摘要
用三丁基过氧化氢(t-BHP)构建神经干细胞(NSC)体外衰老模型,探讨人参皂苷Rgl延缓NSC衰老的作用及机制,为寻找延缓NSC衰老新途径提供理论和实验依据。将从新生SD大鼠海马组织中分离纯化的第三代NSC随机分为五组。对照组:在NSC完全培养基中培养2 h;衰老模型组:在对照组基础上加入终浓度为100μmol/L的t-BHP培养2 h;Rg1组:在对照组基础上加入终浓度为10μg/mL的Rg1培养2 h;Rg1抗衰老组:在衰老造模同时加入终浓度为10μg/mL的Rg1培养2h;Rg1治疗衰老组:终浓度为100μmol/L的t-BHP培养2h后再加入终浓度为10μg/mL的Rg1继续培养2 h。MTT法、神经球计数、分化细胞计数以及衰老相关β-半乳糖苷酶(SA-β-Gal)染色阳性神经球计数分析Rg1调控NSC衰老的生物学作用,RT-PCR检测衰老相关基因p16^(1NK4a)、p21^(Cipl/Wafl)mRNA的表达。结果显示,与衰老组比较,Rg1抗衰老组和治疗衰老组NSC的增殖能力和多向分化能力显著增强;衰老特异性SA-β-Gal染色阳性神经球百分比显著降低,p16^(INK4a)、p21^(Cipl/Wafl)mRNA的表达显著下降。提示Rg1具有延缓t-BHP诱导NSC衰老的作用,其机制可能与下调p16^(INK4a)、p21^(Cipl/Wafl)的表达有关。
To provide the theory and experiment foundation for searching the methods of how to delay neural stem cell （NSC） senescence, this study explored the anti-aging effects and the underlying mechanisms of Ginsenoside Rgt on aging model of NSC induced by 100 umol/L tert-butylhydroperoxide （t-BHP）. The third generation of NSCs isolated and purified from neonatal SD rats were divided into 5 groups： control group, aging group （treated with 100 umol/L t-BHP for 2 h）, Rgl group （treated with 10 ug/mL Rgl for 2 h）, Rgl anti-aging group （co-treated with 100 umol/L t-BHP and 10 ug/mL Rgl for 2 h） and Rgl treated-aging group （treated with 10 ug/mL Rgl for 2 h after treated with 100 umol/L t-BHP for 2 h）. The MTT assay, neurospheres counting, differentiated cells counting and senescence-associated β-Galactosidase （SA-β-Gal） staining were used to evaluate the effects of ginsenoside Rgl to regulate NSC senescence. The expressions of senescence associated p16^INK4a and p21^Cipl/Waf1 mRNA were examined by RT-PCR. Compared to aging group, the reproductive activity and the differentiated activity are significantly enhanced; the percentage of SA-β-Gal positive neurospheres and the expressions of p16^INK4a and p21^Cipl/Waf1 mRNA were significantly reduced in Rgl anti-aging group and Rgl treated-aging group. These results suggested that Rgl can regulate the aging process of NSC induced by t-BHE The underlying mechanisms maybe relevant to the down-regulation of expressions of the p16^INK4 and p21^Cipl/Waf1 mRNA.
出处
《中国细胞生物学学报》
CAS
CSCD
2011年第10期1116-1122,共7页
Chinese Journal of Cell Biology
基金
国家自然科学基金(No.30973818)资助项目~~
作者简介
通讯作者。Tel：023—68485968，E-mail：ypwangcq@yahoo．cn
