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EGb761 Protects Brain Microvascular Endothelial Cells Against Oxygen–Glucose Deprivation-Induced Injury Through lncRNA Rmst/miR-150 Axis

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Abstract

In the present study, we aimed to illustrate the roles and working mechanisms of long non-coding RNA (lncRNA) rhabdomyosarcoma 2-associated transcript (Rmst) and EGb761 in oxygen–glucose deprivation (OGD)-induced brain microvascular endothelial cells (BMECs). OGD exposure augmented the level of Rmst while reduced the expression of miR-150 in bEnd.3 cells. MiR-150 could directly bind to Rmst in bEnd.3 cells. Rmst silencing abrogated the inhibitory influences on the proliferation and migration and the promoting impact on the apoptosis of bEnd.3 cells caused by OGD exposure. Rmst overexpression intensified OGD-induced injury in bEnd.3 cells. OGD induced the injury of bEnd.3 cells through Rmst/miR-150 axis. EGb761 attenuated the damage in bEnd.3 cells induced by OGD through targeting Rmst/miR-150 axis. EGb761 might be an effective therapeutic agent to protect brain microvascular endothelial cells from hypoxia–ischemia induced injury.

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Abbreviations

Rmst:

Rhabdomyosarcoma 2-associated transcript

RIP:

RNA immune co-precipitation

OGD:

Oxygen–glucose deprivation

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Authors and Affiliations

  1. Department of Traditional Chinese Medicine, Yantaishan Hospital of Yantai, Yantai, Shandong, China

    Peng Qiao

  2. Department of Clinical Laboratory, Shandong Laiyang Health School, Laiyang, Shandong, China

    Huijing Yan

  3. College of Traditional Chinese Medicine, Shandong University of Traditional Chinese Medicine, No. 4655 Daxue Road, Changqing District, Jinan, 250355, Shandong, China

    Jin Wang

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Correspondence to Jin Wang.

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11064_2020_3099_MOESM1_ESM.tif

Supplementary Figure 1. Rmst silencing has no significant influence on the proliferation, migration and apoptosis of bEnd.3 cells without OGD exposure. (A-G) bEnd.3 cells were transfected with si-con or si-Rmst. (A) The level of Rmst in transfected bEnd.3 cells was examined by qRT-PCR assay. Si-Rmst group/si-con group: 0.0003. (B) Cell proliferation was analyzed via MTT assay. (C) Scratch wound assay was used to analyze the migration ability of bEnd.3 cells. (D and E) Western blot assay was conducted to measure the protein levels of MMP2 and MMP9 in bEnd.3 cells. (F) Flow cytometry was used to determine the apoptotic rate of bEnd.3 cells. (G) Caspase 3 Activity Assay Kit was utilized to measure the activity of caspase-3 in bEnd.3 cells. *P<0.05. Supplementary file1 (TIF 1553 kb)

11064_2020_3099_MOESM2_ESM.tif

Supplementary Figure 2. EGb761 down-regulates Rmst level while enhances the level of miR-150 in bEnd.3 cells. (A and B) The abundance of Rmst and miR-150 in bEnd.3 cells treated with or without EGb761 was examined by qRT-PCR. Rmst, EGb761 group/Control group: 0.0314. MiR-150, EGb761 group/Control group: 0.0325. *P<0.05. Supplementary file2 (TIF 433 kb)

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Qiao, P., Yan, H. & Wang, J. EGb761 Protects Brain Microvascular Endothelial Cells Against Oxygen–Glucose Deprivation-Induced Injury Through lncRNA Rmst/miR-150 Axis. Neurochem Res 45, 2398–2408 (2020). https://doi.org/10.1007/s11064-020-03099-8

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