Department of Orthopaedic Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700-8558, Japan.
Department of Orthopaedic Surgery, Okayama University Hospital, Okayama 700-8558, Japan.
Int J Mol Sci. 2021 Sep 10;22(18):9781. doi: 10.3390/ijms22189781.
We investigated the effects of adipose-derived extract (AE) on cultured chondrocytes and in vivo cartilage destruction. AE was prepared from human adipose tissues using a nonenzymatic approach. Cultured human chondrocytes were stimulated with interleukin-1 beta (IL-1β) with or without different concentrations of AE. The effects of co-treatment with AE on intracellular signaling pathways and their downstream gene and protein expressions were examined using real-time PCR, Western blotting, and immunofluorescence staining. Rat AE prepared from inguinal adipose tissues was intra-articularly delivered to the knee joints of rats with experimental osteoarthritis (OA), and the effect of AE on cartilage destruction was evaluated histologically. In vitro, co-treatment with IL-1β combined with AE reduced activation of the p38 and ERK mitogen-activated protein kinase (MAPK) pathway and nuclear translocation of the p65 subunit of nuclear factor-kappa B (NF-κB), and subsequently downregulated the expressions of matrix metalloproteinase (MMP)-1, MMP-3, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)-4, IL-6, and IL-8, whereas it markedly upregulated the expression of IL-1 receptor type 2 (IL-1R2) in chondrocytes. Intra-articular injection of homologous AE significantly ameliorated cartilage destruction six weeks postoperatively in the rat OA model. These results suggested that AE may exert a chondroprotective effect, at least in part, through modulation of the IL-1β-induced inflammatory signaling pathway by upregulation of IL-1R2 expression.
我们研究了脂肪衍生提取物 (AE) 对培养的软骨细胞和体内软骨破坏的影响。AE 是使用非酶方法从人脂肪组织中制备的。用白细胞介素-1β (IL-1β) 刺激培养的人软骨细胞,并用或不用不同浓度的 AE 进行共处理。使用实时 PCR、Western blot 和免疫荧光染色检查 AE 对细胞内信号通路及其下游基因和蛋白表达的共处理效应。从腹股沟脂肪组织制备的大鼠 AE 被关节内递送到实验性骨关节炎 (OA) 的膝关节,并评估 AE 对软骨破坏的影响。在体外,IL-1β与 AE 联合共处理可减少 p38 和 ERK 丝裂原活化蛋白激酶 (MAPK) 通路的激活和核因子-κB (NF-κB) p65 亚单位的核易位,随后下调基质金属蛋白酶 (MMP)-1、MMP-3、MMP-13、金属蛋白酶与血小板反应蛋白基序 (ADAMTS)-4、IL-6 和 IL-8 的表达,而明显上调软骨细胞中白细胞介素-1 受体类型 2 (IL-1R2) 的表达。关节内注射同源 AE 可显著改善大鼠 OA 模型术后六周的软骨破坏。这些结果表明,AE 至少部分通过上调 IL-1R2 表达来调节 IL-1β 诱导的炎症信号通路,从而发挥软骨保护作用。