Cai Songtao, Sun Jingtao, Wei Xuan
Department of Arthropathy, Zhengzhou Orthopaedic Hospital, Zhengzhou 450052, China.
Department of Arthropathy, Zhengzhou Orthopaedic Hospital, Zhengzhou 450052, China. *Corresponding author, E-mail:
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2018 Nov;34(11):989-993.
Objective To investigate the effect of Lycium barbarum polysaccharide (LBP) on the proliferation and immune-related cytokines in osteoarthritis (OA) chondrocytes. Methods We isolated chondrocytes from OA samples, and then detected cell proliferation of OA chondrocytes treated with LBP at 0, 100, 200, 400 and 800 μg/mL by MTT assay. OA chondrocytes were treated with 400 μg/mL LBP which was determined to be the optimal concentration by MTT assay. Real-time fluorescence quantitative PCR and Western blot analysis were performed to detect the expression of induced nitric oxide synthase (iNOS), growth transformation factor beta (TGF-β) and nuclear factor-κBp65 (NF-κBp65). And the levels of interleukin 1 beta (IL-1β) and tumor necrosis factor alpha (TNF-α) in the supernatant were detected by ELISA. Results Cell viability of OA chondrocytes treated with LBP at 200, 400 or 800 μg/mL was significantly lower than that at 0 μg/mL, and cell viability of 400 μg/mL group and 800 μg/mL group was lower than that of 200 μg/mL group. There was no significant difference in the cell viability between the 400 μg/mL group and 800 μg/mL group. So 400 μg/mL LBP was used for subsequent experiments. Compared with the control group, the levels of IL-1β and TNF-α in the supernatant of OA chondrocytes after treated with 400 μg/mL LBP decreased; the expression of iNOS mRNA and protein were down-regulated; the expression of TGF-β was up-regulated; and the expression level of NF-κBp65 protein was reduced. Conclusion LBP could reduce the levels of immune-related cytokines in OA chondrocytes and inhibit the NF-κB signaling pathway, which is instrumental in improving the OA injury.
目的 探讨枸杞多糖（LBP）对骨关节炎（OA）软骨细胞增殖及免疫相关细胞因子的影响。方法 从OA样本中分离软骨细胞，采用MTT法检测0、100、200、400和800 μg/mL LBP处理的OA软骨细胞的增殖情况。用MTT法确定最佳浓度为400 μg/mL的LBP处理OA软骨细胞。采用实时荧光定量PCR和蛋白质印迹分析检测诱导型一氧化氮合酶（iNOS）、生长转化因子β（TGF-β）和核因子κB p65（NF-κB p65）的表达。采用酶联免疫吸附测定法检测上清液中白细胞介素1β（IL-1β）和肿瘤坏死因子α（TNF-α）的水平。结果 200、400或800 μg/mL LBP处理的OA软骨细胞活力明显低于0 μg/mL组，400 μg/mL组和800 μg/mL组细胞活力低于200 μg/mL组。400 μg/mL组和800 μg/mL组细胞活力差异无统计学意义。因此，后续实验采用400 μg/mL LBP。与对照组相比，400 μg/mL LBP处理后的OA软骨细胞上清液中IL-1β和TNF-α水平降低；iNOS mRNA和蛋白表达下调；TGF-β表达上调；NF-κB p65蛋白表达水平降低。结论 LBP可降低OA软骨细胞中免疫相关细胞因子水平，抑制NF-κB信号通路，有助于改善OA损伤。
