丁基苯酞对大鼠骨髓间充质干细胞氧化应激损伤保护机制的研究

Research on the protective mechanism of DL-3-n-butylphthalide on rat bone marrow stem cells under oxidative stress injury induced by hydrogen peroxide

目的研究丁基苯酞(DL-3-n-butylphthalide,NBP)对双氧水(hydrogen peroxide,H2O2)诱导的大鼠骨髓间充质干细胞(rat bone marrow stem cells,rBMSCs)氧化应激损伤的保护机制。方法采用H2O2制作rBMSCs氧化应激损伤模型。实验分为对照组、H2O2损伤组、不同浓度NBP预处理组(0.1μmol组、1μmol组、10μmol组、100μmol组)。不予H2O2及NBP处理的细胞为对照组;H2O2损伤组以终浓度为600μmol的H2O2处理4h制作氧化应激损伤模型;NBP处理组以不同浓度NBP预处理rBMSCs 24h后,再予以终浓度为600μmol的H2O2处理4h。采用MTT法检测各实验组的细胞活力;油镜观察各组细胞形态;流式细胞仪检测各实验组的细胞凋亡率;Westernbolt法检测各实验组Caspase-3的表达情况。结果 NBP可明显降低H2O2对rBMSCs的损伤作用、抑制rBMSCs的凋亡、下调Caspase-3的表达。结论 NBP对H2O2诱导的rBMSCs氧化应激损伤具有保护作用,其机制可能与NBP的抗凋亡作用有关。

Objective To explore the possible protective mechanism of DL-3-n-butylphthalide（NBP） on rat bone marrow stem cells（rBMSCs） under oxidative stress induced by hydrogen peroxide（H2O2）.Methods The oxidative stress of rBMSCs were induced by H2O2.The experiment was carried out on six groups：control group（without the application of H2O2 or NBP）,model group（cells were administrated by H2O2 without NBP pretreatment）,NBP pretreated groups（cells were pretreated with different doses of NBP as 0.1μmol,1μmol,10μmol and 100μmol for 24h,then administrated with 600 μmol H2O2 for 4h）.MTT analysis was used to detect the cell viability of all the groups.The rate of cell apoptosis was observed by flow cytometry analysis.The expression level of protein Caspase-3 was detected by Western blot.Results Compared with model group,MTT and cytometrical analysis showed that pretreatment with NBP could protect rBMSCs from oxidative stress dose-dependently.Up-regulation of protein Caspase-3 induced by H2O2 was significantly reduced by NBP pretreatment.Conclusion NBP has protective effects on rBMSCs challenged by H2O2.The protective mechanism may be related with the antiapoptotic effects provided by NBP.