摘要
目的探究CXC趋化因子受体7(CXCR7)在缺血性脑卒中神经元中的功能及机制。方法采用小干扰RNA(si-RNA)技术干扰CXCR7在人神经母细胞瘤细胞SH-SY5Y细胞中的表达;在SH-SY5Y细胞中构建氧-葡萄糖剥夺/复氧(OGD/R)损伤模型;流式细胞术(FCM)检测CXCR7蛋白表达以及细胞周期;蛋白质免疫印迹试验(Western blotting)检测CXCR7和丝氨酸/苏氨酸蛋白激酶(Akt)信号通路蛋白质表达。结果与OGD/R 0 h相比,OGD/R 6 h后CXCR7表达明显降低(CXCR7/GAPDH:Western blotting检测为0.483±0.098比1.000±0.000,FCM检测为0.686±0.052比1.000±0.000,均P<0.01),细胞周期阻滞在G0/G1期(1.190±0.040比1.000±0.000,P<0.01);CXCR7 si-RNA干扰SH-SY5Y细胞后再次构建OGD/R 6 h,与相同环境下阴性对照组(si-NC组)相比,CXCR7与磷酸化Akt(p-Akt)表达明显降低(CXCR7/GAPDH:0.471±0.051比1.000±0.000,p-Akt/GAPDH:0.616±0.027比1.000±0.000,均P<0.001),并且细胞周期阻滞在G0/G1期(1.105±0.033比1.000±0.000,P<0.05)。结论CXCR7可以通过Akt信号通路调节缺血性脑卒中神经元的周期,对神经元起到保护作用。
Objective To investigate the function and mechanism of CXC chemokine receptor 7(CXCR7)in neuronal cells of ischemic stroke.Methods The expression of CXCR7 in human neuroblastoma SH-SY5Y cells was interfered by small interfering RNA(si-RNA)technique.Oxygen-glucose deprivation/reoxygenation(OGD/R)injury model was constructed in SH-SY5Y cells.CXCR7 protein expression and cell cycle were detected by flow cytometry(FCM).The protein expression of CXCR7 and Akt signaling pathway was detected by Western blotting.Results After 6 hours of OGD/R,the expression of CXCR7 was significantly decreased compared with OGD/R 0 hour(CXCR7/GAPDH:0.483±0.098 vs.1.000±0.000 by Western blotting and 0.686±0.0524 vs.1.000±0.000 by FCM,both P<0.01),cell cycle arrest in G0/G1 phase(1.190±0.040 vs.1.000±0.000,P<0.01).After CXCR7 si-RNA interference with SH-SY5Y cells,OGD/R was constructed again for 6 hours.Compared with negative control group(si-NC group)under the same environment,the expression of CXCR7 and phosphorylated Akt(p-Akt)was significantly decreased(CXCR7/GAPDH:0.471±0.051 vs.1.000±0.000,p-Akt/GAPDH:0.616±0.027 vs.1.000±0.000,both P<0.001)and cell cycle arrest in G0/G1 phase(1.105±0.033 vs.1.000±0.000,P<0.05).Conclusion The CXCR7 could regulate the cycle of neuronal cells in ischemic stroke through Akt signaling pathway,which has a protective effect on neuronal cells.
作者
王雪颖
孟海宁
王松梅
谢伟峰
曲彦
Wang Xueying;Meng Haining;Wang Songmei;Xie Weifeng;Qu Yan(Department of Medicine,Medical College of Qingdao University,Qingdao 266071,Shandong,China;Department of Critical Care Medicine,Affiliated Qingdao Municipal Hospital of Qingdao University,Qingdao 266071,Shandong,China;Department of Emergency,Affiliated Qingdao Municipal Hospital of Qingdao University,Qingdao 266071,Shandong,China)
出处
《中华危重病急救医学》
CAS
CSCD
北大核心
2022年第2期151-155,共5页
Chinese Critical Care Medicine
基金
国家自然科学基金(81971873)。
