Ci Binbin, Wang Wenjun, Ni Yongan
Department of Orthopedics, The Wendeng Osteopath Hospital, Weihai, China.
Department of Emergency, The Wendeng Osteopath Hospital, Weihai, China.
Pak J Pharm Sci. 2018 Nov;31(6(Special)):2791-2798.
To study the mechanism and inhibitory effect of Saposhnikovia divaricata polysaccharide (SDP) on fibroblast-like synoviocytes (FLS) isolated from rheumatoid arthritis rat model. Rheumatoid arthritis rat model was established by the classical composite factors including wind, cold, damp plus biological agents. The synovial tissues were digested with trypsin to isolate FLS cells. The different dosage of SDP was applied in culture. The cell viability was evaluated by MTT assay and the apoptosis was determined by analytic flow cytometry. The expression change of p53 gene was monitored by RT-PCR method. The production of secretory inflammation factors TNF-α and IL-1β were determined by ELISA. The proliferative and apoptotic proteins such as Bcl-2, Bax, Caspase-3, MMP-1, MMP-3, P53 were measured by western blotting. Our data demonstrated that treatment with high concentration of SDP could enhance the expression of P53 at both mRNA (P<0.05) and protein (P<0.05) level, inhibit the secretion of TNF-α (P<0.05) and IL-1β (P<0.05). Simultaneously, the Bcl-2/Bax ratio and level of MMP-1, MMP-3 was significantly decreased, and apoptotic marker caspase-3 protein was increased. In addition, the FACS analysis consistently consolidated the apoptosis-inducing effect of SDP on RAFLS. SDP could significantly inhibit dysplasia of RAFLS via modulation of p53 expression and suppression of inflammatory factors, which suggested a potential therapeutic value for rheumatoid arthritis.
研究防风多糖(SDP)对类风湿关节炎大鼠模型分离的成纤维样滑膜细胞(FLS)的作用机制及抑制作用。采用风、寒、湿加生物制剂等经典复合因素建立类风湿关节炎大鼠模型。用胰蛋白酶消化滑膜组织以分离FLS细胞。在培养中应用不同剂量的SDP。通过MTT法评估细胞活力,通过流式细胞术分析确定细胞凋亡。用RT-PCR法监测p53基因的表达变化。用ELISA法测定分泌性炎症因子TNF-α和IL-1β的产生。通过蛋白质印迹法检测Bcl-2、Bax、Caspase-3、MMP-1、MMP-3、P53等增殖和凋亡蛋白。我们的数据表明,高浓度SDP处理可在mRNA(P<0.05)和蛋白质(P<0.05)水平增强P53的表达,抑制TNF-α(P<0.05)和IL-1β(P<0.05)的分泌。同时,Bcl-2/Bax比值以及MMP-1、MMP-3水平显著降低,凋亡标志物caspase-3蛋白增加。此外,流式细胞术分析一致证实了SDP对类风湿关节炎成纤维样滑膜细胞(RAFLS)的诱导凋亡作用。SDP可通过调节p53表达和抑制炎症因子显著抑制RAFLS的发育异常,这提示其对类风湿关节炎具有潜在的治疗价值。