Abstract：
OBJECTIVE: This study aimed to determine the influence of miR-1297 on kidney injury in rats with diabetic nephropathy (DN) and its causal role.
METHODS: 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. miR-1297 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.
RESULTS: AntagomiR-1297 reduced BUN (p = 0.005), SCr (p = 0.012), and urine protein (p < 0.001) levels and improved kidney tissue morphology. It prevented renal interstitial fibrosis by decreasing FN, Col I, and α-SMA protein levels (all p < 0.001). AntagomiR-1297 increased SOD (p = 0.001) and GSH-Px (p = 0.002) levels. Additionally, it reduced levels of cell inflammatory factors, including TNF-α, IL-6, and IL-1β (all p < 0.001), and alleviated apoptosis (p < 0.001) in rat kidney tissue with DN. miR-1297 was pinpointed as a target for PTEN. AntagomiR-1297 increased PTEN expression and suppressed PI3K and AKT phosphorylation (all p < 0.001).
CONCLUSIONS: AntagomiR-1297 can mitigate renal fibrosis, renal inflammation, apoptosis, and oxidative stress levels through the PTEN/PI3K/AKT pathway.
METHODS: 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. miR-1297 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.
RESULTS: AntagomiR-1297 reduced BUN (p = 0.005), SCr (p = 0.012), and urine protein (p < 0.001) levels and improved kidney tissue morphology. It prevented renal interstitial fibrosis by decreasing FN, Col I, and α-SMA protein levels (all p < 0.001). AntagomiR-1297 increased SOD (p = 0.001) and GSH-Px (p = 0.002) levels. Additionally, it reduced levels of cell inflammatory factors, including TNF-α, IL-6, and IL-1β (all p < 0.001), and alleviated apoptosis (p < 0.001) in rat kidney tissue with DN. miR-1297 was pinpointed as a target for PTEN. AntagomiR-1297 increased PTEN expression and suppressed PI3K and AKT phosphorylation (all p < 0.001).
CONCLUSIONS: AntagomiR-1297 can mitigate renal fibrosis, renal inflammation, apoptosis, and oxidative stress levels through the PTEN/PI3K/AKT pathway.
摘要：
目的：本研究旨在确定miR-1297对糖尿病肾病（DN）大鼠肾损伤的影响及其因果作用。
方法：通过右肾切除和腹腔注射链脲佐菌素（STZ）建立DN大鼠模型。假大鼠没有进行右肾切除或STZ注射。将DN大鼠分为DN模型组和antagomiR-1297治疗组。使用苏木精和伊红染色观察肾脏形态。肾功能指标,包括血尿素氮（BUN），血清肌酐（SCr），和尿蛋白，使用试剂盒测量。肿瘤坏死因子-α(TNF-α)水平,白细胞介素（IL）-6，IL-1β，超氧化物歧化酶（SOD），通过酶联免疫吸附试验（ELISA）测定谷胱甘肽过氧化物酶（GSH-Px）。纤维蛋白(FN),I型胶原蛋白（ColI），通过蛋白质印迹和实时逆转录聚合酶链反应评估α-平滑肌肌动蛋白（α-SMA）。使用末端脱氧核苷酸转移酶dUTP缺口末端标记染色检测细胞凋亡。使用生物信息学软件预测miR-1297靶标，并通过荧光素酶报告基因测定进行验证。通过蛋白质印迹分析了10号染色体上缺失的磷酸酶和张力蛋白同源物（PTEN）/磷酸肌醇3-激酶（PI3K）/蛋白激酶B（AKT）途径的表达。
结果：AntagomiR-1297降低了BUN（p=0.005），SCr（p=0.012），尿蛋白(p<0.001)水平和改善肾脏组织形态。它通过降低FN来预防肾间质纤维化，上校I,和α-SMA蛋白水平（所有p<0.001）。AntagomiR-1297增加SOD（p=0.001）和GSH-Px（p=0.002）水平。此外,它降低了细胞炎症因子的水平，包括TNF-α,IL-6和IL-1β（所有p<0.001），减轻了DN大鼠肾组织的凋亡（p<0.001）。miR-1297被确定为PTEN的靶标。AntagomiR-1297增加PTEN表达并抑制PI3K和AKT磷酸化(所有p<0.001)。
结论：AntagomiR-1297可以减轻肾脏纤维化，肾脏炎症，凋亡，和氧化应激水平通过PTEN/PI3K/AKT途径。
方法：通过右肾切除和腹腔注射链脲佐菌素（STZ）建立DN大鼠模型。假大鼠没有进行右肾切除或STZ注射。将DN大鼠分为DN模型组和antagomiR-1297治疗组。使用苏木精和伊红染色观察肾脏形态。肾功能指标,包括血尿素氮（BUN），血清肌酐（SCr），和尿蛋白，使用试剂盒测量。肿瘤坏死因子-α(TNF-α)水平,白细胞介素（IL）-6，IL-1β，超氧化物歧化酶（SOD），通过酶联免疫吸附试验（ELISA）测定谷胱甘肽过氧化物酶（GSH-Px）。纤维蛋白(FN),I型胶原蛋白（ColI），通过蛋白质印迹和实时逆转录聚合酶链反应评估α-平滑肌肌动蛋白（α-SMA）。使用末端脱氧核苷酸转移酶dUTP缺口末端标记染色检测细胞凋亡。使用生物信息学软件预测miR-1297靶标，并通过荧光素酶报告基因测定进行验证。通过蛋白质印迹分析了10号染色体上缺失的磷酸酶和张力蛋白同源物（PTEN）/磷酸肌醇3-激酶（PI3K）/蛋白激酶B（AKT）途径的表达。
结果：AntagomiR-1297降低了BUN（p=0.005），SCr（p=0.012），尿蛋白(p<0.001)水平和改善肾脏组织形态。它通过降低FN来预防肾间质纤维化，上校I,和α-SMA蛋白水平（所有p<0.001）。AntagomiR-1297增加SOD（p=0.001）和GSH-Px（p=0.002）水平。此外,它降低了细胞炎症因子的水平，包括TNF-α,IL-6和IL-1β（所有p<0.001），减轻了DN大鼠肾组织的凋亡（p<0.001）。miR-1297被确定为PTEN的靶标。AntagomiR-1297增加PTEN表达并抑制PI3K和AKT磷酸化(所有p<0.001)。
结论：AntagomiR-1297可以减轻肾脏纤维化，肾脏炎症，凋亡，和氧化应激水平通过PTEN/PI3K/AKT途径。
