抑制HDAC4减少JNK/c-Jun活性及其依赖的神经元凋亡  

Inhibiting HDAC4 Reduces JNK/c-Jun Activity-dependent Neuronal Apoptosis

作　　者：曹娅莉 吴力强 刘锶锶 王业忠 袁忠民[1] CAO Ya-li;WU Li-qiang;LIU Si-si;WANG Ye-zhong;YUAN Zhong-min(Department of Neurosurgery//Department of Nuerosurgery Disease Researth Center,The Second Affiliated Hospital of Guangzhou Medical University,Guangzhou 510000,China)

机构地区：[1]广州医科大学附属第二医院神经外科//神经外科疾病研究中心,广东广州510000

出　　处：《中山大学学报（医学版）》2018年第5期669-674,共6页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(8167123);广东省自然科学基金(2015A030313475);广州市科创委基金(201510010180)

摘　　要：【目的】研究抑制组蛋白去乙酰化酶成员HDAC4对SD大鼠小脑颗粒神经元(CGNs)凋亡的影响及机制。【方法】将原代培养6~9 d的CGN分为存活对照组(用含K+浓度为25 mmol/L的培养基,即25 K组)、凋亡组(用含K+浓度为5 mmol/L的培养基,即5 K组)和LMK-235处理组(5 K合并LMK-235处理组),在5 K条件下用不同浓度HDAC4抑制剂LMK-235处理细胞。在转染实验中,将CGN分为25 K+siNC组、5 K+siNC组、5 K+siH-DAC4-1组和5 K+siHDAC4-2组,共转染GFP标记目的细胞。用Western blot法检测p-JNK、JNK、p-c-Jun、c-Jun的表达水平,用Hoechst染色法观察并统计核固缩情况,用免疫荧光检测c-Jun磷酸化水平。【结果】Westernblot结果显示,与5 K组比较,LMK-235处理细胞后JNK、c-Jun磷酸化水平下降。核染色结果显示,与25 K组比较,5 K组核固缩率明显增多(P <0.05);与5 K组比较,LMK-235处理细胞后核固缩率明显减少(P <0.05)。与5K+siNC组比较,小分子干扰HDAC4后,HDAC4的表达下调,c-Jun的磷酸化水平降低,CGNs核固缩率明显减少(P <0.05)。【结论】抑制HDAC4通过下调JNK/c-Jun活性使神经元核固缩率减少,从而抑制神经元凋亡。【Objective】To investigate the effect of inhibiting histone deacetylase 4(HDAC4)on survival and apopto-sis in the cultured cerebellar granule neuron and the mechanisms involved in.【Method】Primary cerebellar granule neu-rons(CGN)isolated from SD rats of 6-9 days were divided into the following groups:25 K group(blank control group),5 K group(apoptosis group)and LMK-235 treatment group(5 K^+LMK-235 treatment group).In the LMK-235 treat-ment group,CGN were treated with different concentrations of LMK-235(HDAC4 inhibitor).For further experiments,CGNs were divided into four groups:25K^+siNC group,5 K^+siNC group,5 K^+siHDAC4-1 group,5 K^+siHDAC4-2 group.GFP was transfected to label CGN.The protein expression of p-JNK,JNK,p-c-Jun and c-Jun were examined by Western blot analysis.Cell nuclear pyknosis were examined by hoechst staining.The phosphorylation level of c-Jun was examined by immunofluorescence.【Result】Compared with 5 K group,the levels of p-JNK and p-c-Jun decreased in the LMK-235 treatment group.The nuclear pyknosis rate of 5K group significantly increased compared with that of 25 K group(P<0.05).The nuclear pyknosis rate of LMK-235 treatment group significantly decreased compared with that of 5 K group(P<0.05).Compared with 25 K^+siNC group,knockdown of HDAC4 by siRNAs led to a remarkable decrease in the phosphorylation level of c-Jun and the nuclear pyknosis rate decreased in both the 5 K^+siHDAC4 group(P<0.05).【Conclusion】Inhibition of HDAC4 significantly decreased the nuclear pyknosis rate of CGN through suppresing the JNK/c-Jun activity.

关 键 词：小脑颗粒神经元凋亡 HDAC4 JNK C-JUN 

分 类 号：R74[医药卫生—神经病学与精神病学]