Study On The Influence Of Different Concentration Gradient Neuromedin B Receptor Agonist To Mice's Gestational Age And Its Mechanism

Object:To observe the influence of different concentration gradient neuromedion B receptor agonist on gestational age and delivery interval of pregnant mice.Study the mechanism of nuclea factorκB—heat shock protein 70,interlukin-6 passway participating in the labor on set.Methods:1.Establish patterns of pregnant mice the gestational age of which is precise and divide them into five groups randomly which are low-dose NMB group,middle-dose NMB group and high-dose NMB group, positive contrast oxytocin group and negative contrast physiological saline group was respectively injected by NMB30μg/kg,90μg/kg,150μg/kg,oxytocin 50mIU/kg and equal amount physiological saline.2.Observe the influence of different grade dosesagonist NMBR on gestational age and delivery interval of Mus musculus.3.Harnessing the analytical method of transcription factor Noshift detects the influence of different grade doses agonist NMBR on binding activity of NF-κB—P65 DNA in myomitrium of pregnant mice.4.Using semi-quantitative RT-PCR and technology Western blotting measures the influence of different grade dosesagonist NMBR on mRNA and protein expression level of IL-6 and HSP70 in myomitrium of pregnant mice.Results:1.The influence of different grade dosesagonist NMBR on gestational age and delivery interval of pregnant mice.1)The gestational age of low-dose NMB group,middle-dose NMB group and high-dose NMB group,positive contrast oxytocin group and negative contrast physiological saline group was respectively 19.88±0.61,19.56±0.57,19.31±0.23,19.32±0.38和19.90±0.50days. Clearly seen that,High-dose NMB group and positive contrast oxytocin group give the shortest time of gestation.The next is middle-dose NMB group,and low-dose NMB group and negative contrast physiological saline group give the longest time of gestation.Compared with low-dose NMB group and negative contrast physiological saline group,The gestational age of High-dose NMB group and positive contrast oxytocin group were shortened significantly which also have statistical significance(P＜0.05).High-dose NMB group and low-dose NMB group doesn't have notable difference(P＞0.05).2)The modified gestational age of low-dose NMB group, middle-dose NMB group and high-dose NMB group,positive contrast oxytocin group and negative contrast physiological saline group is respectively 21.12±14.64,13.50±13.65,3.27±5.48,7.58±9.13和21.45±11.9 mininues.The modified gestational age of high-dose NMB group and positive contrast oxytocin group give the shortest time.The next is middle-dose NMB group,and low-dose NMB group and negative contrast physiological saline group give the longest time of modified gestational age.Compared with low-dose NMB group and negative contrast physiological saline group,The modified gestational age of high-dose NMB group and positive contrast oxytocin group were shortened significantly which also have statistical significance(P＜0.05). High-dose NMB group and low-dose NMB group doesn't have notable difference(P＞0.05).3)The delivery interval of low-dose NMB group,middle-dose NMB group and high-dose NMB group,positive contrast oxytocin group and negative contrast physiological saline group is respectively 19.86±11.88,18.57±9.13,16.17±5.78,17.00±2.52和21.78±8.54minutes. Comparing the delivery interval of pregnant mice in each group,we find all of them don't have notable difference(P＞0.05).2.The mechanism about gestational age of pregnant mice affected byagonist NMBR with different concentration gradient.1)the expression of binding activity of NF-κB—P65 DNA in myomitrium of pregnant mice.Low-dose NMB group,middle-dose NMB group and high-dose NMB group,positive contrast oxytocin group and negative contrast physiological saline group is respectively 0.126±0.055,0.231±0.088, 0.394±0.471,0.202±0.157和0.266±0.103.The expression level of high-dose NMB group is highest.But using analysis OneWay-ANOVA, the difference between high-dose NMB group and other four groups doesn't have statistical significance(P＞0.05).Using independent sample t test,it can be clearly seen that,high-dose NMB group compared with low-dose NMB group has statistical significance(P＜0.05).Positive contrast oxytocin group doesn't present high expression level like high-dose NMB group,But there are no tatistical significance between them(P＞0.05).2)the expression of HSP70 mRNA in myomitrium of pregnant mice is respectively 0.287±0.082,0.286±0.146,0.702±0.298,0.245±0.120和0.308±0.146 in low-dose NMB group,middle-dose NMB group and high-dose NMB group,positive contrast oxytocin group and negative contrast physiological saline group.The relative expression level of high-dose NMB group is obviously superior to other four groups;the differences of them all have statistical significance.3)the expression of protein HSP70 in myomitrium of pregnant mice is respectively 0.471±0.204,0.529±0.185,0.647±0.140,0.324±0.141 and 0.410±0.233 in low-dose NMB group,middle-dose NMB group and high-dose NMB group,positive contrast oxytocin group and negative contrast physiological saline group.The expression of protein HSP70 in high-dose NMB group is obviously superior than that of positive contrast oxytocin group and negative contrast physiological saline group,and the differences between them all have statistical significance(P＜0.05).the expressions of NMB among the three concentration gradient don't have significant difference(P＞0.05).4)The expression of IL-6 mRNA in myomitrium of pregnant mice is respectively 0.150±0.137,0.181±0.252,0.419±0.199,0.559±0.182 and 0.133±0.184 in low-dose NMB group,middle-dose NMB group and high-dose NMB group,positive contrast oxytocin group and negative contrast physiological saline group.Positive contrast oxytocin group and high-dose NMB group all present high expression level,and obviously higher than negative contrast physiological saline group middle-dose NMB group and low-dose NMB group.The differences between them have statistical significance(P＜0.05).the difference between high-dose NMB group and positive contrast oxytocin group doesn't have significant difference(P＞0.05).5)The expression of protein IL-6 in myomitrium of pregnant mice is respectively 0.206±0.206,0.142±0.149,0.118±0.124,0.198±0.229 and 0.086±0.099in low-dose NMB group,middle-dose NMB group and high-dose NMB group,positive contrast oxytocin group and negative contrast physiological saline group,the relative expression of protein IL-6 in the three groups with different concentration gradient ofagonist NMBR and positive contrast oxytocin group expression are all higher than negative contrast physiological saline group,but comparing the five groups,the differences among each group don't have statistical significance(P＞0.05).Conclusion:1.High-concentration NMBR agonist can shorten the gestational age of pregnant mice efficiently.2.NMBR agonist affects gestational age through up-regulation NF-kB—IL-6,HSP70 expression.Maybe,there are other transcription ways or transcription factor of NF-kB involed in signal transduction pathways.