摘要
目的研究不同浓度沙利度胺对体外培养成纤维细胞样滑膜细胞(FLS)凋亡的影响。方法体外培养及纯化FLS,分为6组:对照组,脂多糖(LPS)组,LPS+高剂量沙利度胺(50mg/L)组,LPS+中剂量沙利度胺(5mg/L)组,LPS+低剂量沙利度胺(0.5mg/L)组,LPS+甲氨蝶呤(MTX)组。48小时后显微镜及电镜观察不同处理组FLS形态,膜黏连蛋白-5/碘化丙啶(Annexin-Ⅴ/PI)联合标记法测定FLS凋亡率及坏死率。结果①原代滑膜细胞培养3代后以梭形且呈极向排列FLS为主;②沙利度胺及MTX处理后FLS渐呈不规则形,细胞变圆缩小,透射电镜下可见典型细胞凋亡。③中、高剂量沙利度胺及MTX组凋亡率和坏死率均高于对照组、LPS组及低剂量沙利度胺组,凋亡率分别为(5.72±1.42)%、(8.07±1.45)%、(11.28±2.48)%vs(0.30±0.09)%、(0.24±0.05)%、(0.40±0.04)%(P<0.05),坏死率分别为(12.96±4.32)%、(20.34±4.43)%、(18.11±1.17)%vs(0.22±0.08)%、(0.13±0.09)%、(1.94±1.16)%(P<0.05);低、中、高剂量沙利度胺及MTX组凋亡率均依次升高(P<0.05);低、中、高剂量沙利度胺组及MTX组坏死率均依次升高(P<0.05);高剂量沙利度胺组与MTX组坏死率之间比较差异无统计学意义(P>0.05)。结论沙利度胺能够诱导FLS凋亡,在一定浓度范围内该作用呈剂量依赖性。沙利度胺对FLS早期凋亡作用弱于MTX,对晚期凋亡作用与MTX相同。
Objective To investigate the effect of thalidomide in different dosages on apoptosis of fibroblast-like synoviocytes（FLS） cultured in vitro in the experimental arthritis rats.Methods The FLS were successfully cultured and purified in vitro from the synovial membrane tissues of the experimental arthritis rats,and the cells were divided into six groups as following：control group,lipopolysaccharide（LPS） group,LPS with high dosage of thalidomide（50 mg/L） group,LPS with middle dosage of thalidomide（5 mg/L） group,LPS with low dosage of thalidomide（0.5 mg/L） group and methotrexate（MTX） group.The morphological characteristics of FLS were observed by convert microscope and transmission electron microscope,and the apoptosis and necrosis rates were determined by flow cytometry after 48 hours.Results ①The original synovial cells tended to be spindle-shaped cells with good cell homogeneity after the third passage;② FLS changed to anomalistic form with smaller size,and cell apoptosis was distinct by the transmission electron microscope;③ The apoptosis and necrosis rates of the middle and high dosage of thalidomide group and MTX group were all higher than those of control group,LPS group and low dosage of thalidomide group（P0.05）.The apoptosis rates were（5.72±1.42）%,（8.07±1.45）%,（11.28±2.48）% vs（0.30±0.09）%,（0.24±0.05）%,（0.40±0.04）%（P0.05） and the necrosis rates were（12.96±4.32）%,（20.34±4.43）%,（18.11±1.17）% vs（0.22±0.08）%,（0.13±0.09）%,（1.94±1.16）%（P0.05）,respectively.The apoptosis rates of the low,middle and high dosage of thalidomide group and MTX group increased in turn（P0.05） and the same results were verified in the necrosis rates of the low,middle and high dosage of thalidomide group.Compared with MTX group,there was no difference in the necrosis rates in high dosage of thalidomide group（P0.05）.Conclusion Thalidomide could induce FLS apoptosis partly in a dose-dependent manner,and it functions the same as MTX in late apoptosis but not better in earlier apoptosis than MTX.
出处
《临床荟萃》
CAS
2012年第10期865-868,F0003,共5页
Clinical Focus
关键词
关节炎
实验性
滑膜
细胞凋亡
沙利度胺
arthritis
experimental
synovial membrane
apoptosis
thalidomide
