School of Nursing, Jiangsu Vocational College of Medicine, Yancheng, China.
Pak J Pharm Sci. 2020 May;33(3(Special)):1435-1441.
The effect of miR-532-5p on human brain microvascular endothelial cells damage induced by ox-LDL is studied. HBMEC-3 were cultured and treated with ox-LDL for 24 h to establish a model of cell oxidative damage. The expression of miR-532-5p was detected by qRT-PCR and that of CLIC4 was detected by Western blot. miR-NC, miR-532-5p mimics, si-NC, si-CLIC4, miR-532-5p mimics and pcDNA, miR-532-5p mimics and pcDNA-CLIC4 were transfected into HBMEC-3 cells, respectively, using ox -LDL processing for 24 h. Flow cytometry was used to detect the apoptotic rate. The dual luciferase reporting experiment verified the relationship between miR-532-5p and CLIC4. The ox-LDL treatment led to lower expression of miR-532-5p (p<0.05), higher expression of CLIC4 (P <0.05), enhanced content of MDA (p<0.05), decreased activities of SOD and CAT (p<0.05), increased apoptosis rate (p<0.05), higher protein level of Bax (p<0.05), and lower protein level of Bcl-2 (p<0.05). Compared with ox-LDL + miR-NC group and ox-LDL + si-NC group, ox-LDL + miR-532-5p group and ox-LDL + si-CLIC4 group had decreased content of MDA (P<0.05), increased activities of SOD and CAT (p<0.05), decreased apoptosis rate (p<0.05), lower level of Bax (p<0.05), and higher level of Bcl-2 (p<0.05). The miR-532-5p mitigates human brain microvascular endothelial cells damage induced by ox-LDL via down-regulating CLIC4 expression.
研究 miR-532-5p 对 ox-LDL 诱导的人脑血管内皮细胞损伤的影响。培养 HBMEC-3 并使用 ox-LDL 处理 24 h 建立细胞氧化损伤模型。通过 qRT-PCR 检测 miR-532-5p 的表达,通过 Western blot 检测 CLIC4 的表达。分别将 miR-NC、miR-532-5p 模拟物、si-NC、si-CLIC4、miR-532-5p 模拟物和 pcDNA、miR-532-5p 模拟物和 pcDNA-CLIC4 转染到 HBMEC-3 细胞中,用 ox-LDL 处理 24 h。流式细胞术检测细胞凋亡率。双荧光素酶报告实验验证了 miR-532-5p 与 CLIC4 的关系。ox-LDL 处理导致 miR-532-5p 表达降低(p<0.05),CLIC4 表达升高(P <0.05),MDA 含量增加(p<0.05),SOD 和 CAT 活性降低(p<0.05),细胞凋亡率增加(p<0.05),Bax 蛋白水平升高(p<0.05),Bcl-2 蛋白水平降低(p<0.05)。与 ox-LDL + miR-NC 组和 ox-LDL + si-NC 组相比,ox-LDL + miR-532-5p 组和 ox-LDL + si-CLIC4 组 MDA 含量降低(P<0.05),SOD 和 CAT 活性升高(p<0.05),细胞凋亡率降低(p<0.05),Bax 蛋白水平降低(p<0.05),Bcl-2 蛋白水平升高(p<0.05)。miR-532-5p 通过下调 CLIC4 表达减轻 ox-LDL 诱导的人脑血管内皮细胞损伤。