Department of Cardiothoracic Surgery, Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing, Jiangsu Province, China.
Department of Cardiothoracic Surgery, Jiangsu Province Academy of Traditional Chinese Medicine, Nanjing, Jiangsu Province, China.
Bioengineered. 2021 Dec;12(1):7747-7754. doi: 10.1080/21655979.2021.1982845.
We aimed to explore the role of miR-21-5p in the inhibitory effects of astragaloside IV (As-IV) on hypoxia/reoxygenation injury-induced apoptosis of type II alveolar epithelial cells. Rat type II alveolar epithelial cells RLE-6TN were cultured and randomly divided into control (C), hypoxia/reoxygenation injury (H/R), As-IV and miR-21-5p-siRNA + As-IV groups (n = 6). H/R model was established by 24 h of hypoxia and 4 h of reoxygenation. As-IV group was given 1 nmol/L As-IV and incubated for 1 h before modeling. MiR-21-5p-siRNA + As-IV group was transfected with 50 nmol/L miR-21-5p-siRNA. After 48 h, they were incubated with 1 nmol/L As-IV for 1 h before modeling. Cell viability was detected by cell counting kit-8 assay, and apoptosis rate was detected by flow cytometry. The expression levels of TLR4 and NF-κB were measured by immunofluorescence assay. The targeting relationship between miR-21-5p and TLR4 was determined by luciferase assay. Compared with H/R group, the cell viability, miR-21-5p, bax and cleaved caspase-3 expressions of As-IV group increased, apoptosis rate and Bcl-2 expression decreased, and TLR4 and NF-κB expressions were down-regulated (P < 0.05). Compared with As-IV group, the cell viability, miR-21-5p, bax and cleaved caspase-3 expressions of miR-21-5p-siRNA + As-IV group decreased, apoptosis rate and Bcl-2 expression increased, and the expressions of TLR4 and NF-κB were up-regulated (P < 0.05). As-IV up-regulates miR-21-5p expression, inhibits the TLR4/NF-κB signaling pathway and suppresses the apoptosis of type II alveolar epithelial cells during hypoxia/reoxygenation injury.
我们旨在探讨 miR-21-5p 在黄芪甲苷 (As-IV) 抑制缺氧/复氧损伤诱导的 II 型肺泡上皮细胞凋亡中的作用。培养大鼠 II 型肺泡上皮细胞 RLE-6TN，随机分为对照组 (C)、缺氧/复氧损伤 (H/R)、As-IV 和 miR-21-5p-siRNA + As-IV 组 (n = 6)。通过 24 h 缺氧和 4 h 复氧建立 H/R 模型。As-IV 组在建模前给予 1 nmol/L As-IV 孵育 1 h。miR-21-5p-siRNA + As-IV 组转染 50 nmol/L miR-21-5p-siRNA。建模前，孵育 1 nmol/L As-IV 1 h。采用细胞计数试剂盒-8 法检测细胞活力，流式细胞术检测细胞凋亡率。免疫荧光法检测 TLR4 和 NF-κB 的表达水平。通过荧光素酶报告基因检测 miR-21-5p 与 TLR4 的靶向关系。与 H/R 组相比，As-IV 组细胞活力、miR-21-5p、bax 和 cleaved caspase-3 表达增加，细胞凋亡率和 Bcl-2 表达降低，TLR4 和 NF-κB 表达下调 (P < 0.05)。与 As-IV 组相比，miR-21-5p-siRNA + As-IV 组细胞活力、miR-21-5p、bax 和 cleaved caspase-3 表达降低，细胞凋亡率和 Bcl-2 表达增加，TLR4 和 NF-κB 表达上调 (P < 0.05)。As-IV 上调 miR-21-5p 表达，抑制 TLR4/NF-κB 信号通路，抑制缺氧/复氧损伤时 II 型肺泡上皮细胞凋亡。
