1 Memory T Cell Laboratory, Department of Immunology, School of Medicine, Shiraz University of Medical Science, Shiraz, Iran.
2 Department of Obstetrics and Gynecology, School of Medicine, Shiraz University of Medical Science, Shiraz, Iran.
Int J Toxicol. 2017 Mar/Apr;36(2):133-141. doi: 10.1177/1091581817695270.
Inflammation and oxidative stress are important risk factors affecting various cells in the formation of atherosclerosis. MicroRNAs (miRs) are regulators of inflammation and atherogenesis. The expressions of endothelial cell (EC)-specific miR-10a and miR-21 and monocyte-specific miR-33a and miR-221 were investigated using coculture of the ECs and monocytes upon exposure to HO as an oxidative stressor, and endotoxin/lipopolysaccharide (LPS) as a microbial stressor. Human umbilical endothelial cells (HUVECs) and peripheral blood mononuclear cells (or monocytes) were cocultured in M199 complete medium and were incubated with LPS (20 ng/mL) or HO (1%) for 8 hours at 37°C. The HUVECs and monocytes were then separated from the cellular mix using a magnetic bead negative selection technique. The relative expression of miRs was determined by real-time polymerase chain reaction. In both cell types, HO induced miR10a ( P = 0.05) and LPS induced miR21 ( P = 0.0003) compared to the untreated controls. Coculture increased miR-10a and miR-21 expression in monocytes ( P = 0.0008 and <0.0001); however when cultured alone, HUVECs expressed higher levels of miR-10a and miR-21 ( P < 0.0001 and <0.0001). Coculture decreased the expression of miR-33a in monocytes ( P < 0.0001) while increasing miR221 in HUVECs and monocytes ( P < 0.0001 and <0.0001). The expression pattern of miRs in HUVECs and monocytes changes in the coculture compared to culturing alone in response to oxidative and microbial toxic compounds. Moreover, different cellular stressors induce different athero-miRs, which may affect the course of inflammation.
炎症和氧化应激是影响动脉粥样硬化形成的各种细胞的重要危险因素。微小 RNA（miRs）是炎症和动脉粥样硬化形成的调节因子。通过将内皮细胞（EC）特异性 miR-10a 和 miR-21 以及单核细胞特异性 miR-33a 和 miR-221 的表达与暴露于 HO 作为氧化应激物以及内毒素/脂多糖（LPS）作为微生物应激物的 EC 和单核细胞共培养来研究。将人脐静脉内皮细胞（HUVEC）和外周血单核细胞（或单核细胞）共培养在 M199 完全培养基中，并在 37°C 下用 LPS（20ng/mL）或 HO（1%）孵育 8 小时。然后使用磁珠阴性选择技术将 HUVEC 和单核细胞从细胞混合物中分离出来。通过实时聚合酶链反应确定 miRs 的相对表达。在两种细胞类型中，与未处理的对照相比，HO 诱导 miR10a（P=0.05）和 LPS 诱导 miR21（P=0.0003）。共培养增加了单核细胞中 miR-10a 和 miR-21 的表达（P=0.0008 和 <0.0001）；然而，当单独培养时，HUVEC 表达更高水平的 miR-10a 和 miR-21（P<0.0001 和 <0.0001）。共培养降低了单核细胞中 miR-33a 的表达（P<0.0001），同时增加了 HUVEC 和单核细胞中 miR221 的表达（P<0.0001 和 <0.0001）。与单独培养相比，miRs 在 HUVEC 和单核细胞中的表达模式在共培养中发生变化，以响应氧化和微生物毒性化合物。此外，不同的细胞应激源诱导不同的动脉粥样硬化 miR，这可能影响炎症过程。
