Emergency Department, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou City, 310006, Zhejiang Province, China.
Department of Cardiology, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou City, 310006, Zhejiang Province, China.
Sci Rep. 2020 Nov 27;10(1):20750. doi: 10.1038/s41598-020-77448-6.
The purpose of this study was to explore the investigative mechanism of salidroside (SAL) on LPS-induced acute lung injury (ALI)/acute respiratory distress syndrome (ARDS). The exosomes from RLE-6TN are extracted and identified by transmission electron microscopy, particle size analysis and protein marker detection, and co-cultured with NR8383 cells. The ALI/ARDS model of SD rats was established by LPS (10 mg/kg) intratracheal instillation. Following a four-hour intratracheal instillation of LPS, 50 μl of RLE-6TN exosomes were injected through the tail vein. After that, SAL and miR-146a antagomir were injected into the tail vein for 72 h, respectively. As the changes of HE stain, body weight and ALI score are observed. The expression of miR-146a, TLR4, NF-kB, IRAK1, TRAF6 and their related proteins were detected by RT-PCR and Western blot, respectively. TNF-α, IL-6, IL-8 and IL-1 β inflammatory factors were detected by ELISA. The expression of miR-146a, NF-kB, IRAK, TRAF6 and related inflammatory factors in LPS-induced NR8383 was significantly higher than that in the control group, while SAL has greatly reduced the expression of TLR4 mediated NF-kB inflammatory pathway and related inflammatory factors. SAL can significantly improve the LPS-induced lung morphological abnormalities, slowed down the rate of weight loss in rats, and reducing the ALI score. The expression trend of NF-kB, IRAK, TRAF6 and related inflammatory factors in rats' lung tissues was consistent with that in NR8383 cells. SAL has a protective effect on ALI/ARDS caused by sepsis, which is likely to be developed to a potential treatment for the disease. To sum up, this study provides a new theoretical basis for the treatment of ALI/ARDS with SAL.
本研究旨在探讨红景天苷（SAL）对脂多糖（LPS）诱导的急性肺损伤（ALI）/急性呼吸窘迫综合征（ARDS）的作用机制。通过透射电子显微镜、粒径分析和蛋白标志物检测提取并鉴定 RLE-6TN 细胞来源的外泌体，并与 NR8383 细胞共培养。通过 LPS（10mg/kg）气管内滴注建立 SD 大鼠 ALI/ARDS 模型。LPS 气管内滴注 4 小时后，经尾静脉注入 50μl RLE-6TN 外泌体。随后，尾静脉分别注入 SAL 和 miR-146a 拮抗剂，共 72 小时。观察 HE 染色、体重和 ALI 评分的变化。采用 RT-PCR 和 Western blot 分别检测 miR-146a、TLR4、NF-κB、IRAK1、TRAF6 及其相关蛋白的表达。ELISA 法检测 TNF-α、IL-6、IL-8 和 IL-1β炎症因子。与对照组相比，LPS 诱导的 NR8383 中 miR-146a、NF-κB、IRAK、TRAF6 及其相关炎症因子的表达明显升高，而 SAL 则大大降低了 TLR4 介导的 NF-κB 炎症通路及其相关炎症因子的表达。SAL 能明显改善 LPS 诱导的肺形态学异常，减缓大鼠体重减轻率，降低 ALI 评分。大鼠肺组织中 NF-κB、IRAK、TRAF6 及相关炎症因子的表达趋势与 NR8383 细胞一致。SAL 对脓毒症引起的 ALI/ARDS 具有保护作用，可能为该疾病的治疗提供新的理论依据。综上所述，本研究为 SAL 治疗 ALI/ARDS 提供了新的理论依据。
