L.Impact On Mk-801,Ceftriaxone And Ru38486in Different Stages Of The Morphine Addiction Memory2.The Effects Of Caffeine-treated U251Human Glioma Cell In Vitro

Objective:Establishment and remaining of drug addiction interaction with cue are the process of learn and memory, glutamate system and glucocorticoid receptors are essential in the formation of acquisition, retrieval, reconsolidation and extinction of reward-based memory. To investigate an effect how to MK-801(noncompetitive receptor antagonist), ceftriaxone(enhanced the activity of glutamate transporter-1) and RU38486(Glucocorticoid receptor antagonist) impact on addiction memory and play a role of treatment, we carried out the following experiments.Method:Conditioned place preference (CPP) is one of comparative maturity models in addiction memory, we used MK-801, ceftriaxone and RU38486in different ways to investigate how they affected the morphine-induced CPP behavior during different phases of addictive memory. Preference scores were contrasted using repeated measures ANOVA to determine the change of acquisition, retrieval and reconsolidation of morphine induced addiction memory. Even more,we studied the above-mentioned drugs induced CPP/CPA behavior, motor activity and sensitization.Result:1) Spending time of morphine group in paired with drug was more than another paired with saline(precondition vs. postcondition, p<0.05).2) Contrasted with control group,small-dose MK-801or ceftriaxone alone show no significant difference on the acquisition,extinction and reinstatement of morphine-induced CPP(p>0.05).3) Contrasted with control group,combined action of MK-801and ceftriaxone indicates significant difference on the acquisition,extinction and reinstatement of morphine-induced CPP(P<0.05).4) Contrasted with control group, the RU38486-treated animals (10,20and30mg/kg) that received morphine showed significant difference on the acquisition,extinction and reinstatement of morphine-induced CPP(P<0.05).5) The result showed no significant difference on RU38486groups (10,20and30mg/kg), small-dose MK-801or ceftriaxone alone and combined action of them contrasted with control group(p>0.05).6) MK-801group alone improved motor activity score contrasted with control group(p<0.05).7)The sensitization score of RU38486-treated group(10,20and30mg/kg) showed notable depression contrasted with control group(p<0.05).Conclusion:1) Morphine can induce CPP in mice or rat.2) Small-dose MK-801(0.05mg/kg, i.p.) or ceftriaxone (25mg/kg. i.p.) alone had no effect on the acquisition,extinction and reinstatement of morphine-induced CPP.3)Pretreatment with a combination of both MK-801(0.05mg/kg) and ceftriaxone (25mg/kg) blocked the acquisition and reinstatement and delayed the extinction of morphine-induced CPP.4) Treatment with RU38486prior to morphine conditioning can attenuate the acquisition, retrieval and reconsolidation of morphine-induced CPP.5) CPP or CPA cann’t be induced by RU38486groups (10,20and30mg/kg), small-dose MK-801or ceftriaxone alone and combined action of them.6) Small-dose MK-801enhanced motor activity,however,a combination of MK-801and ceftriaxone attenuated the enhanced motor activity caused by MK-801administration.7) RU38486(10,20and30mg/kg) could blocked the sensitization behaviors of morphine-induced CPP. In conclusion, these above results suggest that NMDARs, GLT-1and GRs may play an important treatment role in drug addictive memory. This provides some experimental evidences of drug development for the treatment of drug addiction. Objective:Recently, many studies reported that caffeine has anti-cancer effects some extent in several cancer types (including glioma), whereas, the results reported from some large prospective cohorts in other country were different, some showed anti-glioma effects, but others indicated no effects. To investigate the effects of cell viability and apoptosis on caffeine-treated U251human glioma cell line in vitro, we carried out the following experiments.Method:MTT and Annexin V/PI flow cytometry were employed undergoing different caffeine concentrations and time to detect the cell viability and apoptosis on caffeine-treated U251human glioma cell line in vitro.Result:1) Cell viability showed the concentration-dependent downtrend in different concentration caffeine-treated (1,2,4,6,10mM)and different time(24、48、72h). Contrasted with control group,these datum were considered statistically significant(P<0.001).2)During4-10mM caffeine, OD values of cell viability was convergent at24h and48h, and then, In10mM, OD value was basically the same at different time(24、48、72h).3)Apoptosis wasn’t notable to accompany with caffeine-concentration increasing at24h and48h, however, at72h, apoptosis began to rise from2mM, still up to peak (21.55%) in10mM. When caffeine-concentration was more than lOmM, cell death increased and showed a concentration-dependent tendency.Conclusion:1) The cell viability of U251was significantly suppressed by caffeine in vitro.2) Along with prolonging action time of caffeine, the suppressing of the cell viability was enhanced.3) In vitro, the early apoptosis of the caffeine-treated U251cell was later, and caffeine-concentration during2-10mM was effective. Within the scope from2mM to lOmM, the results revealed the increasing for concentration-dependent early apoptosis. However, as caffeine exceeding lOmM, showed a lot of necrotic U251cells rapidly. This study suggest that the caffeine-treated U251cell is suppressed proliferation and observed to promote apoptosis in vitro, it provide experimental evidences for further animal experiments and drug development.