miR-145-5p靶向FGF5对缺血/再灌注损伤诱导的神经细胞凋亡和氧化应激的调控作用
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摘要
目的研究miR-145-5p对体外缺血/再灌注(I/R)损伤神经细胞模型的调控作用,并探讨其机制。方法运用氧-葡萄糖剥夺/复氧(OGD/R)建立脑缺血/再灌注损伤细胞模型。采用荧光素酶报告实验验证miR-145-5p与FGF5的靶向关系;RT-PCR、流式细胞术、Western blot等方法检测miR-145-5p对神经细胞HT22的凋亡和氧化应激损伤的影响。结果结果表明,与未进行OGD/R处理组(Normoxia组)相比,OGD/R处理6 h、12 h、24 h组凋亡率和氧化应激损伤明显增加。在OGD/R环境下,miR-145-5p inhibitor转染减轻HT22细胞凋亡和氧化应激损伤。然而,转染FGF5 siRNA逆转了这一效应。在OGD/R模拟的脑I/R损伤过程中,miR-145-5p通过靶向FGF5促进神经细胞凋亡和损伤。结论研究结果为缺血性脑卒中的治疗提供了一种新的治疗靶点。
Objective To investigate the regulatory effects of miR-145-5 p on in vitro ischemia/reperfusion(I/R) injury of neurons and to explore their mechanisms. Methods The cell I/R injury model was established by oxygen-glucose deprivation/reoxygenation(OGD/R). The target relationship between mir-145-5 p and FGF5 was verified by luciferase reporting experiment. The effects of mir-145-5 p on apoptosis and oxidative stress in neuronal cell HT22 were detected by RT-PCR, flow cytometry and western blot. Results The results showed that the apoptosis rate and oxidative stress injury were significantly increased in the OGD/R treatment for 6 h, 12 h, and 24 h compared with the group without OGD/R treatment(Normoxia). In OGD/R model, mir-145-5 p inhibitor transfection reduced apoptosis and oxidative stress of HT22 cells. However, FGF5 siRNA transfected cells reversed this effect. In the process of brain I/R injury simulated by OGD/R, mir-145-5 p promoted apoptosis and injury of neuronal cells by targeting FGF5. Conclusion This study provides a new therapeutic target for the treatment of ischemic stroke.
Objective To investigate the regulatory effects of miR-145-5 p on in vitro ischemia/reperfusion(I/R) injury of neurons and to explore their mechanisms. Methods The cell I/R injury model was established by oxygen-glucose deprivation/reoxygenation(OGD/R). The target relationship between mir-145-5 p and FGF5 was verified by luciferase reporting experiment. The effects of mir-145-5 p on apoptosis and oxidative stress in neuronal cell HT22 were detected by RT-PCR, flow cytometry and western blot. Results The results showed that the apoptosis rate and oxidative stress injury were significantly increased in the OGD/R treatment for 6 h, 12 h, and 24 h compared with the group without OGD/R treatment(Normoxia). In OGD/R model, mir-145-5 p inhibitor transfection reduced apoptosis and oxidative stress of HT22 cells. However, FGF5 siRNA transfected cells reversed this effect. In the process of brain I/R injury simulated by OGD/R, mir-145-5 p promoted apoptosis and injury of neuronal cells by targeting FGF5. Conclusion This study provides a new therapeutic target for the treatment of ischemic stroke.
引文
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[2] Zhai W W,Sun L,Yu Z Q,et al.Hyperbaric oxygen therapy in experimental and clinical stroke[J].Med Gas Res,2016,6(2):111-8.
[3] Fluri F,Schuhmann M K,Kleinschnitz C.Animal models of ischemic stroke and their application in clinical research[J].Drug Des Devel Ther,2015,9:3445-54.
[4] Di Y,Lei Y,Yu F,et al.Micrornas expression and function in cerebral ischemia reperfusion injury[J].J Mol Neurosci,2014,53(2):242-50.
[5] Qi X,Shao M,Sun H,et al.Long non-coding rna snhg14 promotes microglia activation by regulating mir-145-5p/pla2g4a in cerebral infarction[J].Neuroscience,2017,348:98-106.
[6] Xu B,Ji X,Chen X,et al.Effect of perfluorooctane sulfonate on pluripotency and differentiation factors in mouse embryoid bodies[J].Toxicology,2015,328:160-7.
[7] Donnan G A,Fisher M,Macleod M,et al.Stroke[J].Lancet (London,England),2008,371(9624):1612-23.
[8] Lipton P.Ischemic cell death in brain neurons[J].Physiol Rev,1999,79(4):1431-568.
[9] Chen Y,Wang X,Yang M,et al.Mir-145-5p increases osteoclast numbers in vitro and aggravates bone erosion in collagen-induced arthritis by targeting osteoprotegerin[J].Med Sci Monit,2018,24:5292-300.
[10] Cui L,Fang L,Mao X,et al.Gdnf-induced downregulation of mir-145-5p enhances human oocyte maturation and cumulus cell viability[J].J Clin Endocrinol Metab,2018,103(7):2510-21.
[11] Chang Y,Yan W,Sun C,et al.Mir-145-5p inhibits epithelial-mesenchymal transition via the jnk signaling pathway by targeting map3k1 in non-small cell lung cancer cells[J].Oncol Lett,2017,14(6):6923-8.
[12] Kehler J S,David V A,Schaffer A A,et al.Four independent mutations in the feline fibroblast growth factor 5 gene determine the long-haired phenotype in domestic cats[J].J Hered,2007,98(6):555-66.
[13] Fang F,Chang R M,Yu L,et al.Microrna-188-5p suppresses tumor cell proliferation and metastasis by directly targeting fgf5 in hepatocellular carcinoma[J].J Hepatol,2015,63(4):874-85.
[14] Hanaka H,Hamada T,Ito M,et al.Fibroblast growth factor-5 participates in the progression of hepatic fibrosis[J].Exp Anim,2014,63(1):85-92.
[15] Park G C,Song J S,Park H Y,et al.Role of fibroblast growth factor-5 on the proliferation of human tonsil-derived mesenchymal stem cells[J].Stem Cells Dev,2016,25(15):1149-60.