葡萄糖转运蛋白1抑制剂BAY-876对类风湿性关节炎滑膜细胞增殖及转移的抑制作用  

Inhibitory effect of glucose transporter 1 inhibitor BAY-876 on the proliferation and metastasis of human synovial cells from rheumatoid arthritis

作　　者：甄亚男 甄诚 葛贤明 郭滨 魏芳 韦颖梅 刘浩 ZHEN Yanan;ZHEN Cheng;GE Xianming;GUO Bin;WEI Fang;WEI Yingmei;LIU Hao(School of Pharmacy,Bengbu Medical College,Anhui Provincial Engineering Technology Research Center of Biochemical Pharmaceuticals,Benbu 233030,China)

机构地区：[1]蚌埠医学院药学院安徽省生化药物工程技术研究中心

出　　处：《山西医科大学学报》2019年第9期1272-1277,共6页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(81372899,81703529);安徽省对外科技合作项目(1503062024);安徽高校自然科学研究重点项目(KJ2018A0231);安徽省自然科学基金项目(1508085MH166)

摘　　要：目的 探讨葡萄糖转运蛋白1抑制剂BAY-876对类风湿性关节炎(rheumatoid arthritis,RA)成纤维样滑膜细胞(fibroblast-like synoviocytes,FLS)增殖、迁移及侵袭能力的抑制作用。方法 不同浓度的BAY-876(0,0.5,1,2,4,8 μmol/L)处理RA-FLS细胞24,48,72 h,CCK-8法检测BAY-876对细胞的增殖抑制作用。ATP试剂盒检测不同浓度BAY-876(0,2,4,8 μmol/L)处理RA-FLS细胞24 h后,细胞内ATP水平的变化;Transwell法和细胞划痕实验检测不同浓度BAY-876(0,2,4,8 μmol/L)刺激下RA-FLS细胞的迁移与侵袭能力的改变;Western blot法检测细胞中MMP-2、MMP-9及AKT蛋白的表达。结果 CCK-8法实验结果表明,与对照组相比,不同浓度(0.5,1,2,4,8 μmol/L)的BAY-876作用24 h后的细胞存活率依次为(90.9 ± 2.9)%、(85.6 ± 5.4)%、(82.7 ± 7.6)%、(72.4 ± 1.9)%、(45.0 ± 0.7)%。且随着药物浓度的增加和处理时间延长,细胞存活率下降( P <0.05)。ATP水平检测结果表明,2,4,8 μmol/L BAY-876可明显降低RA-FLS细胞内的ATP水平( P <0.01)。Transwell法检测不同浓度(2,4,8 μmol/L)BAY-876处理细胞后,穿过小室膜的细胞数明显少于对照组;迁移试验表明,2,4,8 μmol/L BAY-876药物组的迁移抑制率分别为(16.2 ± 3.9)%、(60.1 ± 3.0)%和(68.2 ± 2.3)%,侵袭抑制率分别为(62.6 ± 3.3)%、(85.3 ± 3.7)%和(91.8 ± 3.2)%,与对照组比较差异均具有统计学意义( P <0.01)。Western blot结果显示,BAY-876可下调MMP-2、MMP-9和AKT的表达。结论 葡萄糖转运蛋白1抑制剂BAY-876可以显著地抑制对体外培养RA FLS细胞的增殖、迁移及侵袭能力,下调基质金属蛋白酶的表达,可以作为潜在的治疗风湿性关节炎的药物。Objective To investigate the effects of glucose transporter 1 inhibitor BAY-876 on the proliferation, migration and invasion of fibroblast-like synoviocytes(FLS) from rheumatoid arthritis(RA). Methods RA-FLS were treated with different concentrations of BAY-876(0,0.5,1,2,4,8 μmol/L) for 24,48,72 h, and then the cell proliferation was detected by CCK-8 assay. RA-FLS were treat-ed with different concentrations(0,2,4,8 μmol/L) of BAY-876 for 24 h, and then the intracellular adenosine triphosphate(ATP) concentration was measured by the commercial ATP detection kit, the cell invasion and migration were examined by Transwell assay and wound healing assay, and the expression of MMP-2, MMP-9 and AKT was analyzed by Western blot. Results CCK-8 assay showed that compared to control group(0 μmol/L), BAY-876 effectively decreased the cell viability of RA-FLS in time-dependent and concentration-dependent manners( P <0.05), and the cell viability was (90.9 ± 2.9)%,(85.6 ± 5.4)%,(82.7 ± 7.6)%,(72.4 ± 1.9)%,(45.0 ± 0.7)% after treated with different concentrations(0.5, 1, 2, 4, 8 μmol/L) at 24 h, respectively. Furthermore, 2, 4, 8 μmol/L BAY-876 significantly decreased the intracellular ATP level( P <0.01). Compared with control group(0 μmol/L), 2, 4, 8 μmol/L BAY-876 significantly inhibited cell migration and invasion of RA-FLS( P <0.01), and the inhibition rates of migration were (16.2 ± 3.9)%,(60.1 ± 3.0)% and (68.2 ± 2.3)%, respectively, and the inhibition rates of invasion were (62.6 ± 3.3)%,(85.3 ± 3.7)% and (91.8 ± 3.2)%, respectively. In addition, Western blot analysis revealed that BAY-876 effectively reduced the expression of MMP-2, MMP-9 and AKT in RA-FLS. Conclusion BAY-876 can significantly inhibit the proliferation, invasion and migration of RA FLS, and down-regulate expression of matrix metalloproteinase, which can be a potential drug for rheumatoid arthritis.

关 键 词：类风湿性关节炎 葡萄糖转运蛋白1 BAY-876 增殖 迁移 侵袭 

分 类 号：R593.22[医药卫生—内科学]