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缺氧诱导因子-1α 的过表达通过降低 miR-134 的表达来保护 PC12 细胞免受 OGD/R 诱导的损伤。

Overexpression of HIF-1α protects PC12 cells against OGD/R-evoked injury by reducing miR-134 expression.

机构信息

Department of Neurosurgery, The Affiliated Hospital of Qingdao University, Qingdao, Shandong, China.

Department of Neurosurgery, Qingdao Municipal Hospital, Qingdao, Shandong, China.

出版信息

Cell Cycle. 2020 May;19(9):990-999. doi: 10.1080/15384101.2020.1743903. Epub 2020 Apr 8.

Abstract

Nowadays, searching for new therapeutic targets for cerebral stroke treatment are still in urgent need. Our study explored the influences and mechanisms of HIF-1α on OGD/R-evoked injury. OGD/R treatment was conducted on PC12 cells to simulate ischemic injury. CCK-8, flow cytometry and qRT-PCR were conducted to determine the variations of cell viability, apoptosis and gene expression, respectively. Cell transfections were conducted to overexpress HIF-1α and miR-134. Variations of protein levels were evaluated by employing western blot. Results showed that OGD/R treatment induced cell injury through reducing viability, while enhancing apoptosis that was validated by the elevated ratios of C/P-PARP and C/P-caspase-3. HIF-1α expression was markedly increased by OGD/R treatment. HIF-1α overexpression attenuated OGD/R-evoked injury in PC12 cells and remarkably reversed OGD/R-triggered inhibitory effects on ERK1/2 and JAK1/STAT3 pathways. Besides, miR-134 was also down-regulated by HIF-1α overexpression in PC12 cells. Up-regulation of miR-134 notably counteracted HIF-1α overexpression-triggered neuro-protective impacts on OGD/R-evoked injury and ERK1/2 and JAK1/STAT3 pathways. Our present study reported that HIF-1α overexpression protected PC12 cells against OGD/R-evoked injury via down-regulation of miR-134, which making HIF-1α and miR-134 to be promising targets for cerebral stroke therapy.

摘要

如今,寻找治疗脑卒中新的治疗靶点仍然迫在眉睫。我们的研究探讨了 HIF-1α 对 OGD/R 诱导损伤的影响及其机制。OGD/R 处理在 PC12 细胞上进行,以模拟缺血性损伤。CCK-8、流式细胞术和 qRT-PCR 分别用于确定细胞活力、凋亡和基因表达的变化。细胞转染用于过表达 HIF-1α 和 miR-134。通过 Western blot 评估蛋白水平的变化。结果表明,OGD/R 处理通过降低细胞活力诱导细胞损伤,同时通过增加 C/P-PARP 和 C/P-caspase-3 的比值来增强凋亡。OGD/R 处理显著增加了 HIF-1α 的表达。HIF-1α 的过表达减轻了 PC12 细胞中的 OGD/R 诱导损伤,并显著逆转了 OGD/R 对 ERK1/2 和 JAK1/STAT3 途径的抑制作用。此外,miR-134 也被 HIF-1α 过表达下调。miR-134 的上调显著抵消了 HIF-1α 过表达对 OGD/R 诱导损伤和 ERK1/2 和 JAK1/STAT3 途径的神经保护作用。本研究报道,HIF-1α 的过表达通过下调 miR-134 来保护 PC12 细胞免受 OGD/R 诱导的损伤,这使得 HIF-1α 和 miR-134 成为治疗脑卒中的有前途的靶点。

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