盐酸戊乙奎醚诱导早期生长反应蛋白1的表达及其对人血管平滑肌细胞的增殖作用  

Expression of Cell Cycle Regulatory Protein 1 Induced by Penehyclidine Hydrochloride and Its Effect on Early Growth Response Protein 1

作　　者：陈曦 胡婷婷 史惠卿 刘小娟 李楠 CHEN Xi;HU Tingting;SHI Huiqing;LIU Xiaojuan;LI Nan(The General Hospitial of Western Theater Command PLA,Chengdu,Sichuan,China 610083)

机构地区：[1]中国人民解放军西部战区总医院,四川成都610083

出　　处：《中国药业》2020年第11期22-25,共4页China Pharmaceuticals

摘　　要：目的探讨早期生长反应蛋白1(EGR1)在人血管平滑肌细胞(CRL-1999)增殖过程中的作用,以及影响下游基因和盐酸戊乙奎醚对其表达的影响。方法慢病毒转染敲低CRL-1999细胞中EGR1表达,构建siEGR1-CRL-1999细胞,利用实时荧光定量聚合酶链式反应(qRT-PCR)和蛋白质印迹(Western Blot,WB)法实验验证其表达降低,同时观察下游基因细胞周期调控蛋白D1(CCND1)的表达变化。采用EDU法检测siEGR1-CRL-1999细胞的增殖能力。不同时间点观察用盐酸戊乙奎醚处理的CRL-1999细胞和siEGR1-CRL-1999细胞内EGR1蛋白的表达情况,并用WB法检测上述细胞不同时间点的磷酸化ERK1/2(p-ERK1/2)、ERK1/2蛋白表达。结果成功构建siEGR1-CRL-1999细胞;siEGR1-CRL-1999细胞增殖能力较对照组降低,且CCND1的表达相应减弱;盐酸戊乙奎醚能诱导提高CRL-1999细胞的EGR1表达水平,1 h后开始提高,2 h时达峰值,并持续至8 h时;p-ERK1/2表达在处理1 h后达峰值,并维持至2 h时,而EGR1的蛋白表达变化在时间上总落后于p-ERK1/2。结论EGR1通过CCND1影响CRL-1999细胞增殖,盐酸戊乙奎醚可能通过ERK1/2通路诱导EGR1表达。Objective To investigate the role of early growth response protein 1(EGR1)in the proliferation of human vascular smooth muscle cells(CRL-1999)and the effect of downstream genes and penehyclidine hydrochloride on its expression.Methods Lentivirus was used to knock down the expression of EGR1 in CRL-1999 cells to build siEGR1-CRL-1999 cells.The expression of EGR1 and cell cycle regulatory protein 1(CCND1)was detected by quantitative real-time polymerase chain reaction(qRT-PCR)and Western Blot(WB).The proliferation ability of siEGR1-CRL-1999 cells was examined by EDU method.CRL-1999 cells were treated with penehyclidine hydrochloride,and the expression of EGR1 p-ERK1/2 and ERK1/2 were detected at different time points.Results The siEGR1-CRL-1999 cell was successfully constructed.Compared to the control group,the proliferation of siEGR1-CRL-1999 cells was reduced,and the expression of CCND1 was also reduced.The results showed that penehyclidine hydrochloride could stimulate the expression of EGR1 in CRL-1999 cells,the EGR1 expression level began to increase at 1 h,peaked at 2 h,and continued for 8 h.After 1 h of treatment with penehyclidine phosphate,phosphorylated ERK1/2 reached the highest point of expression and remained for2 h,while the protein increase of EGR1 always lagged behind the former in time.Conclusion EGR1 affects the proliferation of CRL-1999 cells through CCND1,while penehyclidine hydrochloride can induce the expression of EGR1 through ERK1/2 pathway.

关 键 词：早期生长反应蛋白1 细胞周期调控蛋白D1 血管平滑肌细胞 戊乙奎醚 

分 类 号：R965[医药卫生—药理学]