远志皂苷元对氧糖剥夺/再复氧诱导大鼠海马神经干细胞损伤的保护作用
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摘要
目的:探讨远志皂苷元对新生大鼠海马神经干细胞(NSCs)氧糖剥夺/再复氧(OGD/R)诱导损伤的保护作用。方法:体外培养新生大鼠海马NSCs,将细胞分为对照组、OGD/R组和远志皂苷元组。OGD/R组用无糖Earle’s液于缺氧后正常培养,远志皂苷元组在OGD/R基础上加远志皂苷元干预培养。MTT法测定NSCs细胞存活率,比色法检测乳酸脱氢酶(LDH),DAPI染色检测NSCs细胞的凋亡,Nestin/BrdU双标染色检测NSCs细胞的增殖。结果:与对照组比较,OGD/R组NSCs的OD值、存活率、Nestin/BrdU阳性反应显著降低(P<0.01),LDH漏出率显著增加(P<0.01),DAPI核染提示OGD可诱导细胞凋亡。与OGD/R组比较,远志皂苷元组NSCs的OD值、存活率、Nestin/BrdU阳性反应的面密度及细胞数显著增多(P<0.01),LDH漏出率显著减少(P<0.01),DAPI核染显示远志皂苷元可抑制细胞凋亡。结论:远志皂苷元对OGD/R诱导大鼠海马NSCs的损伤具有改善作用。
Objective: To explore the protective effect of senegenin on the injury of rat hippocampal neural stem cells(NSCs) induced by oxygen-glucose deprivation/reoxygenation(OGD/R) culture in vitro. Methods: Neonatal rat hippocampal NSCs cultured in vitro were randomly divided into control group, OGD/R group, and senegenin group. OGD/R group was cultured by Earle's without glucose in normal culture after hypoxia. Senegenin group was curtured on the basis of OGD/R combined with senegenin. The MTT method was used to detect cell survival rate, the colorimetry was used to detect lactate dehydrogenase(LDH),the DAPI staining method was used to detect apoptosis of NSCs cells, and proliferation of NSCs cells was detected by doublelabeling immunofluorescence method of Nestin/BrdU. Results: Compared with the control group, NSCs OD value, cell survival rate, and Nestin/BrdU positive expression of hippocampal NSCs in OGD/R group were significantly decreased(P<0.01), while leakage rate of LDH was significantly increased(P<0.01), DAPI staining showed OGD could induce apoptosis. Compared to OGD/R group, NSCs OD value, cell survival rate and Nestin/BrdU positive expression of hippocampal NSCs in senegenin group were significantly increased(P<0.01), and leakage rate of LDH was obviously decreased(P<0.01), DAPI nuclear staining showed that senegenin could inhibit apoptosis. Conclusion: Senegenin can improve the damage of rat hippocampal NSCs injury induced by OGD/R.
Objective: To explore the protective effect of senegenin on the injury of rat hippocampal neural stem cells(NSCs) induced by oxygen-glucose deprivation/reoxygenation(OGD/R) culture in vitro. Methods: Neonatal rat hippocampal NSCs cultured in vitro were randomly divided into control group, OGD/R group, and senegenin group. OGD/R group was cultured by Earle's without glucose in normal culture after hypoxia. Senegenin group was curtured on the basis of OGD/R combined with senegenin. The MTT method was used to detect cell survival rate, the colorimetry was used to detect lactate dehydrogenase(LDH),the DAPI staining method was used to detect apoptosis of NSCs cells, and proliferation of NSCs cells was detected by doublelabeling immunofluorescence method of Nestin/BrdU. Results: Compared with the control group, NSCs OD value, cell survival rate, and Nestin/BrdU positive expression of hippocampal NSCs in OGD/R group were significantly decreased(P<0.01), while leakage rate of LDH was significantly increased(P<0.01), DAPI staining showed OGD could induce apoptosis. Compared to OGD/R group, NSCs OD value, cell survival rate and Nestin/BrdU positive expression of hippocampal NSCs in senegenin group were significantly increased(P<0.01), and leakage rate of LDH was obviously decreased(P<0.01), DAPI nuclear staining showed that senegenin could inhibit apoptosis. Conclusion: Senegenin can improve the damage of rat hippocampal NSCs injury induced by OGD/R.
引文
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[2]吴兴娟,卢晓云,王蕾蕾,等.3-HB协同bFGF诱导促进神经干细胞向神经元方向分化.西安交通大学学报(医学版),2017,38(5):633-638
[3]Chen Y,Huang X,Chen W,et al.Tenuigenin promotes proliferation and differentiation of hippocampal neural stem cells.Neurochem Res,2012,37(4):771-777
[4]师昉,梁志刚,郭紫瑄,等.远志皂苷元促进人神经干细胞体外增生和分化的研究.首都医科大学学报,2013,34(4):559-565
[5]张建平,廖丹琼,储利胜.丹酚酸B对体外模拟脑缺血再灌注损伤新生大鼠海马神经干细胞的保护作用.中华中医药杂志,2017,32(6):2676-2679
[6]李社芳,苗灵娟,李宁,等.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.中国组织工程研究,2017,21(17):2735-2740
[7]马浚宁,高俊玮,侯博儒,等.糖氧剥夺对神经干细胞增殖、分化及凋亡的影响.中国组织工程研究,2015,19(16):876-882
[8]Dong H,Wu S,Hu N,et al.Efficacy of tenuigenin andβ-asarone as augmentations for memantine in the treatment of Alzheimer’s disease.Neuroreport,2018,29(3):203-207
[9]陈玉静,黄小波,陈文强,等.远志皂苷元对新生大鼠海马神经干细胞分化的影响.中国康复理论与实践,2014,20(11):1028-1030
[10]张建平,储利胜,陶水良,等.黄芪甲苷和三七总皂苷配伍对脑缺血再灌注损伤大鼠海马NSCs的保护作用.中成药,2017,39(11):2372-2376