摘要
目的探讨红景天苷通过调控APC-Cdh1对永生化神经前体细胞(INPC)增殖分化的影响及机制。方法取对数生长期的INPC用2.5 g/L胰蛋白酶消化传代,处理组加入100μg/mL的红景天苷,对照组无任何处理,培养48 h后,采用流式细胞仪检测细胞周期的分布变化,采用MTT比色法检测INPC细胞增殖情况。加入红景天苷诱导INPC分化,利用免疫荧光检测MAP2的表达及细胞计数观察INPC向神经元分化比例,实时定量RT-PCR检测后期促进复合物(APC)的调控因子(Cdh1)及其下游分化抑制因子2(Id2)mRNA的表达。结果与对照组相比,INPC经过100μg/mL红景天苷处理48 h后,G_(1)~G_(0)期细胞增多(P<0.05),S期减少(P<0.05);MTT显示细胞增殖活力降低(P<0.05);免疫荧光显示MAP2阳性神经元的比例增高(P<0.05);Cdh1 mRNA的表达增加(P<0.05),而Id2 mRNA表达降低(P<0.05)。结论红景天苷100μg/mL能抑制INPC的增殖,促进其向神经元分化,其作用机制可能是通过上调后期促进复合物的调控因子Cdh1、下调分化抑制因子Id2的表达来实现的。
Objective To investigate effect of salidroside mediated APC-Cdh1 on proliferation and differentiation of immortalized neural progenitor cells(INPCs)and its underlying mechanism.Methods The INPCs were treated with salidroside at final concentration of 100μg/mL.After 48h,the cell cycle was examined by fluorescence activated cell sorter.Cell proliferation activity was evaluated using the MTT assay.The fluorescence microscope was used to examine the morphology characteristics and differentiation of INPC according to the expression of MAP2.The expression of Cdh1 and Id2 mRNA was detected by RT-PCR.Results After 48h treatment with salidroside,the cell distribution of G_(O)-G_(1) phase was significantly increased while the cell distribution of S phase was significantly reduced as compared with control group(P<0.05).The proliferation activity of INPCs was significantly reduced(P<0.05).There was dramatic change in the morphology of INPCs,which promoted neurite outgrowth.And the percentage of MAP2 positive cells was significantly upregulated compared with control group(P<0.05).The expression of Cdh1 mRNA was significantly up-regulated,while the expression of Id2 mRNA was significantly down-regulated(P<0.05).Conclusion Salidroside can inhibit the proliferation and promote the differentiation of INPCs,which is mediated by up-regulation of Cdh1 and down-regulation of Id2.
作者
李立
姚文龙
张传汉
LI Li;YAO Wen-long;ZHANG Chuan-han(Department of Physiology,Hubei University of Chinese Medicine,Wuhan 430065,Hubei,China;不详)
出处
《广东医学》
CAS
2022年第4期422-425,共4页
Guangdong Medical Journal
基金
湖北省教育厅科学技术研究项目(Q20162004)。
