红景天苷对脂多糖诱导大鼠肺泡巨噬细胞和II型肺泡上皮细胞共培养炎性介质分泌的影响
蔡艳春¹, 黄倩², 危晓莉¹, 梅汝焕¹, 撒丽娜¹, 胡晓兰^1,*
1浙江大学医学院生理学系，杭州 310058；²浙江大学第一附属医院，杭州 310006
摘要
本研究旨在探讨红景天苷(salidroside, Sal)对脂多糖(lipopolysaccharide, LPS)诱导大鼠肺泡巨噬细胞NR 8383和II型肺泡上皮细胞RLE-6TN共培养炎性活化的影响。CCK-8比色法检测细胞增殖百分率，Western blot检测磷酸化AKT (p-AKT)和总AKT蛋白表达，酶联免疫吸附法测定细胞培养上清中肿瘤坏死因子α (tumor necrosis factor α, TNF-α)、巨噬细胞炎性蛋白2 (macrophage inflammatory protein-2, MIP-2)和白介素10 (interleukin-10, IL-10)的含量。结果显示：与对照组相比，32和128 μg/mL Sal预处理RLE-6TN细胞或共培养RLE-6TN和NR 8383细胞1 h后继续培养24 h，细胞增殖百分率显著增加(P < 0.05)；与对照组相比，32和128 μg/mL Sal预处理RLE-6TN细胞，p-AKT/AKT蛋白比值显著增加(P < 0.05)。32 μg/mL Sal预处理不仅抑制LPS诱导NR 8383细胞分泌TNF-α和MIP-2 (P < 0.05)，而且加强RLE-6TN和NR 8383细胞共培养对LPS诱导NR 8383细胞分泌TNF-α和MIP-2的抑制作用(P < 0.05)。此外，32 μg/mL Sal预处理能促进LPS诱导NR 8383细胞分泌IL-10 (P < 0.05)，并能加强RLE-6TN和NR 8383细胞共培养对LPS诱导NR 8383细胞分泌IL-10的促进作用(P < 0.05)。以上结果提示，Sal不仅能直接抑制LPS诱导的NR 8383炎性活化，还可能通过PI3K/AKT信号通路促进RLE-6TN增殖，参与II型肺泡上皮细胞对LPS诱导肺泡巨噬细胞炎性活化的调节作用。
关键词： 红景天苷; 脂多糖; 肺泡巨噬细胞; II型肺泡上皮细胞
分类号：Q291

Effects of salidroside on the secretion of inflammatory mediators induced by lipopolysaccharide in the co-culture of rat alveolar macrophages and type II alveolar epithelial cells
CAI Yan-Chun¹, HUANG Qian², WEI Xiao-Li¹, MEI Ru-Huan¹, SA Li-Na¹, HU Xiao-Lan^1,*
1Department of Physiology, School of Medicine, Zhejiang University, Hangzhou 310058, China；²he First Affiliated Hospital, Zhejiang University, Hangzhou 310006, China
Abstract
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.


Key words: salidroside; lipopolysaccharide; alveolar macrophages; type II alveolar epithelial cells

收稿日期：2018-11-18　　录用日期：2019-03-12
通讯作者：胡晓兰　　E-mail: huxiaolan@zju.edu.cn
DOI: 10.13294/j.aps.2019.0024
引用本文：
蔡艳春, 黄倩, 危晓莉, 梅汝焕, 撒丽娜, 胡晓兰. 红景天苷对脂多糖诱导大鼠肺泡巨噬细胞和II型肺泡上皮细胞共培养炎性介质分泌的影响[J]. 生理学报 2019; 71 (4): 575-580.
CAI Yan-Chun, HUANG Qian, WEI Xiao-Li, MEI Ru-Huan, SA Li-Na, HU Xiao-Lan. Effects of salidroside on the secretion of inflammatory mediators induced by lipopolysaccharide in the co-culture of rat alveolar macrophages and type II alveolar epithelial cells. Acta Physiol Sin 2019; 71 (4): 575-580 (in Chinese with English abstract).



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