核受体NR6A1通过上调RIPK3基因表达诱导血管平滑肌细胞凋亡

发布时间：2018-05-29 04:02

本文选题：核受体亚家族A + 受体相互作用丝氨酸/苏氨酸蛋白激酶　；参考：《中国病理生理杂志》2017年09期

【摘要】：目的:探讨核受体亚家族6A1(NR6A1)对血管平滑肌细胞凋亡的影响及可能的分子机制。方法:将腺病毒Ad-NR6A1感染大鼠血管平滑肌细胞,分别在感染后0 h、24 h和48 h时进行MTT实验,以时间为横坐标,A570为纵坐标绘制细胞生长曲线,观察NR6A1对细胞生长的影响;进行DAPI染色、TUNEL染色及caspase活性检测,观察细胞凋亡情况;进一步通过基因芯片技术,寻找NR6A1的靶基因;采用siRNA介导的基因沉默技术,观察受体相互作用丝氨酸/苏氨酸蛋白激酶3(RIPK3)基因沉默对NR6A1诱导的血管平滑肌细胞凋亡的影响。结果:腺病毒Ad-NR6A1感染细胞48 h时,NR6A1过表达组细胞数量较对照组(重组腺病毒载体Ad-Lac Z)明显减少;DAPI染色显示NR6A1过表达诱导血管平滑肌细胞出现核浓缩和核碎裂的凋亡表型,TUNEL染色显示NR6A1过表达引起细胞凋亡,caspase活性检测结果显示NR6A1过表达细胞内caspase-3、caspase-8和caspase-9活性均较对照组高;基因芯片技术检测发现,NR6A1过表达上调血管平滑肌细胞中RIPK3基因表达;RIPK3基因沉默可以显著抑制NR6A1诱导的平滑肌细胞凋亡。结论:NR6A1通过上调RIPK3基因表达诱导血管平滑肌细胞凋亡。
[Abstract]:Aim: to investigate the effect of nuclear receptor subfamily 6A1NR6A1 on apoptosis of vascular smooth muscle cells (VSMCs) and its possible molecular mechanism. Methods: rat vascular smooth muscle cells (VSMCs) were infected with adenovirus Ad-NR6A1. The MTT experiments were carried out at 24 h and 48 h after infection respectively. The cell growth curves were drawn with the time axis A570 as the vertical coordinate to observe the effect of NR6A1 on the cell growth. DAPI staining, Tunel staining and caspase activity detection were used to observe the apoptosis of cells; to further search for the target genes of NR6A1 by gene chip technology; to use siRNA mediated gene silencing, To observe the effect of receptor interaction serine / threonine protein kinase 3 (RIPK3) gene silencing on apoptosis of vascular smooth muscle cells induced by NR6A1. Results: after 48 h of adenovirus Ad-NR6A1 infection, the number of NR6A1 overexpression group was significantly lower than that of control group (recombinant adenovirus vector Ad-Lac Z). NR6A1 overexpression induced nuclear condensation and nuclear fragmentation in vascular smooth muscle cells. Apoptosis induced by NR6A1 overexpression was detected by Tunel staining. The results showed that the activities of caspase-3, caspase-8 and caspase-9 in NR6A1 overexpression cells were higher than those in control group. Gene chip technique showed that the overexpression of NR6A1 up-regulated the expression of RIPK3 gene and the silencing of RIPK3 gene could significantly inhibit the apoptosis of smooth muscle cells induced by NR6A1. Conclusion the apoptosis of vascular smooth muscle cells was induced by up-regulating the expression of RIPK3 gene in vascular smooth muscle cells.
【作者单位】：湖北医药学院病理生理学教研室;
【基金】：湖北省教育厅科学研究计划指导性项目(No.B2015492) 湖北省自然科学基金资助项目(No.2014CFB187) 国家自然科学基金资助项目(No.81641140)
【分类号】：R363

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