Sun Yue, Ding Cong-zhu, Yao Yao
Department of Rheumatology and Immunology, Nanjing Drum Tower Hospital, Nanjing University of Traditional Chinese Medicine, Nanjing.
Zhongguo Zhong Xi Yi Jie He Za Zhi. 2012 Aug;32(8):1107-11.
To investigate the effects of sinomenine (SIN) and methotrexate (MTX) on the proliferation and apoptosis of in vitro cultured fibroblast-like synoviocytes (FLS) in rheumatoid arthritis (RA) patients, as well as the expression of osteoclast differentiation factor in FLS.
FLS were isolated from the synovium of RA patients and cultured in vitro. FLS were incubated with different concentrations of SIN and MTX respectively or combined: 0.001, 0.010, 0.100, 1.000 mg/mL SIN; 0.001, 0.010, 0.100, 1.000 mg/mL MTX; 0.001 mg/mL SIN + 0.001 mg/mL MTX, 0.010 mg/mL SIN + 0.010 mg/mL MTX, 0.100 mg/mL SIN + 0.100 mg/mL MTX, 1.000 mg/mL SIN + 1.000 mg/mL MTX, namely SIN1, 2, 3, 4 groups; MTX1, 2, 3, 4 groups and the combination 1, 2, 3, 4 groups. The medium without drugs was used as a control group. There was a total of 13 groups, each group with 3 complex holes. MTT was applied to detect the growth of FLS. The flow cytometry was applied to detect the apoptosis of FLS. The expressions of FLS receptor activator of nuclear factor kappa B ligand (RANKL) mRNA and osteoprotegerin (OPG) mRNA were observed by semi-quantitative RT-PCR.
Compared with the control group, RA FLS proliferation OD values of all the drug groups were lower (P < 0.05). The RA FLS apoptosis OD value of the combination 3 group increased, the OPG mRNA expression increased, the expression of RANKL mRNA decreased with statistical difference (P < 0.05). The RA proliferation OD values of the SIN3 group and the MTX3 group increased when compared with the combination 3 group (P < 0.05).
SIN and MTX had synergistic effects in inhibiting FLS. This might be one of the mechanisms for inhibiting RA bone damage.
探讨青藤碱（SIN）与甲氨蝶呤（MTX）对类风湿关节炎（RA）患者体外培养的成纤维样滑膜细胞（FLS）增殖及凋亡的影响，以及对FLS中破骨细胞分化因子表达的影响。
从RA患者滑膜中分离FLS并进行体外培养。分别用不同浓度的SIN和MTX单独或联合孵育FLS：0.001、0.010、0.100、1.000 mg/mL SIN；0.001、0.010、0.100、1.000 mg/mL MTX；0.001 mg/mL SIN + 0.001 mg/mL MTX、0.010 mg/mL SIN + 0.010 mg/mL MTX、0.100 mg/mL SIN + 0.100 mg/mL MTX、1.000 mg/mL SIN + 1.000 mg/mL MTX，即SIN1、2、3、4组；MTX1、2、3、4组以及联合1、2、3、4组。以无药物培养基作为对照组。共13组，每组3个复孔。采用MTT法检测FLS生长情况。采用流式细胞术检测FLS凋亡情况。通过半定量RT-PCR观察FLS核因子κB受体活化因子配体（RANKL）mRNA和骨保护素（OPG）mRNA的表达。
与对照组相比，各药物组RA FLS增殖OD值均降低（P < 0.05）。联合3组RA FLS凋亡OD值升高，OPG mRNA表达升高，RANKL mRNA表达降低，差异有统计学意义（P < 0.05）。SIN3组和MTX3组RA增殖OD值与联合3组相比升高（P < 0.05）。
SIN与MTX在抑制FLS方面具有协同作用。这可能是抑制RA骨破坏的机制之一。
