The Effects Of AP On NMDAR1,NOS,ChAT In Brain Of Morphine Addictive Mouse And Behavioral Activition

Background and objectOpioids medicine are analgesic applied widely in clinics. But opioids easily produce tolerance to body, so as to keep intrinsic effect, drugs must be repeatedly used and given high dose which induce drug dependence. In recent years, extrinsic opioids abuse has been being a severe social problem increasingly. Banning drugs and abstaining from drugs are one of hotspots focused on by the world and one of knotty in medical research.A great deal of experiments related to mechanism of opioids dependence have been put up since the 70s of last century and a series of advances have been reached such as: supersensitivity theory, t metabolism disfigure hypothesis , opioids receptor decreasing theory ,the theory of cAMP system increasing and CREB theory. but ,there has been no clear explanation on the relation of opioids dependence to special neorotransmitters or neuro-peptides.In recent years, excited amino acids and their receptor (mainly NMD A) system and the interaction between the system and opioids receptors system, which participate in molecule mechanism depended on opioids are being given more and more attention. In the process of opioids dependence, excited amino acids receptors (mainly NMDAR) increase complementarity and becomes supersensitive, moreover, resulting in the plastic changing of nerve cell and behavioral activation .Another pointattentioned, in the process of opioids addiction a great deal researches indicated that memory obstacle in which memory submits pathology, namely ,pathologic boosting up . In the process of opioids treated, the whole effect of behavioral activation is impersonal and scientific determinant guideline for drugs and curative effect of abstaining from morphine.Neuropeptides are some important substances which have functions of neuronal signal transmission and regulation. They distribute widely in the central nervous system, and participate in many kinds of functional regulation. An Yuhui isolated some small neuropeptides from bovine brains and found a new peptide: acidic neuropeptidel (AP). It is a small molecular peptide and has no antigenicity. Animal experiments have proved the following effects:? AP can inhibit the exciting signs and get no addiction within experimental chroma; ?ABNP-1 can cure effectively the outbreaking from morphine dependence mice after withdrawal ;(§)AP can increase amino acid of brain and affect the contents of cGMP of brain. In this study, the effects of AP on AD mice were observed by setting up addictive mouse model, in which morphine was injected into abdomen of the mice. The curing mechanism of AP morphine addictive mice was studied by the analysis of the behavioral activation related to learning and memorizing and relavant index in CNS. The purpose of this research is to accumulate valuable data for researching pathogenesis morphine dependence and to explore new medicine for treated of morphine dependence. Materials and MethodsEighty healthy mice,including were randomly divided into 8 groups and each group contained 10 mice In the research, addictive animal model is established by injecting morphine lOOmg/Kg i.p into mice. I group was normal contral group; II group was addictive animal model; III was addictive animal mode+ natural withdrawal , IV was addictive animal model+ orally normal saline,V,VI,VII group was addictive animal model + orally AP80mg/ kg d, BANP-60mg/ kg d, BANP-40mg/ kg d respectively. Morally AP60mg/ kg d.Normal control group and model group mice had not been treated. This treated process lasted for 15days, onceevery day, each timel > 0.5 > 0.25ml respectively. After addictive mice were treated with different dosages of AP, the learning and remembering ability was tested by Y-maze experiment and jumping table experiment at the same time. Withdrawal signs must be observed and recorded carefully .getting out the brain of mice and anaesthesia and perfusion, Immunohistochmistry was used to check the expression of NMDAR1,NOS and ChAT in each group. The experimental data were expressed by and analyzed by one-way analysis of variance (ANOVA). a =0.05 was the standard of test. Result1 The characters of mice' aspect in the course of experiment ?The mice in normal group are good at activity, response and diet, phycological condition; .?In the first 4d there was no remarkable distinction between model group and normal group ,in the subsequent 7d the mouse of model group were scorcher in hair, slower in action, less in diet and cachexia; ?These phenomenon are ameliorated remarkably in high dose treated group compared with model group in behavior and aspect; ?The behavior and aspect of the mouse in NS group, natural withdrawal group, low dose group and middle dose group are similar to model group; ?The mice in only AP group are good at activity, response and diet, even better than normal group.2 the weight change in every group compared with model group ?the mouse weight in AP treated group decreased remarkably especially in the period from day4-ll(p<0.05) while the weight is elevated in only AP group and the weight is remarkably high in the day 1 l(p<0.05); ?compared with natural withdrawal group, the mouse weight in the high dose treated group increased remarkably(p<0.05) ,the mouse weight in the low dose treated group also increased but not remarkably remarkably; ?compared with NS group the mouse weight in the high dose treated group increased remarkably(p<0.05) and so did to some extent in low and middle dose treated group but not remarkably(p>0.05); ?the mouse weight in the high dose treated group are remarkably higher than that in low and middle dose treated groups(p<0.05) and there is no remarkable distinction between low and middle dose group(p>0.05).3 learning ability change through jumping table experiment ?Compared with the normal group the learning ability decreased remarkably in model group(p<0.05), and only AP show the the increase but it is not remarkable(p>0.05); ?Compared with model group the learning ability did not remarkably change in natural withdrawal group and NS group but increased remarkably in high dose treated group. The learning ability of low and middle dose group also increased but not remarkably (p>0.05); ?Compared with natural withdrawal group the learning ability of high dose treated group increased remarkably(p<0.05) and the learning ability of low and middle dose group also increased but not remarkably (p>0.05); ?Compared with NS group the learning ability of high dose treated group increased remarkably and the learning ability of low and middle dose group also increased but not remarkably (p>0.05); ?The learning ability of high dose treated group increased remarkably than the learning ability of low and middle dose group (p<0.05) and there is no remarkable distinction between low and middle dose group(p>0.05).4 Y maze experiment result ?Compared with normal group the learning and memorizing ability through Y maze experiment decreased remarkably in model group(p<0.05), only AP group show the increase but it is not remarkable(p>0.05); (2) compared with model group the learning ability are no change in natural withdrawal group and NS group but increased remarkably in high dose treated group(p<0.05). The learning ability of low and middle dose group also increased but not remarkably(p>0.05); ?Compared with natural withdrawal group the learning ability of high dose treated group increased remarkably(p<0.05) and the learning ability of low and middle dose group also increased but not remarkably(p>0.05); ?Compared with NS group the learning ability of high dose treated group increased remarkably(p<0.05) and the learning ability of low and middle dose group also increased but not remarkably(p>0.05); ?The learning ability of high dose treated group increased remarkably than the learning ability of low and middle dose group (p<0.05),but there is no remarkable distinction between low and middle dose group(p>0.05).5 change of withdrawal signs in the course of AP treatment ? Compared with normal group the withdrawal signs after injecting naloxone in model group like this: weight lose is remarkable, latent period is shorter , times of body stretching are much more(p<0.05), only AP group has no remarkable change(p>0.05); ? Compared with natural withdrawal group withdrawal signs of mice in high dose treated group decreased remarkably (p<0.05)and that of low and middle dose group also decreased but not remarkably(p>0.05); ? Compared with NS group the withdrawal signs of high dose treated group decreased remarkably(p<0.05) and low and middle dose group also decreased but not remarkably(p>0.05); ?The withdrawal signs of high dose treated group decreased remarkably than that of low and middle dose group(p<0.05) ,but there is no remarkable distinction between low and middle dose group(p>0.05).6 the expression of NMDAR1, NOS in hippocampus ?but there is no remarkable distinction between low and middle dose group(p>0.05).Compared with normal group the expression of NMDAR1, NOS in hippocampus increased remarkably in model group(p<0.05), only AP group shows no remarkable change(p>0.05); ?Compared with model group the expression of NMDAR1, NOS dose not change remarkably in natural withdrawal group and NS group (p>0.05)and it decreased remarkably in high dose treated group(p<0.05). The expression of NMDARlin low and middle dose group show some decrease but it is not remarkable(p>0.05); ?Compared with natural withdrawal group the expression of NMDAR1, NOS decreased remarkably in high dose treated group(p<0.05),but in low and middle dose group that showed some decrease but it was not remarkable(p>0.05); ?Compared with NS group the expression of NMDAR1, NOS decreased remarkably in high dose treated group(p<0.05),but in low and middle dose group that showed some decrease but it was not remarkable(p>0.05); (5) the decrease of the expression of NMDAR1, NOS is remarkable in high dose treated group than that in low and middle dose group(p<0.05), but there is no remarkable distinction between low and middle dose group(p>0.05)..7 the expression of ChAT ?Compared with normal group the expression of ChAT decreased remarkably in model group(p<0.05) and there is no remarkable distinction in only AP group(p>0.05); ?Compared with model group the expression of ChAT does not change remarkably in natural withdrawal group and NS group(p>0.05), it increased remarkably in high dose treated group (p<0.05)and not in low and middle dose group even that showed some increase but it was not remarkable(p>0.05); ?Compared with natural withdrawal group the expression of ChAT increased remarkably in high dose treated group(p<0.05),but in low and middle dose group that showed some increase but it was not remarkable(p>0.05); ? Compared with NS group the expression of ChAT increased remarkably in high dose treated group(p<0.05),but in low and middle dose group that showed some increase but it was not remarkable(p>0.05); (5)the decrease of the expression of ChAT is remarkable in high dose treated group than that in low and middle dose group(p<0.05), but there is no remarkable distinction between low and middle dose group(p>0.05).. Conclusions:1. AP could improve the ability of learning and memorizing of addictive mice.2. AP could remarkably increase mouse's weight.3. AP could remarkably inhibit the withdrawal signs of morphine addictive mice.4. the curable effect of Ap on morphine addictive mouse is closely related to dose with experimental dose.5. AP could inhibit the express of NMDAR1,NOS and increase the synthesis of ChAT of morphine addictive mouse's brain .