FABP4对高糖诱导的ARPE-19细胞凋亡及炎症效应的影响  

Effects of FABP4 on Apoptosis and Inflammation of ARPE-19 Cells Induced by High Glucose

作　　者：原霞 王越 任欣会 YUAN Xia;WANG Yue;REN Xinhui(Department of Endocrinology,Zhengzhou Yi He Hospital,Zhengzhou Henan 450000,China)

机构地区：[1]郑州颐和医院内分泌科,河南郑州450000

出　　处：《联勤军事医学》2023年第3期195-200,共6页Military Medicine of Joint Logistics

摘　　要：目的探讨脂肪酸结合蛋白4(fatty acid binding protein 4,FABP4)在高糖诱导的人视网膜色素上皮细胞株(adult retinal pigment epithelial cell line 19,ARPE-19)凋亡及炎症效应中的作用。方法根据培养条件,将ARPE-19细胞分成Control组(5.5 mmol/L葡萄糖)、OS组(5.5 mmol/L葡萄糖+19.5 mmol/L甘露醇)和HG组(25 mmol/L葡萄糖)。各组ARPE-19细胞培养24 h后,采用实时荧光逆转录定量聚合酶链反应(real-time reverse transcription quantitative polymerase chain reaction,RT-qPCR)及Western blot检测FABP4的表达。si-FABP4转染ARPE-19细胞后,构建高糖刺激模型,Annexin V-异硫氰酸荧光素(fluorescein isothiocyanate,FITC)/碘化丙啶(propidium iodide,PI)双染法检测细胞凋亡,RT-qPCR及酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)分别检测肿瘤坏死因子α(tumor necrosis factor alpha,TNF-α)、白细胞介素6(interleukin 6,IL-6)、IL-1β在细胞及培养基中的表达水平,Western blot检测凋亡相关蛋白[Bad、B淋巴细胞瘤2基因(B-cell lymphoma 2,Bcl-2)、Cleaved caspase-3]表达及核因子κB(nuclear factor kappa B,NF-κB)通路激活。结果与Control组比较,HG中FABP4 mRNA及FABP4蛋白的表达均显著升高(P均<0.01)。与HG组比较,si-FABP4抑制后,细胞凋亡率显著降低(P<0.01),Bad、Cleaved caspase-3蛋白表达被抑制(P均<0.01),而Bcl-2的表达显著增加(P<0.05);ARPE-19细胞内和ARPE-19细胞培养基中炎症因子(TNF-α、IL-6、IL-1β)的表达量均显著降低(P均<0.01),同时ARPE-19细胞中p-p65、p-IκBα的表达被显著抑制(P均<0.01)。结论FABP4参与高糖诱导的ARPE-19细胞凋亡及炎症释放,其机制可能与NF-κB通路激活有关。Objective To investigate the role of fatty acid binding protein 4(FABP4)on the apoptosis and inflammatory of adult retinal pigment epithelial cell line 19(ARPE-19)induced by high glucose.Methods ARPE-19 cells were divided into Control group(5.5 mmol/L glucose),OS group(5.5 mmol/L glucose+19.5 mmol/L mannitol)and HG group(25 mmol/L glucose).ARPE-19 cells in each group were cultured for 24 hours,after that FABP4 expression was detected by real-time reverse transcription quantitative polymerase chain reaction(RT-qPCR)and Western blot.si-FABP4 was transfected into ARPE-19 cells to construct a high glucose stimulation model,cell apoptosis was detected by Annexin V-fluorescein isothiocyanate(FITC)/propidium iodide(PI)double staining,and the expression levels of tumor necrosis factor alpha(TNF-α),interleukin 6(IL-6)and IL-1βin cells and culture medium were detected by RT-qPCR and enzyme-linked immunosorbent assay(ELISA)respectively,the expression of apoptosis-related proteins[Bad,B-cell lymphoma 2(Bcl-2),Cleaved caspase-3]and activation of nuclear factor kappa B(NF-κB)pathway were detected by Western blot.Results Compared with the Control group,the expression of FABP4 mRNA and FABP4 protein in HG group significantly increased(all P<0.01).Compared with HG group,the apoptosis rate significantly decreased after si-FABP4 inhibition(P<0.01),the expression of Bad and Cleaved caspase-3 protein was inhibited(all P<0.01),while the expression of Bcl-2 significantly increased(P<0.05);the expression of inflammatory factors(TNF-α,IL-6,IL-1β)in ARPE-19 cells and in the culture medium of ARPE-19 cells significantly decreased(all P<0.01),in the meantime,the expressions of p-p65 and p-IκBαin ARPE-19 cells were significantly inhibited(all P<0.01).Conclusion FABP4 is involved in the apoptosis and inflammatory release of ARPE-19 induced by high glucose,and the mechanism may be related to the activation of NF-κB pathway.

关 键 词：脂肪酸结合蛋白4 糖尿病视网膜病变 炎症释放 细胞凋亡 核因子ΚB 

分 类 号：R587.2[医药卫生—内分泌]