The sympathetic mesenteric nerve regulates experimental colitis
Rose A
1
Willemze ,
Sigrid EM
1
Heinsbroek ,
Laurens EJ
1
Nijhuis ,
1
Welting ,
Olaf
Joost HA
2
Folgering ,
Harold
2
Darwinkel ,
Daniel
3
Chew ,
Sonal
3
Patel ,
Wouter J de
1
Jonge
1. Tytgat Institute for Liver and Intestinal Research, Academic Medical Center, Amsterdam, The Netherlands 2. Brains On-line B.V., Groningen, The Netherlands 3. GlaxoSmithKline, Middlesex, United Kingdom
Contact: [email protected]
Funded by: GSK Bioelectronics
Methods
Background & Aim
high
CD45RB
1)
The autonomous nervous system plays a regulatory role in the immune response.
T-cell transfer colitis was induced in
mice to
investigate the effect of chemical sympathectomy, using 6hydroxydopamine (6-OHDA), or surgical sympathectomy, cutting the
supra mesenteric nerve (Sx).
The sympathetic neurotransmitter norepinephrine has anti-inflammatory effects on
1
dendritic cell activation. This is a critical event in colitis.
Our aim was to investigate the effect of norepinephrine depletion via surgical or
chemical sympathectomy on the course of experimental colitis.
-/Rag1
2) In a dextran sodium sulphate (DSS)-induced colitis model, Sx or Sx
combined with gastrointestinal-selective vagotomy (Cx) was
performed.
1. Nijhuis, L.E. et al. Adrenergic beta2 receptor activation stimulates anti-inflammatory properties of dendritic cells in vitro. PloS one 9, 2014.
Results
Weight
IL-6
200
0
6-OHDA
Vehicle
6-OHDA
0
Transferred
1000
500
0
Sx
Fig. 3 Ileal norepinephrine levels
110
400
105
100
* ** ** ** **
* *
95
90
Sx
-
Sx
**
300
200
100
0
85
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
*
**
8
100
Sx
Cx
DSS
DSS + Sx
DSS + Cx
95
90
85
1
Sx
2
3
4
*
6
4
2
0
5
6
7
-
8
Days
Sx
Cx
-
Sx
Vehicle
Cx
DSS
Fig. 6 Clinical parameters for colitis (weight loss and and colon lenght). Statisical analysis was
performed with Mann-Withney U test * P < 0.05, ** P < 0.01
IL-1
Colon weight/6cm
Weight (mg)
***
0
Fig. 2 Histology score after CD45RBhigh T-cell
transfer and 6-OHDA treatment
Weight
Norepinephrine
1500
Non-transferred
-
Sx
2
-
Sx
Days
Fig. 4 Clinical signs of colitis (weight loss and colon weight)
Statistical analysis was performed with Mann-Withney U test * P < 0.05, ** P < 0.01
IL-10
6
4
*
2
0
-
IL-12
Sx
6
4
2
0
-
Sx
IL-6
M CP-1
Relative mRNA expression
Fig. 1 Ileal norepinephrine levels
-
0
Relative mRNA expression
Vehicle 6-OHDA
0
2
**
4
**
10
105
6
4
2
*
60
40
20
0
-
0
-
Sx
Fig 5. mRNA expression of different inflammatory cytokines in the colon, relative
to the sham-operated mice, normalized for housekeeping genes.
Statistical analysis was performed with Mann-Whitney U test. * P < 0.05,
** P < 0.01, *** P < 0.001
Relative mRNA expression
Vehicle
2
4
Colon length
**
Length (cm)
5
***
6
Change in bodyweight %
10
4
Relative mRNA expression
400
p = 0.09
6
Relative mRNA expression
600
6
Relative mRNA expression
Relative mRNA expression
15
Score
pmol/gr tissue
*
-
IL-1 
TNF-
20
Change in bodyweight %
In DSS-induced colitis, Sx did not
significantly affect colonic IL-1β and IL-6
expression, although Cx did augment
clinical signs and biomarkers compared to
sham-operated mice in DSS-induced colitis
in the distal colon (Fig. 6 and 7).
Histology score
800
pmol/gr tissue
When Sx was performed, a significant
decrease of norepinephrine was observed
(Fig. 3). Furthermore, Sx caused clinical
signs of spontaneous colitis starting at day
8 (Fig. 4) and a significant increase of proinflammatory cytokines IL-1β and IL-6 (Fig.
5). No spontaneous colitis was seen after
Sx in WT mice (not shown).
Norepinephrine
Relative mRNA expression
In CD45RBhigh T-cell transfer colitis, 6OHDA caused a significant decrease of
norepinephrine (Fig. 1), but had no effect
on colitis parameters (not shown).
Surprisingly however, in control Rag1-/mice, 6-OHDA aggravated colitis pathology
score, however not significantly (Fig. 2).
Sx
Vehicle
Cx
-
Sx
DSS
Cx
200
**
150
100
50
0
-
Sx
Cx
-
Vehicle
Fig. 7 mRNA expression of different inflammatory cytokines in the distal colon, relative to the
vehicle-treated, sham-operated mice, normalized for housekeeping genes. Statistical analysis with
Mann-Whitney U test. * P < 0.05, ** P < 0.01
Conclusion
Sympathetic innervation maintains homeostasis in the gut.
Norepinephrine is critical in maintaining mucosal tolerance towards luminal microbiota. This may
explain how neuronal damage and plasticity contributes to pathology in IBD.
Sx
DSS
Cx