>
网站首页期刊介绍通知公告编 委 会投稿须知电子期刊广告合作联系我们
最新消息:
JNK/MAPK信号通路介导甲基丙二酸血症脑损伤的机制研究
作者:吕书博  张展  赵德华  李晓乐  苏立  罗春伟  孟云  王丽雯 
单位:郑州大学第三附属医院 新生儿筛查中心, 河南 郑州 450000
关键词:甲基丙二酸 神经元 凋亡 c-jun氨基末端激酶 小鼠 
分类号:R338.1
出版年·卷·期(页码):2019·38·第五期(837-842)
摘要:

目的:观察甲基丙二酸对小鼠皮层神经元凋亡及细胞中c-jun氨基末端激酶(JNK)、磷酸化c-jun氨基末端激酶(p-JNK)、c-fos、c-jun蛋白水平的影响,探讨甲基丙二酸血症脑损伤的机制。方法:培养小鼠皮层神经元,甲基丙二酸诱导小鼠皮层神经元,采用流式细胞术检测小鼠皮层神经元凋亡情况。将神经元分为对照组(加入正常培养基)、甲基丙二酸组(加入甲基丙二酸浓度为15 mmol·L-1的培养基)和JNK抑制剂组(加入甲基丙二酸浓度为15 mmol·L-1和SP600125浓度为40 μmol·L-1的培养基),采用蛋白质印迹法检测各组神经元JNK、p-JNK、c-fos、c-jun、B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关X蛋白(Bax)、Caspase 3、cleaved-caspase 3蛋白表达水平。结果:与对照组比较,甲基丙二酸浓度为5、10 mmol·L-1时凋亡神经元比例变化不明显(P>0.05),甲基丙二酸浓度为15 mmol·L-1时凋亡神经元比例明显增加(P<0.05)。甲基丙二酸组神经元p-JNK、c-fos、c-jun蛋白相对表达量高于对照组(P<0.05),JNK蛋白相对表达量和对照组比较差异无统计学意义(P>0.05)。甲基丙二酸组Bax、Caspase 3、cleaved-caspase 3蛋白相对表达量高于对照组(P<0.05),Bcl-2蛋白相对表达量低于对照组(P<0.05);JNK抑制剂组Bax、Caspase 3、cleaved-caspase 3蛋白相对表达量低于甲基丙二酸组(P<0.05),Bcl-2蛋白相对表达量高于甲基丙二酸组(P<0.05)。结论:甲基丙二酸可引起小鼠皮层神经元损伤,其机制可能为甲基丙二酸通过JNK/MAPK通路诱导小鼠皮层神经元凋亡。

Objective: To observe the effect of methylmalonic acidemia on the apoptosis of cortical neurons and the c-jun N-terminal kinase (JNK), phosphorylation c-jun N-terminal kinase (p-JNK), c-fos, c-jun protein levels in cortical neuronal, and to explore the mechanism of brain damage induced by methylmalonic acidemia. Methods: The mouse cortical neuronal cells were cultured. The mouse cortical neuronal cells were induced by methylmalonic acidemia. Flow cytometry was used to detect the apoptosis of mouse cortical neurons. Neuron cells were divided into control group (added to normal medium), methylmalonic acidemia group (added to medium with methylmalonic acidemia concentration of 15 mmol·L-1) and JNK inhibitor group (added to medium with methylmalonic acidemia concentration of 15 mmol·L-1 and SP600125 concentration of 40 μmol·L-1).Western blotting was used to detect the protein levels of JNK, p-JNK, c-fos, c-jun, B-lymphoma-2 (Bcl-2), Bcl-2 related X protein (Bax), Caspase 3 and cleaved-caspase 3 in neurons. Results: Compared with the control group, when the concentration of methylmalonic acidemia were 5 mmol·L-1 and 10 mmol·L-1, the change of the proportion of apoptotic neurons was not obvious (P>0.05), when the concentration of methylmalonic acidemia was 15 mmol·L-1, the proportion of apoptotic neurons increased significantly (P<0.05). The relative expressions of p-JNK, c-fos and c-jun proteins in the neurons of methylmalonic acidemia group were higher than those in the control group (P<0.05). There was no significant difference in the relative expression of JNK protein between the methylmalonic acidemia group and the control group (P>0.05). The relative expressions of Bax,Caspase 3 and cleaved-caspase 3 protein in the methylmalonic acidemia group were higher than those in the control group (P<0.05). The relative expression of Bcl-2 protein in the methylmalonic acidemia group was lower than that in the control group (P<0.05). The relative expressions of Bax, Caspase 3 and cleaved-caspase 3 protein in the JNK inhibitor group were lower than those in the methylmalonic acidemia group (P<0.05), and the relative expression of Bcl-2 protein in the JNK inhibitor group was higher than that in the methylmalonic acidemia group (P<0.05). Conclusion: Methylmalonic acidemia can induce cell damage of mouse cortical neurons. The mechanism may be that methylmalonic acidemia can induce apoptosis of mouse cortical neurons by the JNK/MAPK pathway.

参考文献:

[1] KTENA Y P,RAMSTAD T,BAKER E H,et al.Propofol administration in patients with methylmalonic acidemia and intracellular cobalamin metabolism disorders:a review of theoretical concerns and clinical experiences in 28 patients[J].J Inherit Metab Dis,2015,38(5):847-853.
[2] SUI X,KONG N,YE L,et al.p38 and JNK MAPK pathways control the balance of apoptosis and autophagy in response to chemotherapeutic agents[J].Cancer Lett,2014,344(2):174-179.
[3] 李德渊,伍金林,罗黎力,等.c-Jun氨基末端激酶介导的FOXO3a核转位在缺氧缺血性脑损伤新生大鼠神经元凋亡中的作用[J].中国当代儿科杂志,2017,19(4):458-462.
[4] MARTINEZ-ALVAREZ L,JAMESON E,PARRY N R,et al.Optic neuropathy in methylmalonic acidemia and propionic acidemia[J].Br J Ophthalmol,2016,100(1):98-104.
[5] 葛娟,乔凌燕,李堂.甲基丙二酸血症发病和诊治及预后的研究进展[J].中国儿童保健杂志,2017,25(3):258-260,264.
[6] COFFEY E T.Nuclear and cytosolic JNK signalling in neurons[J].Nat Rev Neurosci,2014,15(5):285-299.
[7] 王小虎,吴小涛.IL-1β通过p38、JNK和NF-κB信号通路促进髓核细胞增殖[J].东南大学学报(医学版),2016,35(1):32-36.
[8] 朱翔,曹苏,秦毅彬,等.吡那地尔对术后持续性疼痛大鼠脊髓JNK/MCP-1释放的影响[J].东南大学学报(医学版),2016,35(5):678-682.
[9] 蔡艳,蒋伟,周爱玲,等.苦参素通过p38/JNK信号途径对海马神经元凋亡的影响[J].中国中药杂志,2017,42(4):731-738.
[10] MOLLEREAU B,MA D.Rb-mediated apoptosis or proliferation:It's up to JNK[J].Cell Cycle,2016,15(1):11-12.
[11] ZHAO Y,LUO A,LI S,et al.Inhibitor of differentiation/DNA binding 1(ID1) inhibits etoposide-induced apoptosis in a c-jun/c-Fos-dependent manner[J].J Biol Chem,2016,291(13):6831-6842.
[12] BAI L,MAO R,WANG J,et al.ERK1/2 promoted proliferation and inhibited apoptosis of human cervical cancer cells and regulated the expression of c-Fos and c-Jun proteins[J].Med Oncol,2015,32(3):57.
[13] SZYMANSKA E,SKOWRONEK A,MIACZYNSKA M.Impaired dynamin 2 function leads to increased AP-1 transcriptional activity through the JNK/c-Jun pathway[J].Cell Signal,2016,28(1):160-171.
[14] FAN X,XI H,ZHANG Z,et al.Germ cell apoptosis and expression of Bcl-2 and Bax in porcine testis under normal and heat stress conditions[J].Acta Histochem,2017,119(3):198-204.
[15] LIU X,DONG J,CAI W,et al.The effect of thymoquinone on apoptosis of SK-OV-3 ovarian cancer cell by regulation of Bcl-2 and Bax[J].Int J Gynecol Cancer,2017,27(8):1596-1601.
[16] LIN C H,WU M R,LI C H,et al.Editor's Highlight:Periodic exposure to smartphone-mimic low-luminance blue light induces retina damage through Bcl-2/BAX-dependent apoptosis[J].Toxicol Sci,2017,157(1):196-210.
[17] BHARDWAJ R,SANYAL S N,VAIPHEI K,et al.Sesamol induces apoptosis by altering expression of Bcl-2 and Bax proteins and modifies skin tumor development in Balb/c mice[J].Anticancer Agents Med Chem,2017,17(5):726-733.
[18] LIU L S,BAI X Q,GAO Y,et al.PCSK9 promotes oxLDL-induced PC12 cell apoptosis through the Bcl-2/Bax-caspase 9/3 signaling pathway[J].J Alzheimers Dis,2017,57(3):723-734.

服务与反馈:
文章下载】【发表评论】【查看评论】【加入收藏
提示:您还未登录,请登录!点此登录
您是第 237989 位访问者


copyright ©《东南大学学报(医学版)》编辑部
联系电话:025-87232481 83272483
电子邮件:
bjb@pub.seu.edu.cn

苏ICP备09058364