10-羟基喜树碱对类风湿关节炎滑膜成纤维细胞的增殖抑制作用  

Effect of 10-Hydroxycamptotbecine on the proliferation of human Fibroblast-like Synoviocyte with Rheumatoid Arthritis

作　　者：任丽丽[1] 鲁晓娟 寿记新[1] 王燕[1] 孙晓麟[2] 张丽娟[1] 张濛[1] 孟洋[1] 孙贺[1] 

机构地区：[1]郑州大学第五附属医院,450002 [2]北京大学人民医院风湿免疫科 [3]郑州铁路职业技术学校

出　　处：《中华医学杂志》2016年第42期3389-3392,共4页National Medical Journal of China

基　　金：河南省教育厅重点科技攻关项目（14A320053）

摘　　要：目的探讨10-羟基喜树碱（10-HCPT）对类风湿关节炎（RA）患者滑膜成纤维细胞（FLS）的增殖抑制作用。方法用不同剂量的10-HCPT和甲氨蝶呤（MTX）处理类风湿关节炎和骨关节炎（OA）滑膜成纤维细胞，采用CCK-8法检测细胞增殖抑制，Annexin-VAPC／7-AAD染色法检测细胞凋亡。结果经1．0mg／L、10．0mg／L10．HCPT组作用48h和72h后的RA滑膜成纤维细胞的存活率分别为（66．68±0．48）％与（44．05±1．29）％、（60．09±0．95）％与（30．63±1．79）％。而MTX组48h和72h后的RA滑膜成纤维细胞的存活率分别为（78．24±0．99）％与（72．80±1．39）％、（69．08±0．86）％与（56．79±2．33）％。作用过的RA及OA滑膜成纤维细胞存活率均下降，且随着作用时间的延长，细胞存活率进一步下降。与MTX组相比10．0mg／L10-HCPT组可明显抑制滑膜成纤维细胞的增殖（P〈0．05）。经1．0mg／L、10．0mg／L10-HCPT组作用48、72h后的RA滑膜成纤维细胞的凋亡率分别为（24．87±2．69）％与（35．45±3．79）％、（30．10±2．97）％与（46．16±0．99）％。与对照组比较（16．38±1．68）％，两种浓度的10-HCPT、MTX[（20．46±4．87）％与（25．37±0．98）％、（22．14±1．86）％与（28．26±3．95）％]均可诱导RA及OA滑膜成纤维细胞的凋亡，其凋亡率高于对照组；随着作用时间的延长（72h），诱导凋亡的作用显著增强（P〈0．05）。对于RA滑膜成纤维细胞，作用时间为48h，10-HCPT组诱导滑膜成纤维细胞的凋亡率高于相同浓度的MTX组，其中10．0mg／L10．HCPT组诱导凋亡作用更强（P〈0．05）；作用72h时，10．0mg／L10．HCPT组凋亡率明显高于其他各组（P〈0．05）。结论10-HCPT具有较MTX更强的抑制细胞增殖、促进细胞凋亡的作用。Objective To study the effect of 10-Hydroxycamptothecine （10-HCPT） on the proliferation and apoptosis of human Fibroblast-like Synoviocyte （FLS） with Rheumatoid Arthritis （RA）. Methods Different concentrations of 10-HCPT and Methotrexate （MTX） were used to treat FLS cells in RA and Osteoarthritis （OA） for different time （24, 48, and 72 hours）, and FLS cells without 10-HCPT and MTX were served as the control group. CCK-8 assay were applied to determine the proliferation of FLS cells, Aunexin-V APC/7-AAD staining were used to detect the apoptosis of FLS cells. Results The survival rate of FLS cells were （66.68±0.48） %,48h; （60.09±0.95） %, 72 h and （44.05 ±1.29） %, 48 h; （30.63 ± 1.79） %, 72 h, when the concentrations were 1.0 μg/ml and 10.0 μg/ml in 10-HCPT group. Compared with the control group, the survival rate of FLS cells in RA and OA both declined in treatment groups with different concentrations of 10-HCPT and MTX. With the extension of time, the survival rate of FLS cells declined significantly. Compared with the MTX group, there were no obvious differences in 10- HCPT group with 1.0 μg/ml. But the concentration of 10.0 μg/ml of 10-HCPT group showed obviously difference in the proliferation of FLS cells. The apoptosis rate of FLS cells were （66. 68 ± 0. 48） %, 48 h ; （60.09±0.95） %, 72 h and （44.05 ± 1.29） %, 48 h; （30.63 ± 1.79） %, 72 h, when the concentrations were 1.0 μg/ml and 10. 0 μg/ml in 10-HCPT group. Compared with the control group, two concentrations of 10-HCPT and MTX induced higher apoptosis in FLS cells with RA and OA; with the extension of time （72 h）, the rate of apoptosis was significantly enhanced （ P 〈 0.05 ）. When FLS cells with RA were treated for 48 h, apoptosis of 10-HCPT group was higher than that of MTX group. The 10. 0 μg/ml of 10-HCPT had the highest effect. Conclusion Compared with MTX, 10-HCPT had the higher efficacy of inhibiting proliferation and promoting apoptosis in FLS cells.

关 键 词：10-羟基喜树碱 类风湿关节炎 滑膜成纤维细胞 凋亡 增殖 

分 类 号：R593.22[医药卫生—内科学]