Department of Nephrology, Qingdao Municipal Hospital, Qingdao, China.
Eur Rev Med Pharmacol Sci. 2019 Aug;23(16):7024-7032. doi: 10.26355/eurrev_201908_18744.
To explore the effect of micro ribonucleic acid (miRNA)-146a on kidney injury in mice with systemic lupus erythematosus (SLE), and to investigate its possible mechanism.
A total of 45 female MRL/lpr mice were randomly divided into control group, miR-146a mimic group and miR-146a inhibitor group. Urine protein level was measured every 2 weeks. Meanwhile, the levels of serum anti-dsdeoxyribonucleic acid (anti-dsDNA), anti-ssDNA, antinuclear antibody (ANA) and anti-chromatin were measured using enzyme-linked immunosorbent assay (ELISA). At 2 weeks after drug treatment, the effects of miR-146a mimic and inhibitor on kidney tissues of MRL/lpr mice were detected and analyzed by gene chip and gene set enrichment analysis, respectively. The mice were executed at the age of 24 weeks, and the blood samples were collected. Subsequently, the level of blood urea nitrogen (BUN) was measured using the BUN analyzer. After that, kidney tissues were taken, and the effect of drug treatment on the morphology of kidney tissues was detected via hematoxylin-eosin (HE) staining. Moreover, the effects of drug treatment on the mRNA levels of inflammatory factors and the nuclear factor-κB (NF-κB) signaling pathway in kidney tissues were detected via quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting, respectively.
MiR-146a mimic significantly reduced urine protein in a time-dependent manner, which also significantly reduced BUN level at 24 weeks. The results of HE staining showed that both glomerular injury and renal vascular injury in miR-146a mimic group were significantly alleviated. In miR-146a mimic group, serum autoantibodies of anti-dsDNA, anti-ssDNA, anti-chromatin and ANA decreased significantly. However, the survival time of mice was significantly prolonged. High-throughput gene expression chip technique elucidated that in miR-146a mimic group, the expression of positive regulatory gene of NF-κB showed a decreasing trend. However, the expression of negative regulatory gene of NF-κB showed an increasing trend. MiR-146a mimic remarkably down-regulated the expression levels of RELA, IRAK1, interleukin-1B (IL1B) and IL-10 in kidney tissues. Furthermore, the results of Western blotting showed that miR-146a mimic inhibited both the classical and non-classical NF-κB signaling pathways.
MiR-146a reduces SLE-induced kidney injury in MRL/lpr mice through regulating classical and non-classical NF-κB signaling pathways.
探讨微小 RNA-146a（miRNA-146a）对系统性红斑狼疮（SLE）小鼠肾损伤的影响，并探讨其可能的机制。
将 45 只雌性 MRL/lpr 小鼠随机分为对照组、miR-146a 模拟物组和 miR-146a 抑制剂组。每两周测量一次尿蛋白水平。同时，采用酶联免疫吸附试验（ELISA）法检测血清抗双链脱氧核糖核酸（anti-dsDNA）、抗单链 DNA、抗核抗体（ANA）和抗染色质水平。药物治疗 2 周后，通过基因芯片和基因集富集分析分别检测 miR-146a 模拟物和抑制剂对 MRL/lpr 小鼠肾组织的影响。24 周时处死小鼠，采集血样，采用 BUN 分析仪检测血尿素氮（BUN）水平。然后取肾组织，通过苏木精-伊红（HE）染色检测药物治疗对肾组织形态的影响。此外，采用实时定量聚合酶链反应（qRT-PCR）和 Western blot 分别检测药物治疗对肾组织中炎症因子和核因子-κB（NF-κB）信号通路 mRNA 水平的影响。
miR-146a 模拟物可显著减少尿蛋白的产生，且呈时间依赖性，同时可显著降低 24 周时的 BUN 水平。HE 染色结果显示，miR-146a 模拟物组肾小球损伤和肾血管损伤明显减轻。miR-146a 模拟物组血清抗 dsDNA、抗 ssDNA、抗染色质和 ANA 自身抗体水平显著降低，小鼠的生存时间显著延长。高通量基因表达芯片技术阐明，在 miR-146a 模拟物组中，NF-κB 正调控基因的表达呈下降趋势，而 NF-κB 负调控基因的表达呈上升趋势。miR-146a 模拟物可显著下调肾组织中 RELA、IRAK1、白细胞介素 1B（IL1B）和 IL-10 的表达水平。此外，Western blot 结果表明，miR-146a 模拟物抑制经典和非经典 NF-κB 信号通路。
miR-146a 通过调节经典和非经典 NF-κB 信号通路减少 MRL/lpr 小鼠 SLE 诱导的肾损伤。
