Cai Yan-Chun, Huang Qian, Wei Xiao-Li, Mei Ru-Huan, Sa Li-Na, Hu Xiao-Lan
Department of Physiology, School of Medicine, Zhejiang University, Hangzhou 310058, China.
The First Affiliated Hospital, Zhejiang University, Hangzhou 310006, China.
Sheng Li Xue Bao. 2019 Aug 25;71(4):575-580.
The aim of the present study was to investigate the effect of salidroside (Sal) on inflammatory activation induced by lipopolysaccharide (LPS) in the co-culture of rat alveolar macrophages (AM) NR 8383 and type II alveolar epithelial cells (AEC II) RLE-6TN. CCK-8 colorimetric method was used to detect cell proliferation percentage. The enzyme-linked immunosorbent assay (ELISA) was used to determine the content of tumor necrosis factor alpha (TNF-α), macrophage inflammatory protein-2 (MIP-2) and interleukin-10 (IL-10) in the supernatant. Western blot was used to examine the expression levels of phosphorylated AKT (p-AKT) and total AKT protein. The results showed that pretreatment of RLE-6TN cells or co-culture of RLE-6TN and NR 8383 cells with 32 and 128 µg/mL Sal for 1 h, followed by continuous culture for 24 h, significantly increased the cell proliferation (P < 0.05). Compared with control group, 32 and 128 µg/mL Sal pretreatment significantly increased the ratio of p-AKT/AKT in RLE-6TN cells (P < 0.05). Pretreatment of 32 µg/mL Sal not only inhibited the secretion of TNF-α and MIP-2 by NR 8383 cells induced by LPS (P < 0.05), but also enhanced the inhibitory effect of RLE-6TN and NR 8383 cells co-culture on the secretion of TNF-α and MIP-2 by NR 8383 cells induced by LPS (P < 0.05). In addition, 32 µg/mL Sal pretreatment promoted LPS-induced IL-10 secretion by NR 8383 cells (P < 0.05), and enhanced the promoting effect of co-culture of RLE-6TN and NR 8383 cells on the IL-10 secretion by LPS-induced NR 8383 cells (P < 0.05). In conclusion, Sal may directly inhibit LPS-induced inflammatory activation of AM (NR 8383), promote the proliferation of AEC II (RLE-6TN) through PI3K/AKT signaling pathway, and enhance the regulatory effect of AEC II on LPS-induced inflammatory activation of AM.
本研究旨在探讨红景天苷（Sal）对大鼠肺泡巨噬细胞（AM）NR 8383和II型肺泡上皮细胞（AEC II）RLE-6TN共培养体系中脂多糖（LPS）诱导的炎症激活的影响。采用CCK-8比色法检测细胞增殖率。酶联免疫吸附测定（ELISA）法测定上清液中肿瘤坏死因子α（TNF-α）、巨噬细胞炎性蛋白-2（MIP-2）和白细胞介素-10（IL-10）的含量。蛋白质免疫印迹法检测磷酸化AKT（p-AKT）和总AKT蛋白的表达水平。结果显示，用32和128 μg/mL的Sal预处理RLE-6TN细胞，或RLE-6TN与NR 8383细胞共培养1 h后，再持续培养24 h，细胞增殖显著增加（P<0.05）。与对照组相比，32和128 μg/mL的Sal预处理显著增加了RLE-6TN细胞中p-AKT/AKT的比值（P<0.05）。32 μg/mL的Sal预处理不仅抑制了LPS诱导的NR 8383细胞分泌TNF-α和MIP-2（P<0.05），还增强了RLE-6TN与NR 8383细胞共培养对LPS诱导的NR 8383细胞分泌TNF-α和MIP-2的抑制作用（P<0.05）。此外，32 μg/mL的Sal预处理促进了LPS诱导的NR 8383细胞分泌IL-10（P<0.05），并增强了RLE-6TN与NR 8383细胞共培养对LPS诱导的NR 8383细胞分泌IL-10的促进作用（P<0.05）。综上所述，Sal可能直接抑制LPS诱导的AM（NR 8383）炎症激活，通过PI3K/AKT信号通路促进AEC II（RLE-6TN）增殖，并增强AEC II对LPS诱导的AM炎症激活的调节作用。
