Wu Xiuxiang, Qu Xuebin, Zhang Qiang, Dong Fuxing, Yu Hongli, Yan Chen, Qi Dashi, Wang Meng, Liu Xuan, Yao Ruiqin
Department of Neurobiology, Xuzhou Medical College, Xuzhou, 221002, Jiangsu, People's Republic of China.
Cell Mol Neurobiol. 2014 Apr;34(3):463-71. doi: 10.1007/s10571-014-0030-4. Epub 2014 Feb 12.
The aim of this study was to investigate quercetin's (Qu) ability to promote proliferation and differentiation of oligodendrocyte precursor cells (OPCs) under oxygen/glucose deprivation (OGD)-induced injury in vitro. The results showed that after OGD, OPCs survival rate was significantly increased by Qu as measured by Cell Counting Kit-8. Furthermore, Qu treatment reduced apoptosis of OPCs surveyed by Hoechst 33258 nuclear staining. Qu at 9 and 27 μM promoted the proliferation of OPCs the most by Brdu and Olig2 immunocytochemical staining after OGD 3 days. Also, Qu treatment for 8 days after OGD, the differentiation of OPCs to oligodendrocyte was detected by immunofluorescence staining showing that O4, Olig2, and myelin basic protein (MBP) positive cells were significantly increased compared to control group. Additionally, the protein levels of Olig2 and MBP of OPCs were quantified using western blot and mRNA levels of Olig2 and Inhibitor of DNA binding 2 (Id2) were measured by RT-PCR. Western blot showed a significant increase in Olig2 and MBP expression levels compared with controls after OGD and Qu treatment with a linear does-response curve from 3 to 81 μM. After treatment with Qu compared to its control group, Olig2 mRNA level was significantly up-regulated, whereas Id2 mRNA level was down-regulated. In conclusion, Qu at 3-27 μM can promote the proliferation and differentiation of OPCs after OGD injury and may regulate the activity of Olig2 and Id2.
本研究旨在探讨槲皮素(Qu)在体外氧糖剥夺(OGD)诱导损伤条件下促进少突胶质前体细胞(OPCs)增殖和分化的能力。结果显示,经OGD处理后,用细胞计数试剂盒-8检测发现Qu能显著提高OPCs的存活率。此外,通过Hoechst 33258核染色检测发现Qu处理可减少OPCs的凋亡。在OGD 3天后,采用BrdU和Olig2免疫细胞化学染色法检测发现,9 μM和27 μM的Qu对OPCs增殖的促进作用最为明显。同样,在OGD后用Qu处理8天,通过免疫荧光染色检测OPCs向少突胶质细胞的分化情况,结果显示与对照组相比,O4、Olig2和髓鞘碱性蛋白(MBP)阳性细胞显著增加。此外,采用蛋白质免疫印迹法对OPCs中Olig2和MBP的蛋白水平进行定量分析,并用逆转录聚合酶链反应(RT-PCR)检测Olig2和DNA结合抑制因子2(Id2)的mRNA水平。蛋白质免疫印迹结果显示,与对照组相比,OGD和Qu处理后,Olig2和MBP的表达水平显著升高,且在3至81 μM范围内呈线性剂量反应曲线。与对照组相比,Qu处理后,Olig2的mRNA水平显著上调,而Id2的mRNA水平下调。综上所述,3-27 μM的Qu可促进OGD损伤后OPCs的增殖和分化,并可能调节Olig2和Id2的活性。