青藤碱对类风湿关节炎患者成纤维样滑膜细胞体外增殖及凋亡的影响

Effects of sinomenine on in vitro proliferation and apoptosis of fibroblast-like synoviocyte from patients with rheumatoid arthritis

目的观察青藤碱(sinomenine,SN)对类风湿关节炎(rheumatoid arthritis,RA)患者成纤维样滑膜细胞(fi-broblast-like synoviocytes,FLS)凋亡及增殖的影响,为青藤碱在临床上的应用提供实验依据。方法光镜和荧光显微镜观察FLS形态的变化,流式细胞仪检测细胞凋亡率,MTT比色法检测SN对FLS生长的抑制作用。结果光镜及荧光显微镜下可见核固缩,变圆,荧光染色增强等凋亡形态学变化。流式细胞仪结果显示,随着SN浓度及作用时间的增加,被处理的FLS的凋亡率增加,未处理组与各处理组比较差异具有显著性(P<0·05),其中3.2mmol/L浓度的SN作用48h对FLS凋亡影响最为显著。MTT结果显示,加入不同浓度SN(2.8～3.2mmol/L),培养的FLS24h、48h后对增殖的抑制水平显著高于未处理组,与凋亡率呈现正相关(r=0.787,P<0·05)。结论SN在一定浓度及作用时间内可抑制RA患者FLS增殖,诱导凋亡,这可能是SN治疗RA机制之一。

Objective To observe the effect of sinomenine （SN） on in vitro apoptosis and proliferation of fibroblast-like synoviocytes （FLS） from patients with rheumatoid arthritis. Methods The morphological changes of FLS were observed by invert microscope and fluorescence microscope. The amount of apoptotic cells was measured by flow cytometry. The inhibition rate of FLS was examined by MTY assay. Results The morphological changes of apoptosis, such as cell rounding, shrinkage and enhanced fluorescein stain, were observed. Flow cytometry demonstrated that the apoptosis rate increased with increased SN concentration and treatment time. The difference of apoptosis rate between the treated and non-treated group was significant （ P 〈 0. 05 ）. The effect of SN on the FLS apoptosis was most obvious when SN was 3.2 mmol/L and the treatment time was 48 h. The cell inhibition of SN-treated group （2.8, 3. 0, 3.2 mmol/L） was significantly increased as compared with corresponding non-treated group （P 〈0. 01 ） by MTT assay. The inhibition rate of FLS was found positively correlated with apoptosis rate under the treatment of SN （P 〈 0. 05）. Conclusion In a certain concentration and treatment time, SN can inhibit FLS proliferation and induce apoptosis. This may be one of the therapeutic mechanisms of SN on RA.