Hsueh M-F, Bolognesi M P, Wellman S S, Kraus V B
Duke Molecular Physiology Institute, Durham, NC, USA; Department. of Orthopaedic Surgery, Duke University Medical Center, Durham, NC, USA.
Department. of Orthopaedic Surgery, Duke University Medical Center, Durham, NC, USA.
Osteoarthritis Cartilage. 2020 May;28(5):639-645. doi: 10.1016/j.joca.2020.01.013. Epub 2020 Feb 4.
To evaluate the anti-inflammatory effects of clinically relevant naproxen sodium (Nx) concentrations on human monocyte-derived macrophages in a controlled in vitro system and human primary synovial fluid (SF) cells.
Using phorbol 12-myristate 13-acetate, THP-1 human monocytic cells were differentiated into mature monocyte-derived macrophages in vitro then treated with Nx pre- or post-activating an inflammatory response with lipopolysaccharide (LPS) and hyaluronan (HA) fragments (n = 8/group). Cell culture supernatants were assessed for NF-κB activity and prostaglandin E (PGE), indicating cyclooxygenase enzyme activity. Under Duke IRB approval, primary human SF cells were collected at the time of knee joint replacement (n = 19 individuals) for osteoarthritis (OA), and cultured with LPS, HA and Nx; SF cells were characterized by polychromatic flow cytometry for cell surface markers and intracellular cytokines.
Compared to placebo treatment of THP-1 cells, low dose Nx (corresponding 27.5-440 mg/L orally) added both pre- and post-activation with LPS/HA, significantly reduced NF-κB activity and PGE2: mean reduction to 73%, 61%, 17% and 10% of placebo, respectively. LPS/HA treatment of primary OA SF cells significantly increased the number of IL-1β producing primary monocytes and macrophages, and by 24 h the overall production of secreted cytokines (IL-1β, IL-6, IL8, and TNF-α). Low dose Nx reduced the percentage of IL-1β producing primary monocytes and macrophages.
LPS/HA induced inflammation of THP-1 monocytic and primary human SF cells. Low dose Nx both prevented and reduced inflammatory responses of a human monocytic cell line and reduced IL-1β production by primary human SF monocytes and macrophages.
在可控的体外系统和人原发性滑液（SF）细胞中，评估临床相关浓度的萘普生钠（Nx）对人单核细胞衍生巨噬细胞的抗炎作用。
使用佛波醇12 - 肉豆蔻酸酯13 - 乙酸酯，将THP - 1人单核细胞在体外分化为成熟的单核细胞衍生巨噬细胞，然后在脂多糖（LPS）和透明质酸（HA）片段激活炎症反应之前或之后用Nx处理（每组n = 8）。评估细胞培养上清液中的NF - κB活性和前列腺素E（PGE），以指示环氧化酶活性。在杜克大学机构审查委员会批准下，在膝关节置换时收集原发性人SF细胞（n = 19例）用于骨关节炎（OA），并与LPS、HA和Nx一起培养；通过多色流式细胞术对SF细胞进行细胞表面标志物和细胞内细胞因子的表征。
与THP - 1细胞的安慰剂处理相比，在LPS/HA激活之前和之后添加低剂量Nx（相当于口服27.5 - 440 mg/L），显著降低了NF - κB活性和PGE2：分别平均降低至安慰剂的73%、61%、17%和10%。LPS/HA处理原发性OA SF细胞显著增加了产生IL - 1β的原发性单核细胞和巨噬细胞的数量，并且在24小时时分泌细胞因子（IL - 1β、IL - 6、IL8和TNF - α）的总体产量增加。低剂量Nx降低了产生IL - 1β的原发性单核细胞和巨噬细胞的百分比。
LPS/HA诱导THP - 1单核细胞和原发性人SF细胞发生炎症。低剂量Nx既预防又减少了人单核细胞系的炎症反应，并降低了原发性人SF单核细胞和巨噬细胞产生IL - 1β的量。
