Yang Xiao-Lan, Mi Jian-Hua, Dong Qing
Department of Neurology, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 201100, P.R. China.
Exp Ther Med. 2021 Mar;21(3):181. doi: 10.3892/etm.2021.9612. Epub 2021 Jan 5.
Ischemic stroke is a life-threatening complication with a high rate of morbidity. Circulating fatty acid binding protein 4 (FABP4) has been reported to be associated with the outcome of acute ischemic stroke. The present study aimed to illustrate the function of FABP4 in ischemic stroke. PC12 cells exposed to oxygen glucose deprivation/reoxygenation (OGD/R) were used to mimic ischemia-reperfusion (I/R) injury in ischemic stroke. Cell viability was estimated using a Cell Counting Kit-8 assay. The expression of FABP4 in PC12 cells under OGD/R was detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). PC12 cells were transfected with FABP4 small interfering RNA (siRNA), inflammatory cytokines and reactive oxygen species (ROS) were determined via RT-qPCR and ROS assay kit. Western blotting was performed to detect endoplasmic reticulum stress (ERS)-related proteins and peroxisome proliferator-activated receptor γ (PPARγ). Flow cytometry was used to evaluate the cell apoptotic rate. The expression of FABP4 increased gradually with the prolongation of reoxygenation within 8 h. FABP4-knockdown inhibited the transcription of inflammatory cytokines, the production of ROS and decreased cell apoptosis. Furthermore, decreased ERS-related proteins and increased PPARγ were estimated in PC12 cells transfected with FABP4 siRNA. PPARγ inhibitor GW9662 weakened the anti-apoptotic effect of FABP4-knockdown. Taken together, these results indicated that FABP4-knockdown suppressed cell apoptosis via relieving ERS; this effect was reversed by treatment of GW9662.
缺血性中风是一种危及生命的并发症，发病率很高。据报道，循环脂肪酸结合蛋白4（FABP4）与急性缺血性中风的预后相关。本研究旨在阐明FABP4在缺血性中风中的作用。将暴露于氧糖剥夺/复氧（OGD/R）的PC12细胞用于模拟缺血性中风中的缺血再灌注（I/R）损伤。使用细胞计数试剂盒-8检测法评估细胞活力。通过逆转录定量聚合酶链反应（RT-qPCR）检测OGD/R条件下PC12细胞中FABP4的表达。用FABP4小干扰RNA（siRNA）转染PC12细胞，通过RT-qPCR和ROS检测试剂盒测定炎性细胞因子和活性氧（ROS）。进行蛋白质免疫印迹法检测内质网应激（ERS）相关蛋白和过氧化物酶体增殖物激活受体γ（PPARγ）。采用流式细胞术评估细胞凋亡率。在8小时内，随着复氧时间的延长，FABP4的表达逐渐增加。敲低FABP4可抑制炎性细胞因子的转录、ROS的产生并减少细胞凋亡。此外，在转染FABP4 siRNA的PC12细胞中，ERS相关蛋白减少，PPARγ增加。PPARγ抑制剂GW9662减弱了敲低FABP4的抗凋亡作用。综上所述，这些结果表明，敲低FABP4通过减轻ERS抑制细胞凋亡；GW9662处理可逆转这种作用。
