Ghrelin对原代培养大鼠海马神经元内Ca^(2+)浓度影响  

THE EFFECT OF GHRELIN ON CYTOPLASMIC Ca^(2+) OF PRIMARY CULTURED RAT HIPPOCAMPAL NEURONS

作　　者：杨洁[1] 于明[1] 刘帅[1] 周宇[1] 

机构地区：[1]青岛大学医学院生理学教研室,山东青岛266071

出　　处：《青岛大学医学院学报》2017年第2期140-142,145,共4页Acta Academiae Medicinae Qingdao Universitatis

基　　金：国家自然科学基金资助项目(31471079)

摘　　要：目的探讨胃促生长素(ghrelin)对SD大鼠海马神经元细胞内Ca^(2+)浓度影响及可能的机制。方法原代培养的SD大鼠海马神经元细胞,随机分为空白对照组(A组)、4nmol/L ghrelin组(B组)以及YIL718+4nmol/L ghrelin组(C组)。A组细胞不做任何特殊处理;B组细胞在大鼠海马培养14d后,用含有4nmol/L ghrelin的培养液继续孵育24h;C组细胞在4nmol/L ghrelin处理大鼠海马神经元0.5h前,加入10μmol/L的YIL718进行处理。然后采用confocal结合Fluo-4AM荧光钙成像技术实时记录海马神经元细胞内Ca^(2+)浓度及谷氨酸诱导的胞内钙Ca^(2+)浓度。同时结合实时定量反转录聚合酶链反应技术对ghrelin可能引起的基因表达的变化进行分析。结果与A组相比,4nmol/L ghrelin处理24h可升高胞内Ca^(2+)的水平(F=5.671,q=4.489,P<0.01),但可抑制谷氨酸诱导的细胞内Ca^(2+)水平的升高(F=19.770,q=8.038,P<0.001);YIL718预处理可消除ghrelin的影响(q=3.180,P<0.05)。Ghrelin可抑制海马神经元kcnj2mRNA的表达(t=4.101,P<0.05)。结论ghrelin可能通过激活GHS-R1a调节海马神经元细胞内游离Ca^(2+)的浓度,提示ghrelin/GHS-R1a通路对海马神经元的活动有调控作用。Ghrelin抑制谷氨酸诱导的神经元细胞内Ca^(2+)浓度升高,可能是其抑制海马依赖性记忆形成的细胞基础。Objective To explore the effect of ghrelin on cytoplasmic Ca^2＋of primary cultured rat hippocampal neurons and its possible mechanism. Methods Primary cultured hippocampal neurons of SD rats were divided into group A（control group）,group B（4nmol/L ghrelin group）and group C（YIL718＋4nmol/L ghrelin group）at random.The cells in group A were not treated with any special treatment;the cells in group B were treated with 4nmol/L ghrelin for 24 hafter primary cultured at hippocampal neurons for 14d;that in group C were pretreated with 10μmol/L YIL718 for 0.5hbefore 4nmol/L ghrelin treatment.In combination with Fluo-4AM calcium imaging and confocal techniques,baseline intracellular Ca^2＋and glutamate-evoked Ca^2＋increase in cultured hippocampal neurons were recorded.Meanwhile,mRNA expression of interested genes induced by ghrelin treatment was analyzed using real-time qRT-PCR technique. Results Treated with 4nmol/L ghrelin for 24 hcould increase the level of cytoplasmic Ca^2＋（F=5.671,q=4.489,P〈0.01）,but suppress the elevation of glutamate-evoked Ca^2＋（F=19.770,q=8.038,P〈0.001）.Pre-treatment with YIL718 could elimate the impact of ghrelin（q=3.180,P〈0.05）.Ghrelin could inhibit the mRNA expression of kcnj2 in hippocampal neurons（t=4.101,P〈0.05）. Conclusion Ghrelin regulates intracellular free Ca^2＋concentration in hippocampal neurons by activating GHS-R1 aindicates that ghrelin/GHS-R1 apathway plays a regulatory on the activity of hippocampal neurons.Ghrelin inhibits glutamate-induced increase of intracellular Ca^2＋concentration may be the cellular basis for its inhibition of hippocampus dependent memory formation.

关 键 词：胃促生长素 CA3区 海马 神经元 原代培养 钙成像 CA^2+ 

分 类 号：R338[医药卫生—人体生理学]