甲氨蝶呤诱导色素沉着绒毛结节性滑膜炎滑膜细胞凋亡的研究

The Effect of MTX on Synoviocytes Apoptosis in Pigmented Villonodular Synovitis

目的:观察甲氨蝶呤(Methotrexate,MTX)对色素沉着绒毛结节性滑膜炎(pigmented villonodular synovitis,PVNS)滑膜细胞增殖和凋亡的影响。方法:实验分成不同浓度的MTX药物处理组,以未加药(MTX浓度为0mmol/L)组为空白对照组。酶消化法进行人PVNS关节滑膜原代分离培养,体外培养的PVNS滑膜细胞空白对照组和MTX药物处理组加药后分别于48、96、144h用MTT比色法检测MTX对PVNS滑膜细胞增殖的抑制率,流式细胞仪检测MTX处理48h后对细胞凋亡的影响。结果:与空白对照组相比,不同浓度的MTX均显著抑制PVNS滑膜细胞的增殖,差异有统计学意义(P<0.05),MTX体外作用PVNS滑膜细胞48h后,空白对照组和0.5、1.0、2.0mmol/L MTX药物处理组细胞凋亡率分别为(5.589±0.852)%、(6.857±0.604)%、(8.787±0.196)%和(11.037±0.729)%,1.0、2.0mmol/L MTX药物处理组细胞凋亡率与空白对照组比较,差异均有统计学意义(P<0.05)。结论:MTX可抑制体外培养的PVNS滑膜细胞的增殖活力,并可诱导滑膜细胞的凋亡。

Objective： To study the effect of MTX on the proliferation and apoptosis of synoviocyteson synovial cells in human pigmented villonodular synovitis. MethOdS： The experiment included three concentrations of treatment groups （MTX） and control group. Primary synovial ceils were cultured by means of enzymatic digestion from PVNS patient. The inhibition of synoviocytes viability was detected by MTr method after the cells treated with MTX for 48 hours, 96 hours and 144 hours. Apoptosis changes induced by MTX were examined by flow cytometry after treated for 48 hours. Results： MTX could obviously inhibit the proliferation of PVNS synovial cells compared with the control group （P 〈 0.05）. The apoptosis were ob- served after 48 hours. Apoptosis rates of control group and MTX 0.5 mmol/L, 1.0 mmol/L, 2.0 mmol/L were （5.589± 0.852）%, （6.857±0.604）%, （8.787±0.196）%, and （11.037±0.729）% respectively. There was a significant difference in apoptosis rate between 1.0 and 2.0 mmol/L MTX groups （P 〈 0.05）. Cell apoptosis rates in MTX groups were higher than that in control group. Conclusion： MTX can inhibit the proliferation and induce the apoptosis of synoviocytes in PVNS. Key words Methotrexate synovitis, pigmented villonodular cell proliferation apoptosis