微小RNA-138/NGAL对缺血再灌注诱导人肾小管上皮细胞的影响

目的 探究微小RNA-138(microRNA-138,miR-138)调控中性粒细胞明胶酶相关脂质运载蛋白(neutrophil gelatinase-associated lipocalin,NGAL)对缺血/再灌注(ischemia/reperfusion,I/R)诱导人肾小管上皮(HK-2)细胞损伤的影响。方法 HK-2细胞构建I/R模型,分别转染miR-138模拟物、miR-138抑制剂、NGAL、NGAL+miR-138模拟质粒,qRT-PCR测定不同细胞miR-138或NGAL mRNA表达鉴定转染结果,细胞活力检测(cell counting kit-8,CCK-8)法和流式细胞术测定细胞活性和凋亡。ELISA、Western blot法分别测定miR-138模拟物、miR-138抑制剂对I/R细胞白细胞介素(interleukin,IL)-6、IL-1β、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平,以及Toll样受体4(toll like receptor 4,TLR4)、核因子κB(nuclear factor kappa-B...

Effect of microRNA-138 on the ischemia/reperfusion of human renal tubular epithelial cells

Objective To explore the effect of microRNA-138 (miR-138) on injury of ischemia/reperfusion (I/R) induced human renal tubular epithelium (HK-2) cells through neutrophil gelatinase-associated lipocalin (NGAL). Methods HK-2 cells were used to construct I/R model cells, and transfected with miR-138 mimic, miR-138 inhibitor, NGAL, NGAL + miR-138 mimic plasmids, respectively. qRT-PCR determined the expression of miR-138 or NGAL mRNA in different cells to identify the transfection results. Cell counting kit-8 (CCK-8) method and flow cytometry were used to detected the activities and apoptosis of cells. ELISA and western blot were used to determine the effects of miR-138 mimic or miR-138 inhibitor on levels of interleukin (IL)-6, IL-1β, tumor necrosis factor (TNF-α) and protein expression of toll like receptor 4 (TLR4), nuclear factor kappa-B (NF-κB), inhibitor of NF- κB (IκBα), pho-IκBα (p-IκBα), NGAL of cells. Results miR-138 mRNA expression and cell activity were decreased, while apoptosis increased in I/R cells (P<0.01). Plasmid transfected well, miR-138 mimic increased activity while decreased apoptosis and NGAL mRNA expression of I/R cell. miR-138 inhibitor or NGAL mimic inhibited activity and increased apoptosis and NGAL mRNA expression of I/R cell. The negative effects of NGAL mimic on I/R cell were reversed by miR-138 mimic. miR-138 inhibitor increased levels of IL-6, IL-1β, TNF-α of I/R cell, and increased TLR4, NF-κB, p-IκBα, NGAL protein expression and decreased IκBα protein expression (P<0.05). While miR-138 mimic decreased levels of IL-6, IL-1β, TNF-α of I/R cell, and decreased TLR4, NF-κB, p-IκBα, NGAL protein expression and increased IκBα protein expression (P<0.05). Conclusion miR-138 reduced apoptosis and inflammation factor levels to play a protective role on I/R induced HK-2 cells may through regulating NGAL and TLR4/NF- κB pathway.