三氧化二砷对高迁移率族蛋白诱导的大鼠滑膜细胞增殖的抑制作用

目的： 探讨三氧化二砷（AS₂O₃）对高迁移率族蛋白1（HMGB1）诱导的大鼠滑膜细胞(RSC-364细胞)增殖的抑制作用及其机制。 方法： 将常规培养的RSC-364细胞分为正常对照组、HMGB1组及HMGB1加0、5、10、20、40和80 μmol?L^-1 AS₂O₃实验组； MTT法检测不同浓度AS₂O₃对RSC-364细胞生长的影响，RT-PCR检测STAT1 mRNA的表达变化，免疫细胞化学和流式细胞术检测STAT1、cyclin D1和增殖细胞核抗原（PCNA）蛋白表达的变化。结果：① MTT结果显示，AS₂O₃可抑制HMGB1诱导的RSC-364细胞的增殖，80　μmol?L^-1 AS₂O₃作用24　h抑制率可达88.18%，其抑制作用呈时间和浓度依赖性（P<0.05或P<0.01）；② 与对照组比较，HMGB1组STAT1 mRNA及STAT1、PCNA 、cyclin D1蛋白表达量增强，AS₂O₃组STAT1 mRNA及STAT1、PCNA、cyclin D1蛋白降低，并呈剂量依赖性(P<0.01）；③ STAT1与cyclin D1蛋白表达呈正相关关系（r=0.842，P=0.001）。结论：AS₂O₃能够抑制HMGB1诱导的滑膜细胞的增殖，其机制可能与抑制STAT1的活性从而下调细胞周期调节蛋白cyclin D1的表达有关。

Inhibitory effect of arsenic trioxide on proliferation of rat synoviocyte stimulated by HMGB1

Objective To investigate the inhibitory effect of arsenic trioxid(AS2O3)on the proliferation of RSC-364 synoviocyte lines stimulated by HMGB1 and its possible mechanism. Methods RSC-364 synoviocytes were cultivated with standard medium as control group or with medium supplemented with HMGB1(HMGB1 group) or HMGB1 and 0,5,10,20,40,80 μmol·L-1 AS2O3(experimental groups),respectively. MTT assay was carried out to study the inhibitory effects of AS2O3 with different concentrations on the growth of RSC-364. RT-PCR was used to detect the mRNA expression of STAT1. Proteins of phospho-STAT1,cyclin D1 and proliferation cell nuclear antigen (PCNA) were detected by immunocytochemistry and FCM.Results ① MTT data indicated AS2O3 could inhibit the growth of RSC-364 stimulated by HMGB1 in a time-dependent and dose-dependent manner. ②Compared with control group,HMGB1 up-regulated the expression of phospho-STAT1 protein and mRNA as well as PCNA protein. Both mRNA and protein levels of phospho-STAT1 were decreased in the RSC-364 cells exposed to AS2O3 in a dose-dependent manner; AS2O3 also down-regulated the expression of PCNA protein. ③ HMGB1 up-regulated the expression of cyclin D1,while AS2O3 decreased the expression of cyclin D1 .④ There was positive correlation between phospho-STAT1 and cyclin D1 (r=0.842,P=0.001);.Conclusion Arsenic trioxide could inhibit the proliferation of RSC-364 synoviocytes,partly by down-regulating the expression of phospho-STAT1/cyclin D1 .