摘要
目的探讨海藻多糖对体外培养的类风湿关节炎(rheumatoid arthritis,RA)成纤维样滑膜细胞(fibroblast-like synovialcytes,FLS)凋亡的影响及机制。方法采用改良组织块培养法体外培养RA-FLS。分别采用CCK-8比色法、Hoechst 33258染色法、TUNEL染色法及Western blot检测不同浓度海藻多糖(0、15、20、25mg/mL)干预不同时间(0、3、4、5天)后对RA-FLS增殖、凋亡及Caspase-3、Bax、Bcl-2蛋白表达的影响。结果与0mg/mL海藻多糖组同期比较,15、20、25mg/mL浓度组干预3、4、5天后均可抑制RA-FLS增殖,促进其凋亡(P<0·01),且具有时间和剂量依赖性;15、20、25mg/mL海藻多糖干预4天可上调RA-FLSBax蛋白表达,下调Bcl-2蛋白表达,促进Caspase-3活化裂解(P<0·05,P<0·01),且呈剂量依赖性。结论海藻多糖在体外呈剂量和时间依赖性抑制RA-FLS增殖,诱导其凋亡,其凋亡机制可能是通过上调细胞内Bax蛋白及下调Bcl-2蛋白表达影响线粒体通路,从而促进Caspase-3活化裂解,导致RA-FLS凋亡。
Objective To study the effect and related mechanism of seaweed polysaccharide（SP） on apoptosis of fibroblast-like synoviocytes in patients with rheumatoid arthritis（RA-FLS）.Methods RA-FLS were in vitro cultured using modified tissue culture method.Effect of SP（0,15,20,and 25 mg/mL,respectively） at different time points（0,3,4,and 5 days,respectively） on the proliferation and apoptosis of RA-FLS,and protein expressions of Caspase-3,Bax,and Bcl-2 was detected by cell counting kit-8（CCK-8） assay,Hoechst 33258 staining assay,TUNEL assay,and Western blot,respectively.Results Compared with 0 mg/mL SP at the same time point,the proliferation of RA-FLS was inhibited,and the apoptosis was promoted 3,4,and 5 days after intervened by 15,20,and 25 mg/mL SP,respectively（P0.01） in time-and dose-dependent manners.RA-FLS Bax protein expression was up-regulated,Bcl-2 protein expression down-regulated,Caspase-3 activated and split by 15,20,and 25 mg/mL SP,respectively for 4 days（P0.05,P0.01）.Besides,the changes were in a dose-dependent manner.Conclusions SP could inhibit RA-FLS proliferation and induce its apoptosis in dose-and time-dependent manners.Its apoptosis mechanism might be through up-regulating intracellular Bax protein expression and down-regulating Bcl-2 protein expression,thus influencing the mitochondrion signaling pathway,further promoting Caspase-3 activation and split,resulting in the apoptosis of RA-FLS.
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2011年第7期961-966,共6页
Chinese Journal of Integrated Traditional and Western Medicine
基金
国家自然科学基金面上项目(No.30972742)
广东省科技计划项目(No.2008B030301093)
中国科学院海洋生物资源可持续利用重点实验室(LMB)开放基金(No.LMB071010)
关键词
类风湿关节炎
成纤维样滑膜细胞
海藻多糖
凋亡
rheumatoid arthritis
fibroblast-like synoviocyte
seaweed polysaccharide
apoptosis
