The Efforts Of The Inflammatory Factors On The Changes Of Neuropeptide S In The Brains Of Asthma Mice Combined With Anxiety

BackgroundBronchial asthma is a kind of physical and mental disease,characterized by chronic airway inflammation and elevated levels of serum inflammatory factors.Asthma can lead to anxiety and anxiety influence asthma control level.But the mechanism of asthma and anxiety comorbidity is still not clear.Recent studies show that,neuropeptide S(NPS)which has anti-anxiety effect and its receptor NPSR1 play a common role in asthma and anxiety disorders comorbidity.On one hand,NPSR1 gene is a susceptibility gene for asthma.On the other hand,the decreased NPS levels in the brain can lead to anxiety.But how to make asthma less NPS in the brain is the key point of researches.This study aims to to investigate the function of inflammatory factors on changes of NPS in the brain to provide a new idea for the study of the comorbidity of asthma and anxiety.PurposeTo investigate the levels of neuropeptide S in the brains of anxiety only mice and asthma mice with anxiety.and the effects of the inflammatory factors on changes of neuropeptide S in cells experiment.Then in cells experiment to investigate the inhibitory effect of the inflammatory factors such as interleukin-1 beta(IL-1?),interleukin-6(IL-6)and tumor necrosis factor-alpha(TNF-?)on the secretion of NPS in neurons,and to study the relationship between he concentration and time of the inhibitory effect of inflammatory factors.Method1.The mice asthma models were constructed by the ovalbumin sensitization method and the anxiety models were established by the uncertainty of the empty bottle water stimulation method.Both methods were used to construct asthma mice with anxiety.Then the mthods of noninvasive pulmonary function testing,BALF eosinophil count and elevated plus maze test were used to validate the models.40 BALB/C mice were randomly divided into 4 groups: the control group,the asthma group,the anxiety group and the asthma and anxiety group.The relative expressions of NPS mRNA in the brain tissue of each group were detected by quantitative real-time polymerase chain reaction(QRT-PCR).2.Rat cortex neurons were got by primary culture and immunofluorescence of MAP-2 markers were used to identify.3.Neurons were divided into 4 groups: the PBS control group,the IL-1? group,the IL-6 group and the TNF-? group.After inflammatory cytokines stimulated neurons,the mRNA expressions of NPS were measured by QRT-PCR and NPS levels in the cell culture fluid were tested by emzyme linked immunosorbent assay(ELISA).4.To stimulate neurons with IL-6 and TNF-? in different concentration(0 ng/L,50 ng/L,100 ng/L,and 200 ng/L),and the neurons were divided into control group,low concentration group,middle concentration group,high concentration group.Then the mRNA expressions of NPS were measured by QRT-PCR and NPS levels in the cell culture fluid were tested by ELISA.5.According to the stimulating time(0 h,6 h,9 h,12 h)of IL-6 and TNF-?,the neurons were divided into 0h group,6h group,9h group and 12 h group.The mRNA expressions of NPS and NPS levels in the cell culture fluid were tested.Results1.The Penh values and BALF Eos% of the the asthma group and the asthma and anxiety group were significantly higher than those of the control group(all P < 0.05).The OT% and OE% values of the anxiety group and the asthma and anxiety group were significantly lower than those of the control group(all P < 0.05).The NPS mRNA levels in brains of the anxiety group and the asthma and anxiety group were significantly lower than the control group(all P < 0.05).The NPS mRNA levels in the asthma and anxiety group were decreased than the anxiety group(P < 0.05).2.Compared with the PBS control group,the mRNA expressions of NPS and NPS levels in the cell culture fluid of the IL-1? group were not statistically significant(all P > 0.05).The mRNA expressions of NPS and NPS levels of the IL-6 group and the TNF-? group were lower than those of the PBS control group(all P < 0.05).The TNF-? group had no significant with the L-6 group(all P > 0.05).3.Among the groups stimulated with IL-6 and TNF-? in different concentration,the results of mRNA expressions of NPS and NPS levels were as follows: The low concentration group,the middle concentration group,and the high concentration group were significantly lower than those of the control group(all P < 0.05).The middle concentration group,and the high concentration group were significantly lower than the low concentration group(all P < 0.05).The high concentration group were significantly lower than the middle concentration group(all P < 0.05).4.Among the groups stimulated with IL-6 and TNF-? in different time,the results of mRNA expressions of NPS and NPS levels were as follows: The 6h group,the 9h group and the 12 h group were significantly lower than those of the 0h group(all P < 0.05).The the 9h group and the 12 h group were significantly lower than the 6h group(all P < 0.05).The 12 h group were significantly lower than the 9h group(all P < 0.05).Conclusion1.The decreases of NPS in the brains may be the reason that asthma mice combine with anxiety.2.Inflammatory cytokines such as IL-6 and TNF-?,can inhibit the secretion of NPS in the cerebral cortex neurons,and this inhibitory effect is enhanced with the increase of the concentration of inflammatory factors and the time of action.3.The cause of the decrease of NPS in the brain of asthma mice with anxiety may be that the inflammatory factor can inhibit the secretion of NPS.