海藻多糖对类风湿关节炎成纤维样滑膜细胞凋亡的影响及机制研究

作者	单位
朱浪静	中山大学孙逸仙纪念医院风湿免疫科
郑东辉	中山大学孙逸仙纪念医院风湿免疫科
张白玉	中山大学孙逸仙纪念医院风湿免疫科
闻克威	中国科学院南海海洋研究所
莫颖倩	中山大学孙逸仙纪念医院风湿免疫科
李婷	广州医学院第三附属医院血液风湿科
高璇	中国科学院南海海洋研究所
戴冽	中山大学孙逸仙纪念医院风湿免疫科

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中文摘要:

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。

英文摘要:

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(P<0.01) 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(P<0.05,P<0.01).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.

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