摘要
该文旨在探讨组蛋白去乙酰化酶6(histone deacetytase 6,HDAC6)通过调控热休克蛋白90(heat shock protein 90,HSP90)影响体外培养Aβ诱导的大鼠海马神经元细胞功能及形态的变化。取孕18天SD大鼠胎鼠,体外培养海马神经元细胞,7天后用β-tubulin III抗体鉴定海马神经元纯度。用寡聚体Aβ1-42干预24 h,随后分别加入HDAC6抑制剂TSA、HDAC6激动剂Theo、HSP90抑制剂Gane或生理盐水;细胞不加Aβ1-42干预作为对照。用CCK8法检测细胞的活力,免疫荧光法观察神经元细胞突起的长度及形态变化,Western blot检测HDAC6和HSP90蛋白表达,qRT-PCR检测hsp90基因的mRNA表达水平。结果表明,应用HDAC6抑制剂可下调HDAC6水平,同时促进了hsp90 mRNA和HSP90蛋白的表达,也提高了海马神经元活性、突起长度和分支数目,HDAC6激动剂则引起相反的效应;而应用HSP90抑制剂后则降低了神经元活性和突起长度以及分支数目,但HDAC6没有变化。因此,推测HDAC6可能通过调控HSP90水平影响Aβ诱导的大鼠海马神经元细胞功能和形态的变化。
The aim of this study was to investigate the effect of HDAC6(histone deacetyltase 6)on the functional and morphology of the Aβ(amyloid beta)-induced primary hippocampal neurons by regulating HSP90.In vitro primary culture was conducted on the hippocampal neurons of 18 days pregnant SD rats,the purity of hippocampal neurons was identified withβ-tubulin III antibody after 7 days.After oligomeric-Aβ1-42(oligomeric-amyloid beta peptides 1-42)cultured 24 hours,a variety of drugs including HDAC6 inhibitor TSA(tubastatin A hydrochbride),HDAC6 agonist Theo(theophylline),HSP90 inhibitor Gane(ganetespi),or saline(Aβ+NS group)were added and then cultured 24 h,Saline group as a control(NS group).Then cell viability was detected by CCK-8 assay,the length and branchs of neurons were used with immunofluorescence staining,the levels of HDAC6 and HSP90 protein were detected by Western blot,and the mRNA expressions of hsp90 were performed to use qRT-PCR(quantitative Real-time polymerase chain reaction).The results showed that HDAC6 hibitor could not only down-regulate the level of HDAC6 and promote the expressions of hsp90 mRNA and HSP90 protein,but also could increase the activity of hippocampal neurons,the length of protuberances and the number of branches.However,there appeared to be an opposite effect in the agonist of HDAC6.Moreover,the cell viability,synaptic length and number of branches after the application of HSP90 inhibitor were decreased,but no significant changes in the level of HDAC6.Thus,we speculate that HDAC6 may influence the functional and morphology of the Aβ-induced primary hippocampal neurons by regulating HSP90 levels.
作者
刘镇
郝凯敏
祁文秀
LIU Zhen;HAO Kaimin;QI Wenxiu(Fenyang College of Shanxi Medical University,Fenyang 032200,China;Department of Physiology,Fenyang College of Shanxi Medical University,Fenyang 032200,China)
出处
《中国细胞生物学学报》
CAS
CSCD
2020年第4期549-556,共8页
Chinese Journal of Cell Biology
基金
山西省自然科学基金(批准号:201801D121348)
山西医科大学汾阳学院科研项目(批准号:2018C06)资助的课题。
