吗啡对原代培养海马神经元内Gi_2蛋白水平的影响  

Effects of morphine on Gi_2 protein in primary cultured hippocampal neurons

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作  者:吴青华[1] 傅强[2] 王新华[3] 刘立伟[4] 赵建华[1] 唐时荣[1] 

机构地区:[1]解放军第150医院麻醉科,洛阳 471031 [2]解放军总医院麻醉科 [3]上海同济大学附属东方医院麻醉科 [4]第二军医大学长征医院麻醉科

出  处:《解放军医学杂志》2006年第9期879-880,共2页Medical Journal of Chinese People's Liberation Army

基  金:国家自然科学基金资助项目(39870757)

摘  要:目的观察吗啡对原代培养海马神经元内Ⅱ型抑制性鸟苷酸结合蛋白(Gi2蛋白)水平的影响。方法取培养7天的成熟海马神经元,随机分为吗啡处理4h组(M4)、8h组(M8)、16h组(M16)、24h组(M24)、48h组(M48)和空白对照组(C),每组6孔。吗啡处理组加入终浓度为10μmol/L的吗啡,C组加入生理盐水。免疫荧光技术检测神经元内的Gi2蛋白水平。结果与C组比较,M16组、M24组和M48组海马神经元内Gi2蛋白水平显著降低(P<0·01),而M4和M8组无明显变化(P>0·05)。M48组海马神经元内Gi2水平明显低于M16、M24组(P<0·05)。结论吗啡可降低原代培养的海马神经元内Gi2蛋白水平,降低程度与吗啡处理时间密切相关。Objective The molecular basis for opiate tolerance and dependence remains poorly understood despite extensive investigation in several preparations, including the hippocampus. Recent studies have implicated that the hippocampus played a central role in opiate tolerance, dependence and withdrawal. The current study is to explore the change in guanine nucleotide binding protein-inhabitant (Gi2) protein in primary cultured hippocampal neurons with morphine treament. Methods The hippocampus was harvested from newborn Spra-gue-Dawley rats. Primary hippocampal neuronal cultures of 7 days in vitro were used and divided randomly into six groups (n=6), i.e. morphine treatment 4h group (M4), 8h group (M8), 16h group (M16), 24h group (M24), 48h group (M48) and control group (C). All morphine treatment groups were treated with morphine (10μmol/L). C group was treated with saline. The G protein levels were determined with immtmofluorscence and laser scanning confocal microscope (LSCM) imaging techniques. Results Gi2 protein levels in M16, M24 and M48 groups decreased significantly compared with that in C group (P〈0. 01), while Gi2 protein in M4 and M8 groups showed no change (P〉0. 05). Among M16, M24 and M48 groups, Giz protein level was lowest in the M48 group. Conclusion The results indicated that Gi2 protein levels decreased significantly in primary cultured hippocampal neurons with morphine treatment, which might be a potential molecular mechanism of opioid tolerance and dependence.

关 键 词:吗啡 海马 神经元 荧光免疫测定 抑制性鸟苷酸结合蛋白 

分 类 号:R614.1[医药卫生—麻醉学]

 

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