Blood Purification Center, Qingdao Central Hospital, 266042 Qingdao, Shandong, China.
Department of Nephrology, Qingdao Central Hospital, 266042 Qingdao, Shandong, China.
Arch Esp Urol. 2024 Mar;77(2):183-192. doi: 10.56434/j.arch.esp.urol.20247702.24.
This study aimed to determine the influence of on kidney injury in rats with diabetic nephropathy (DN) and its causal role.
A DN rat model was established through right kidney resection and intraperitoneal injection of streptozotocin (STZ). Sham rats did not undergo right kidney resection or STZ injection. The DN rats were divided into the DN model and antagomiR-1297 treatment groups. Kidney morphology was observed using hematoxylin and eosin staining. Renal function indices, including blood urea nitrogen (BUN), serum creatinine (SCr), and urinary protein, were measured using kits. Levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-6, IL-1β, superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) were determined through enzyme-linked immunosorbent assay (ELISA). Fibrin (FN), collagen type I (Col I), and α-smooth muscle actin (α-SMA) were assessed through western blotting and real-time reverse transcription-polymerase chain reaction. Apoptosis was detected using terminal deoxynucleotidyl transferase dUTP nick end labeling staining. targets were predicted using bioinformatic software and verified through luciferase reporter assay. Phosphatase and tensin homolog deleted on chromosome 10 (PTEN)/phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) pathway expression was analyzed through western blotting.
AntagomiR-1297 reduced BUN ( = 0.005), SCr ( = 0.012), and urine protein ( < 0.001) levels and improved kidney tissue morphology. It prevented renal interstitial fibrosis by decreasing FN, Col I, and α-SMA protein levels (all < 0.001). AntagomiR-1297 increased SOD ( = 0.001) and GSH-Px ( = 0.002) levels. Additionally, it reduced levels of cell inflammatory factors, including TNF-α, IL-6, and IL-1β (all < 0.001), and alleviated apoptosis ( < 0.001) in rat kidney tissue with DN. was pinpointed as a target for PTEN. AntagomiR-1297 increased PTEN expression and suppressed PI3K and AKT phosphorylation (all < 0.001).
AntagomiR-1297 can mitigate renal fibrosis, renal inflammation, apoptosis, and oxidative stress levels through the PTEN/PI3K/AKT pathway.
本研究旨在探讨 miR-1297 对糖尿病肾病（DN）大鼠肾损伤的影响及其因果关系。
通过右肾切除术和链脲佐菌素（STZ）腹腔注射建立 DN 大鼠模型。假手术大鼠未进行右肾切除术或 STZ 注射。将 DN 大鼠分为 DN 模型组和 antagomiR-1297 治疗组。采用苏木精-伊红染色观察肾脏形态。采用试剂盒测定血尿素氮（BUN）、血清肌酐（SCr）和尿蛋白等肾功能指标。采用酶联免疫吸附试验（ELISA）测定肿瘤坏死因子-α（TNF-α）、白细胞介素（IL）-6、IL-1β、超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）水平。采用 Western blot 和实时逆转录-聚合酶链反应检测纤维连接蛋白（FN）、胶原 I（Col I）和α-平滑肌肌动蛋白（α-SMA）。采用末端脱氧核苷酸转移酶 dUTP 缺口末端标记染色法检测细胞凋亡。采用生物信息学软件预测 miR-1297 的靶基因，并通过荧光素酶报告基因实验进行验证。采用 Western blot 分析磷酸酶和张力蛋白同源物缺失的第 10 号染色体（PTEN）/磷酸肌醇 3-激酶（PI3K）/蛋白激酶 B（AKT）通路表达。
antagomiR-1297 降低了 BUN（ = 0.005）、SCr（ = 0.012）和尿蛋白（ < 0.001）水平，改善了肾脏组织形态。它通过降低 FN、Col I 和 α-SMA 蛋白水平（均 < 0.001）来防止肾间质纤维化。antagomiR-1297 增加了 SOD（ = 0.001）和 GSH-Px（ = 0.002）水平。此外，它降低了 TNF-α、IL-6 和 IL-1β 等细胞炎症因子的水平（均 < 0.001），并减轻了糖尿病肾病大鼠肾脏组织的细胞凋亡（ < 0.001）。PTEN 被确定为 miR-1297 的靶基因。antagomiR-1297 增加了 PTEN 的表达，并抑制了 PI3K 和 AKT 的磷酸化（均 < 0.001）。
antagomiR-1297 可通过 PTEN/PI3K/AKT 通路减轻肾纤维化、肾炎症、细胞凋亡和氧化应激水平。
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