氧糖剥夺/复氧损伤后星形胶质细胞iNOS的表达及胍丁胺的干预作用研究

Effect of agmatine on iNOS expression after ischemia-reperfusion injury in astrocytes

目的观察星形胶质细胞缺血再灌注损伤后iNOS的表达及胍丁胺的干预作用。方法体外缺血再灌注诱导损伤原代培养的星形胶质细胞,检测培养液中乳酸脱氢酶(lactate dehydrogenase,LDH)漏出量,并利用流式细胞术检测细胞凋亡和/或坏死率,同时测定NO生成量和诱导型一氧化氮合酶(inducible NOS,iNOS)的表达。结果胍丁胺显著提高缺血再灌注损伤细胞的存活率,降低细胞LDH漏出量。流式细胞仪分析显示胍丁胺主要是降低细胞坏死率而对凋亡率则无显著性影响。与阳性对照组相比,胍丁胺显著降低星形胶质细胞缺血再灌注损伤引起的NO产量,同时也减少了iNOS表达。结论胍丁胺对星形胶质细胞缺血再灌注损伤具有保护作用,其可能的作用机制是降低iNOS、抑制炎症因子NO合成,从而降低星形胶质细胞的炎症坏死。

Objective To investigate the protective effect and related mechanisms of agmatine on murine brain astrocytes after ischemia-reperfusion injury.Methods The astrocytes were subjected to ischemia-reperfusion injury （oxygeon glucose deprivation-reperfusion,OGD-R）.The efflux of lactate dehydrogenase （LDH） and flow cytometric analysis were determined. Also, the production of NO were measured,the expression of inducible NOS （iNOS） were detected by western blot analysis.Results The viability of astrocyte was increased and the efflux of LDH were reduced by agmatine treatmented.By FITC-Annexin V/PI staining, the data demonstrated that agmatine decreased the necrosis rate but not the apotosis rate.The contents of NO production were reduced,and the expression of iNOS were decreased by agmatine, too.Conclusion Agmatine has a protective effect on astrocyte after ischemia-reperfusion injury,which maybe related to its inhibitory effect on iNOS-NO pathway.