短暂前脑缺血再灌注对大鼠海马中脑源性神经营养因子启动子与组蛋白去乙酰化酶3结合的影响及其作用机制  

Effect of transient forebrain ischemia-reperfusion on the binding of brain-derived neurotrophic factor promoters to histone deacetylase 3 in hippocampus of rat and its mechanism

作　　者：张倩[1] 严德萍 史晋朝 钟锦 周阳 赵欣[1] 张宇[1] 李建国[1] ZHANG Qian;YAN De-ping;SHI Jin-chao;ZHONG Jin;ZHOU Yang;ZHAO Xin;ZHANG Yu;LI Jian-guo(Key Laboratory of Cellular Physiology,Ministry of Education,Department of Physiology,Shanxi Medical University,Taiyuan 030001,Shanxi Province,China)

机构地区：[1]山西医科大学生理学系细胞生理学教育部重点实验室,山西太原030001

出　　处：《中国生物制品学杂志》2023年第2期133-137,共5页Chinese Journal of Biologicals

基　　金：国家自然科学基金(81671231);山西省回国留学人员基金(2017-056);山西省“1331工程”重点学科建设计划(XK201708)。

摘　　要：目的评价短暂前脑缺血再灌注(ischemia-reperfusion,I/R)对大鼠海马中脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)启动子与组蛋白去乙酰化酶3(histone deacetylase 3,HDAC3)结合的影响,并探讨其作用机制。方法采用Pulsinelli四血管夹闭法建立SD大鼠的I/R模型(I/R组),同时设假手术组(Sham组)。尼氏染色法观察大鼠海马中神经元存活情况;染色质免疫共沉淀(chromatin immunoprecipitation,ChIP)法检测大鼠海马中BDNF启动子(Bdnf-p1、Bdnf-p2、Bdnf-p4和Bdnf-p6)与HDAC3的结合情况;qPCR法检测大鼠海马中反义脑源性神经营养因子(brain derived neurotrophic factor antisense,BDNF-AS)的表达情况。结果与Sham组比较,I/R组大鼠海马CA1区神经元数目大幅减少,CA3和DG区神经元数目变化较小。I/R组大鼠海马CA1区中Bdnf-p1和Bdnf-p2与HDAC3结合水平明显降低(t分别为2.575和2.241,P均<0.05),Bdnf-p4和Bdnf-p6与HDAC3结合水平差异无统计学意义(t分别为1.033和0.348,P均>0.05);CA3区中Bdnf-p1和Bdnf-p2与HDAC3结合水平明显增加(t分别为12.600和3.191,P分别<0.001和<0.05),Bdnf-p6与HDAC3结合水平明显降低(t=4.029,P<0.05),Bdnf-p4与HDAC3结合水平差异无统计学意义(t=0.175,P>0.05);DG区中各BDNF启动子与HDAC3结合水平差异均无统计学意义(t=0.630~1.687,P均>0.05)。I/R组大鼠海马CA1区中BDNF-AS的表达水平明显降低(t=2.560,P<0.05),在CA3及DG区的表达水平均明显升高(t分别为3.543和3.637,P均<0.01)。结论I/R对大鼠海马中BDNF启动子与HDAC3的结合水平有显著影响,可能是通过改变BDNF-AS表达水平发挥作用。Objective To evaluate the effect of transient forebrain ischemia-reperfusion(I/R)on the binding of brainderived neurotrophic factor(BDNF)promoters to histone deacetylase 3(HDAC3)in the hippocampus of rat and investigate its mechanism.Methods The I/R model of SD rats(I/R group)was established by Pulsinelli four-vessel clamping method,and sham operation group(Sham group)was set at the same time,which were observed for the survival of neurons in the hippocampus of rats by Nissl staining,detected for the binding of BDNF promoters(Bdnf-p1,Bdnf-p2,Bdnf-p4 and Bdnf-p6)to HDAC3 by chromatin immunoprecipitation(ChIP)and determined for the expression of brain derived neurotrophic factor antisense(BDNF-AS)by qPCR.Results Compared with Sham group,the quantity of neurons in hippocampal CA1 region of rats decreased significantly in I/R group,while those in CA3 region and DG region showed no significant changes.The binding levels of Bdnf-p1 and Bdnf-p2 to HDAC3 in hippocampal CA1 region decreased significantly in I/R Group(t=2.575 and 2.241 respectively,each P<0.05),while there was no significant difference in the binding levels of Bdnf-p4 and Bdnf-p6 to HDAC3(t=1.033 and 0.348 respectively,each P>0.05);The binding levels of Bdnf-p1 and Bdnf-p2 to HDAC3 in CA3 region increased significantly(t=12.600 and 3.191,P<0.001 and<0.05,respectively),while the binding level of Bdnf-p6 to HDAC3 decreased significantly(t=4.029,P<0.05)and no significant difference was observed in the binding level of Bdnf-p4 to HDAC3(t=0.175,P>0.05);In DG region,the binding level of each BDNF promoter to HDAC3 showed no significantly difference(t=0.630~1.687,each P>0.05).Meanwhile,the expression level of BDNF-AS in hippocampal CA1 region of rats decreased significantly(t=2.560,P<0.05),but increased significantly in hippocampal CA3 and DG regions(t=3.543 and 3.637 respectively,each P<0.01)in I/R group.Conclusion I/R showed a significant effect on the binding level of BDNF promoter to HDAC3 in rat hippocampus,which may play a role by changing the expression le

关 键 词：短暂前脑缺血再灌注 脑源性神经营养因子 组蛋白去乙酰化酶 海马 

分 类 号：R338[医药卫生—人体生理学]