mucosal Immunology, Department of Microbiology and Immunology, the Institute of Medical Science,
the University of Tokyo, Tokyo, Japan
c
Mucosal Immunology Section, International Vaccine Institute, Seoul, South Korea
d
Research Center, Bifido Inc., Seoul 151-742, South Korea
19. Citochine tipo Th 1
Il BB-Pol prodotto dal
BGN4 riduce la
produzione delle citochine
pro-infiammatorie
Prevention ofof Th1 cytokine production in the BGN4-fed intestine by the feeding of BGN4. Spleen (A) and distal colon (
Inhibition lymphocyte accumulation in the spleen and large mice. Lymphocytes were isolated from the spleen (A
20. Citochine tipo Th 2
Inhibition of Th1 cytokine production in the BGN4-fed mice. Lymphocytes were isolated from the spleen (A) a
Il BB-Pol prodotto dal BGN4 incrementa la
(B) of the CD4+ CD45RBhigh Tcell-transferred mice fed with BGN4 or skim milk and then cultured for 48 h. Cytokine pro
produzione delle citochine anti-infiammatorie
ture supernatants was measured for the BGN4-fed mice and the skim milk-fed mice groups. Results are shown as me
p < 0.05 versus skim milk-fed mice).
tro study, which may not completely reflect in vivo Discussion
. At least our in vitro study demonstrated that even
ed BGN4 possessed immune modulation effect for Probiotics, as preventive or therapeutic agents agai
rol of T cell cytokine responses. have been shown to be an attractive and alte