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Rebecca De Smet , Debby Laukens , Martine De Vos , Nico Boon , Guy Brusselle ,
1 4 4 2 3
Background Results
The microbiome plays a crucial role in maintaining intestinal Analysis of the microbiome in smoke- and air-exposed mice revealed that
homeostasis. Disruption of this homeostatic environment microbial community structures changed significantly after smoke
leads to destabilisation of the gut immune system and exposure in all parts of the gut. In addition, the abundance of specific
aberrant immune responses against harmless microbiota, species, in particular Bifidobacterium sp. and Clostridium sp., tended to
which may be involved in the development of Crohn’s disease decrease in response to cigarette smoke in both colonic and ileal samples.
(CD). The most prominent environmental risk factor for CD is (a)
(b)
Methods 0.02026
0.02385
IIa4DC
0.01568
C57BL/6 mice were exposed to cigarette smoke (n = 6) or air (c)
0.02783 0.00818
0.01568
IIb4CC
IIb4DC
(n = 6) according to a well-established protocol. After 24 0.01080
IIa4IL
weeks, the animals were sacrificed and multiple parts of the 0.04145 0.03331
0.01080
0.01021
IIb4IL
(5 cigarettes) 0.02
Figure 1. (a) Taxonomic distribution of species on genus level, based on 454 pyrosequencing. (b and c)
30 min rest NMDS plot and abundance-based Jaccard-based distance tree showing significantly different community
structures between the smoking and non-smoking group. I: non-smoking. II: smoking. DC: distal colon. IL:
ileum. CC: caecum.
7 min cigarette smoke exposure
(5 cigarettes)
A general analysis of mucin expression, using AB/PAS and HID/AB, could
not demonstrate an altered expression of acidic and neutral mucins, nor
30 min rest
changes in sulphated and sialylated mucins after cigarette smoke
exposure.
7 min cigarette smoke exposure (c)
(a) (b)
(5 cigarettes)
30 min rest
AB/PAS HID/AB
The microbial composition was analysed using denaturing
Figure 2. Protein expression of mucins in ileum and distal colon after air and smoke exposure.
gradient gel electrophoresis (DGGE) and 454 pyrosequencing. (a) Quantitative scoring showed no significant differences in general mucin expression. (b) AB/PAS:
Furthermore, the expression of mucins was determined at Acidic mucins are stained in blue, neutral mucins are stained in purple-pink. (c) HID/AB: Sulfomucins are
stained in brown-black, sialomucins are stained in blue.
both the mRNA level (real-time PCR) and the protein level
(Alcian Blue (AB)/Periodic Acid Schiff (PAS), High Iron Diamine However, in the ileum, mRNA expression of MUC2 and MUC3
(HID)/AB staining). significantly increased after cigarette smoke exposure (p = 0,04 and p =
0,03 respectively). In the colon, an increased expression of MUC4 was
Conclusion observed (p = 0,04).
Comparative microbial analysis in mice showed a general shift in the
mucosa-adherent bacterial population, as well as specific changes in
NS NS NS *
NS * * NS
Contact
Liesbeth Allais
Department of Pathology
5Blok A, De Pintelaan 185, 9000 Gent, Belgium
Liesbeth.allais@ugent.be