钙离子一氧化氮在硒化壳聚糖诱导的皮肤成纤维细胞增殖中的作用  

Effects of intracellular calcium and nitric oxide on proliferation of cultured human skin fibroblasts induced by selenium chitosan in vitro

作　　者：邓守恒[1] 张军[1] 姜立东[1] 许涛[1] 李芳[1] 陈萍[1] 

机构地区：[1]郧阳医学院附属人民医院,442000

出　　处：《山西医药杂志（上半月）》2010年第5期412-414,共3页Shanxi Medical Journal

基　　金：郧阳医学院附属太和医院科研项目(ky20031207)

摘　　要：目的观察硒化壳聚糖对体外培养人皮肤成纤维细胞增殖周期及细胞内一氧化氮(NO)水平及钙离子含量的影响。方法皮肤成纤维细胞培养液加入不同浓度的硒化壳聚糖(25、50、100、200、400 mg/L),应用噻唑蓝(MTT)法检测细胞增殖,改良Griess法检测NO,Huo-3AM钙荧光指标剂测定细胞内钙离子浓度,流式细胞仪检测细胞周期。结果与对照组比较,50~400 mg/L硒化壳聚糖作用48 h后,细胞对MTT的吸收明显升高(P<0.05);各浓度硒化壳聚糖组细胞NO水平(72±9)μmol/L^(44±10)μmol/L与对照组(107±9)μmol/L比较均呈不同程度下降,呈剂量依赖性(P<0.01);200、400 mg/L硒化壳聚糖组细胞内钙离子浓度(179±32)mmol/L、(179±14)mmol/L与对照组(147±10)mmol/L比较显著升高(P<0.05);流式细胞分析显示,G0~G1期细胞所占百分率显著性减少,G2~M期和S期细胞所占百分率显著性增加(P<0.01)。结论硒化壳聚糖可通过调节细胞内钙离子含量和NO水平来促进皮肤成纤维细胞增殖。Objective To investigate the effects of intracellular calcium and nitric oxide on proliferation of cultured human skin fibroblasts induced by selenium chitosan in vitro.Methods The cultured human skin fibroblasts were treated with 25,50,100,200 and 400 mg/L of selenium chitosan.The proliferation of the cell was measured by MTT analysis.NO level was assayed by Griess method.The concentration of intracellular calcium was determined using the fluorescent calcium indicator Fluo-3Am and the cell cycle was analyzed by flow cytometer.Results The absorption to MTT of the cells treated with 50 mg/L to 400mg/L of selenium chitosan for 24 hours increased dramatically compared with that of control group（P〈0.05）.The level of NO was markedly decreased in a dose-dependent manner（〈P0.01） in cells treated with 25 mg/L to 400 mg/L of selenium chitosan for 48 hours.Compared with control group,the concentration of intracellular calcium was significantly increased in 200 mg/L and 400 mg/L selenium chitosan group（P〈0.05）.The flow cytometer analysis of the cell cycle distribution also revealed that the percentage of phase G0-G1 was markedly decreased,and G2~M and S was significantly increased（P〈0.01） in selenium chitosan group.Conclusion Selenium chitosan could promote the proliferation of the human skin fibroblasts by regulating intracellular calcium concentration and inhibiting the release of NO.

关 键 词：成纤维细胞 壳聚糖 增殖 钙离子 NO 

分 类 号：R751[医药卫生—皮肤病学与性病学]