丹参酮ⅡA促进氧糖剥夺再灌注神经干细胞的增殖作用

Tanshinone ⅡA Promotes the Proliferation of Primary Neural Stem Cells under Oxygen and Glucose Deprivation Reperfusion Conditions

目的观察丹参酮ⅡA对原代培养的小鼠神经干细胞(NSCs)在氧糖剥夺再灌注(OGD/R)损伤条件下增殖作用的影响。方法对原代小鼠NSCs进行培养、鉴定;采用CCK-8法测定细胞活力;通过检测神经球细胞直径来观察NSCs的增殖情况;对OGD/R损伤条件进行筛选,建立OGD/R NSCs模型。结果与溶剂对照组相比,0.03、0.1、0.3、1、3、10、30μmol·L^(-1)丹参酮ⅡA干预48 h后,NSCs的细胞活力显著提高(P<0.01);0.3、1、3μmol·L^(-1)丹参酮ⅡA干预48 h后,NSCs神经球细胞透光性良好,神经球细胞直径明显增大(P<0.05,P<0.01)。NSCs糖氧剥夺4 h再灌注后的细胞活力介于40%~60%之间,故选择OGD/R(4 h/20 h)作为NSCs的OGD/R损伤条件。与溶剂对照组比较,模型组NSCs神经球细胞直径明显缩小、细胞活力明显下降(P<0.01);与模型组比较,3μmol·L^(-1)丹参酮ⅡA干预48 h后,NSCs神经球细胞直径明显增大、细胞活力明显升高(P<0.01)。结论丹参酮ⅡA可促进正常条件及OGD/R损伤条件下NSCs的增殖及其细胞活力,丹参酮ⅡA对脑缺血损伤的保护作用机制值得进一步探讨。

Objective To observe the effect of tanshinoneⅡA on the proliferation of mouse primary neural stem cells(NSCs) after oxygen and glucose deprivation reperfusion(OGD/R) injury.Methods Primary NSCs of mouse were cultured and identified.Cell viability was determined by CCK-8 assay.The proliferation of NSCs was observed by measuring the diameters of nerve bulb cells.The OGD/R injury conditions were screened and the OGD/R NSCs model was established.Results Compared with the solvent control group,NSCs cell viability was significantly increased after treatment with 0.03,0.1,0.3,1,3,10,30μmol·L^(-1)tanshinoneⅡA for 48 h(P<0.01).After0.3,1,3μmol·L^(-1)tanshinoneⅡA treatment for 48 h,NSCs nerve bulb cells showed good light transmittance and the diameters of nerve bulb cells increased significantly(P<0.05,P<0.01).Viabilities of the NSCs with reperfusion after 4 h glucose and oxygen deprivation ranged from 40%to 60%;and OGD/R(4 h/20 h)was selected as the OGD/R condition of NSCs.Compared with solvent control group,the diameter and cell viability of NSCs in model group were significantly reduced(P<0.01).Compared with model group,diameters of NSCs neuronal bulb cells and the cell viability were significantly increased after 3μmol·L^(-1)tanshinoneⅡA treatment for 48 h(P<0.01).Conclusion TanshinoneⅡA can promote the proliferation and cell viability of NSCs under normal condition and OGD/R injury,and the mechanism of tanshinoneⅡA on cerebral ischemia injury deserves is worthy of further study.