The aims of this study were to find a treatment for acute kidney injury in sepsis and study the role of miR-128-3p in this process. We generated a model of septic acute kidney injury through cecal ligation and puncture (CLP) induction and screened differentially expressed microRNAs through microarray. The mechanism used by miR-128-3p in inflammatory response to septic acute kidney injury was investigated using cell transfection assay, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), flow cytometry, enzyme-linked immunosorbent assay, western blot, and Dual-Luciferase Reporter Assay. miRNA microarray screening revealed that miR-128-3p was significantly upregulated in the kidneys of mice with CLP-induced septic acute kidney injury. The level of inflammatory factors TNF-α, IL-1 β, and IL-6 decreased. In contrast, cell viability increased and apoptosis decreased with the addition of miR-128-3p inhibitors in TCMK-1 cells treated with lipopolysaccharide (LPS). Using bioinformatics and luciferin reporter gene experiments, we found that NRP1 is a miR-128-3p target gene. Overexpression of NRP1 in LPS-treated TCMK-1 cells decreased the expression of TNF-α, IL-6, and IL-1β; increased cell viability; and decreased apoptosis. The survival period of mice pretreated with miR-128-3p inhibitors was prolonged, infiltration of inflammatory cells into kidney tissue decreased, permeability of kidneys enhanced, and expression of inflammatory factors and renal apoptosis decreased. miR-128-3p targets NRP1 for cell degradation, promotes inflammatory cell infiltration, increases expression of inflammatory factors, decreases renal cell viability, and increases apoptosis in LPS-induced septic acute renal injury.

这项研究的目的是寻找一种脓毒症急性肾脏损伤的治疗方法，并研究miR-128-3p在此过程中的作用。我们通过盲肠结扎和穿刺（CLP）诱导生成了败血性急性肾损伤模型，并通过微阵列筛选了差异表达的microRNA。使用细胞转染测定，3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四唑溴化物（MTT），流式细胞仪研究了miR-128-3p对败血性急性肾损伤的炎症反应所使用的机制，酶联免疫吸附测定，蛋白质印迹和Dual-Luciferase Reporter Assay。miRNA基因芯片筛选显示，miR-128-3p在患有CLP诱导的败血性急性肾损伤的小鼠肾脏中显着上调。炎性因子TNF-α，IL-1β和IL-6的水平下降。相反，在脂多糖（LPS）处理的TCMK-1细胞中，添加miR-128-3p抑制剂可增强细胞活力，减少凋亡。使用生物信息学和荧光素报告基因实验，我们发现NRP1是miR-128-3p靶基因。LRP处理的TCMK-1细胞中NRP1的过表达降低了TNF-α，IL-6和IL-1β的表达。细胞活力增加；并减少凋亡。经miR-128-3p抑制剂预处理的小鼠的生存期延长，炎症细胞向肾脏组织的浸润减少，肾脏的通透性增强，炎症因子的表达和肾细胞凋亡减少。miR-128-3p靶向NRP1进行细胞降解，促进炎性细胞浸润，增加炎性因子的表达，降低肾细胞活力，