丹酚酸B对糖氧剥夺损伤大鼠海马神经干细胞增殖、凋亡和分化的影响

Salvianolic acid B effects on the proliferation, differentiation and apoptosis of hippocampal neural stem cells in rats following oxygen-glucose deprivation

背景:丹酚酸B具有改善神经损伤、促进神经发生的作用,但其对海马神经干细胞增殖、凋亡和分化的影响仍不清楚。目的:研究丹酚酸B对体外氧糖剥夺损伤大鼠海马神经干细胞增殖、凋亡及分化的影响。方法:取生长状态良好的新生SD大鼠海马神经干细胞,分6组培养:其中5组均于无糖培养基中在含有体积分数为1%O2、5%CO2和94%N2厌氧混合气体的培养箱中缺氧处理150 min,再分别加入0(对照),5,10,20,40 mg/L的丹酚酸B干预细胞,干预4 d后,MTT检测细胞增殖;干预48 h后,流式细胞术检测海马神经干细胞凋亡;干预5 d后,流式细胞术分析神经元特异性烯醇化酶和胶质纤维酸性蛋白阳性细胞的比例。以正常培养细胞为正常对照。结果与结论:(1)细胞增殖:与正常对照组比较,对照组神经元干细胞增殖能力明显降低(P<0.01);与对照组相比,丹酚酸B处理组神经干细胞增殖能力逐渐升高,以20,40 mg/L丹酚酸B干预组升高明显(P<0.01);(2)细胞凋亡:与正常对照组比较,对照组海马神经干细胞凋亡率明显升高(P<0.01);与对照组比较,丹酚酸B处理组神经干细胞凋亡率随着丹酚酸B质量浓度的增加而逐渐降低,以20,40 mg/L丹酚酸B干预组降低明显(P<0.01);(3)细胞分化:与正常对照组比较,对照组神经干细胞的胶质纤维酸性蛋白阳性细胞比例升高;与对照组比较,不同质量浓度丹酚酸B处理组神经干细胞的神经元特异性烯醇化酶阳性细胞比例升高,胶质纤维酸性蛋白阳性细胞比例下降;(4)结果表明:丹酚酸B可促进氧糖剥夺损伤大鼠海马神经干细胞的增殖,抑制其凋亡,诱导其向神经元分化。

BACKGROUND： Salvianolic acid B can ease nerve injury and promote neurogenesis, but its effects on proliferation,apoptosis and differentiation of neural stem cells in the hippocampus remain unclear.OBJECTIVE： To study the effects of salvianolic acid B on proliferation, apoptosis and differentiation of rat hippocampal neural stem cells following oxygen-glucose deprivation.METHODS： Hippocampal neural stem cells were isolated from newborn Sprague-Dawley rats, and divided into six groups, five of which were cultured in an incubator containing anaerobic mixtures （1% O2, 5% CO2 and 94% N2） for 150minutes followed by treatment with different concentrations of salvianolic acid B （0, 5, 10, 20, 40 mg/L）, respectively.After 4 days of intervention, MTT was used to detect cell proliferation. After 48 hours of intervention, flow cytometry was used to detect cell apoptosis in the hippocampus. After 5 days of culture, flow cytometry was performed to evaluate the percentage of cells positive for neuron-specific enolase and glial fibrillary acidic protein. Normally cultured cells acted as controls （normoxic group）.RESULTS AND CONCLUSION： Compared with the normoxic group, the proliferation of neural stem cells was decreased significantly （P 〈 0.01） and the rate of apoptosis was increased in the oxygen-glucose deprivation group （P 〈0.01）. After treatment with different concentrations of salvianolic acid B, the cell viability and the ratio of neurons in total cells were increased, and the ratio of astrocytes was decreased, especially in 20 and 40 mg/L groups （P 〈 0.01）. In conclusion, these results suggest that salvianolic acid B alleviates adverse effects of oxygen-glucose deprivation on neural stem cell proliferation, differentiation and apoptosis.