Figure legend
Fig 1. IN OT improved the DSS-induced abnormal stress-related
behavior of mice. During water avoidance stress exposure, DSS-induced
IBD mice treated with intranasal oxytocin for 7 days showed (A) more
activity, (B) less immobility and (C) more grooming behavior than
untreated mice. IN atosiban reversed IN OT effects in IBD mice(A-C).
Data are presented as means ± SEM. one-way ANOVA, n = 6.
***P<0.001 for main effect: DSS versus normal control;
###P < 0.001 for main effect: IN OT versus without IN OT in
DSS-induced IBD mice;$$$P < 0.001 for main effect: IN
atosiban versus without IN atosiban of DSS-induced IBD mice treated with
IN OT.
Fig 2. Intranasal OT restored DSS-induced hippocampal change of
gene expression. A. Representative Western blots of hippocampal nNOS,
ERK, pERK and OTR.B. Statistical graph of pERK/ERK ratio in different
groups. C. Statistical graph of OTR ratio in different groups. D.
Statistical graph of nNOS ratio in different groups. E. Protein
expression of NO in hippocampus between different groups. F. Protein
expression of BDNF in hippocampus between different groups. Data are
presented as means ± SEM. one-way ANOVA, n =3-6. **P<0.01 and
***P<0.001 for main effect: DSS versus normal control;
#P<0.05,##P<0.01,###P < 0.001 and
####P<0.0001 for main effect: IN OT versus without IN OT
in DSS-induced IBD mice;$P<0.05 and $$$P < 0.001
for main effect: IN atosiban versus without IN atosiban of DSS-induced
IBD mice treated with IN OT.
Fig 3. IN OT
ameliorated DSS-induced intestinal inflammation. weight(A), colon
length (B), disease Activity Score (C), expression of TNF-α (D)and IL-1β
(E), histological Score (F), colon macroscopic damage score (G) was
assessed in different groups. H. Representative image of HE staining in
different groups. Data are presented as means ± SEM. one-way ANOVA, n
=6-10. **P<0.01 and ***P<0.001 for main effect: DSS
versus normal control; #P<0.05 and ###P < 0.001
for main effect: IN OT versus without IN OT in DSS-induced IBD
mice;$P<0.05,$$P<0.01 and
$$$P < 0.001 for main effect: IN atosiban versus without IN
atosiban of DSS-induced IBD mice treated with IN OT.
Fig 4. Intranasal OT reverted the changes induced by DSS in HPA
axis and the sympathetic-adrenal medulla (SAM) axis. CRH in
hypothalamus(A), ACTH(B), Cortisol(C), Catecholamine in serum(D),
Oxytocin in hypothalamus(E), Oxytocin in serum(F), Ach(G) and TNF-α(H)
in spleen were assessed in different groups. Data are presented as means
± SEM. one-way ANOVA, n =4-7. *P<0.05,**P<0.01 and
***P<0.001 for main effect: DSS versus normal control;
#P<0.05,##P<0.01 and ###P < 0.001
for main effect: IN OT versus without IN OT in DSS-induced IBD mice; NS:
Without statistically significant;
$P<0.05,$$P<0.01 and $$$P < 0.001
for main effect: IN atosiban versus without IN atosiban of DSS-induced
IBD mice treated with IN OT.
Fig 5. IN OT
attenuated the DSS-induced changes in macrophage cell, NK cell, and Treg
cell populations in the spleen. Percentages of M1 macrophage (A), M2
macrophage(B), CD3-NKp46+NK cell (C), CD4+ CD25+foxp3+Treg cell(D) in
the spleen. Data are presented as means ± SEM. one-way ANOVA, n =4-6.
**P<0.01 and ***P<0.001 for main effect: DSS versus
normal control; #P<0.05 and ##P<0.01 for main
effect: IN OT versus without IN OT of DSS-induced IBD
mice;$P<0.05 and $$P<0.01 for main effect: IN
atosiban versus without IN atosiban of DSS-induced IBD mice treated with
IN OT.
Fig 6.Splenectomy abolished the
protective effects of IN OT against DSS-induced abnormal stress-related
behavior and intestinal inflammation. A-C. Protective effects of IN OT
on DSS-induced abnormal stress-related behavior was abolished by
splenectomy. weight(D), colon length (E), disease activity score (F),
histological score (G), colon macroscopic damage score (H) was assessed
in different groups. I. Representative image of HE staining in different
groups. Data are presented as means ± SEM. two-way ANOVA, n =5.
**P<0.01 and ***P<0.001 for main effect: DSS versus
normal control; #P<0.05 and ##P<0.01 for main
effect: IN OT versus without IN OT of DSS-induced IBD
mice;$P<0.05 and $$P<0.01 for main effect: IN
atosiban versus without IN atosiban of DSS-induced IBD mice treated with
IN OT.
Fig 7. Possible mechanisms for IN OT improved DSS-induced
abnormal stress-related behavior and intestinal inflammation.Intestinal inflammation “communicated” with brain through systemic
communication, HPA axis and neural communication (vagal and sympathetic
system), which induced psychological disorders. IN OT administration
improved DSS-induced abnormal stress-related behavior and intestinal
inflammation through HPA axis and cholinergic anti-inflammation pathway.