皮质酮对原代培养海马神经元中IDO和5-HTT的调控作用  

Regulation of IDO and 5-HTT in primary cultured hippocampal neurons by corticosterone

作　　者：谭瑞[1] 刘任[2] 彭晓凤 TAN Rui;LIU Ren;PENG Xiaofeng(Department of Pharmacy,People′s Hospital of Chongqing Banan District,Chongqing 401320,China;Chongqing Key Laboratory of Biochemistry and Molecular Pharmacology/Department of Pharmacology,Chongqing Medical University,Chongqing 400016,China;Department of Pharmacy,Chongqing Hechuan District People′s Hospital,Chongqing 401520,China)

机构地区：[1]重庆市巴南区人民医院药剂科,重庆401320 [2]重庆医科大学药学院药理教研室/重庆市生物化学与分子药理学重点实验室,重庆400016 [3]重庆市合川区人民医院药剂科,重庆401520

出　　处：《国际检验医学杂志》2021年第17期2076-2079,共4页International Journal of Laboratory Medicine

基　　金：重庆市自然科学基金项目(cstc2017jcyjAX0211)。

摘　　要：目的探讨皮质酮对原代培养海马神经元的吲哚胺2,3-双加氧酶(IDO)和5-HT转运体(5-HTT)的调控作用。方法取怀孕18 d的SD大鼠分离的胎鼠海马神经元进行原代培养。采用MTT比色法检测海马神经元存活率;采用实时荧光定量PCR(qPCR)和Western blot分别检测IDO、5-HTT和脑源性神经营养因子(BDNF)mRNA及蛋白的表达水平。结果皮质酮呈时间和浓度依赖性地降低海马神经元存活率,5μmol/L作用24 h引起海马神经元明显损伤(P<0.05)。同时,IDO mRNA和蛋白表达水平显著升高(P<0.05),BDNF mRNA和蛋白的表达水平则显著降低(P<0.05)。皮质酮的作用可以被其受体拮抗剂米非司酮(Ru-486)逆转(P<0.05),而对5-HTT mRNA和蛋白的表达均没有明显作用(P>0.05)。结论皮质酮通过激动皮质酮受体,上调IDO,降低BDNF表达水平,从而损伤海马神经元;皮质酮的作用可能不是通过5-HTT介导产生的。Objective To investigate the effect of corticosterone on the indoleamine 2,3-dioxygenase(IDO)and 5-HT transporter(5-HTT)in primary cultured hippocampal neurons.Methods Primary culture of hippocampal neurons from fetal rats of the SD rat on the 18th day of pregnancy were performed.MTT colorimetric method was used to detect the survival rate of hippocampal neurons;the mRNA and protein expression of IDO,5-HTT and brain-derived neurotrophic factor(BDNF)were detected by real-time fluorescence quantitative PCR(qPCR)and Western blot,respectively.Results Corticosterone decreased the survival rate of hippocampal neurons in a time-and concentration-dependent manner.The treatment with 5μmol/L corticosterone for 24 h caused significant damage to hippocampal neurons(P<0.05).At the same time,IDO mRNA and protein expression increased significantly(P<0.05),while the expression of BDNF decreased significantly(P<0.05).The effect of corticosterone could be reversed by its receptor antagonist Ru-486(P<0.05),while corticosterone had no significant effect on 5-HTT mRNA and protein expression(P>0.05).Conclusion Corticosterone can stimulate the corticosterone receptor,up-regulates IDO,and reduces the expression of BDNF,thereby damage hippocampal neurons;the effect of corticosterone may not be mediated by 5-HTT.

关 键 词：皮质酮 吲哚胺2 3-双加氧酶 5-HT转运体 脑源性神经营养因子 海马神经元 

分 类 号：R952.6[医药卫生—药学]