摘要
目的探讨地黄多糖诱导大鼠骨髓间充质干细胞（BMSCs）分化为神经样细胞过程中对Noah信号通路的影响。方法以大鼠BMSCs为研究对象，取P3一P5代细胞随机分为正常对照组（CON组）与地黄多糖诱导组（RGP组），诱导后连续培养7d。以免疫细胞化学法和Westernblotting检测诱导后0、1、3、7d各时点Nomhl、Jaggedl蛋白和Notchl胞内段NICD蛋白的表达，Real—timePCR检测Notch信号通路相关分子mRNA。结果免疫荧光法检测显示，RGP组与CON组比较各时点Notchl蛋白表达阳性率差异有统计学意义（P〈0．01）。且随时间变化Notchl蛋白表达阳性率逐渐降低；Jaggedl蛋白阳性细胞率从诱导结束0d到诱导后1d具有明显的上升趋势，1d到7d显著下降，各时点比较差异有统计学意义（P〈0．05），CON组与RGP组各时点比较差异具有统计学意义（P〈0．05）；Westernblotting检测结果与免疫荧光检测结果相似。Real—time PCR检测显示．RGP组Notchl mRNA随时间变化表达下降，Presenilinl表达先降低后略有回升，Hesl表达下降，Mashl表达升高，Jaggedl表达先升高后降低，各时点与CON组比较差异有统计学意义（P〈0．01）。结论地黄多糖在诱导BMSCs向神经样细胞分化过程中抑制Notchl蛋白的表达，并影响Noah信号通路相关基因的表达。
Objective To explore the effect of rehmannia glutinosa olysaccharide （RGP） on rat bone marrow mesen- chymal stem cells （BMSCs） differentiation into neuron-like cells and Notch signaling pathway. Methods Rat BMSCs were experimented as the object, the P3-P5 generations of BMSCs were randomly divided into the control group and RGP group（0.2 mg/mL）, and induced and cultured for 7 days. Immunocytochemistry assay and Western blotting method were used to detect Notchl, Jaggedl protein and intracellular domain of Notch 1 （NICD） expressions at 4 point in time （0,1,3 and 7day） after the induction. The molecular mRNA expression and changes related to the Notch signaling pathway were analyzed by Real-time PCR. Results Immunofluorescence assay showed that the Notchl protein expression of BMSCs decreased over time in the RGP group, positive cell percentage decreased likewise, which was significantly different from the control group（P 〈0.01 ）. The Jaggedl positive cell rate of RGP group increased from induced end 0 d to 1 d after the induction, decreased significantly from ld to 7 d, and there was significant difference between each time point comparison（P 〈 0.05 ）, also different from the control group （P 〈 0.05 ）. Western blotting results were similar to that of the immunofluorescence method. Real-time PCR showed that Notchl mRNA expression decreased over time. Presenilinl decreased and then went up mildly afterwards, Hesl decreased, Mashl increased, and Jaggedl increased then decreased in the RGP group, which were significantly different from the control group at all durations（ P 〈 0.01 ）. Conclusion RGP might restrain Notchl protein expression during BMSCs differentiations into neuron-like cells in vitro, and influence the related molecular mRNA expression of the Notch signaling pathway.
出处
《山东大学学报（医学版）》
CAS
北大核心
2013年第12期1-6,共6页
Journal of Shandong University:Health Sciences
基金
国家自然科学基金(31170930)
辽宁省科技厅自然科学基金(2013022059)
辽宁医学院青年科技启动基金项目(Y2010Z011
Y2011Z010)
关键词
地黄多糖
骨髓间充质干细胞
NOTCH信号通路
诱导
神经样细胞
Rehrnannia glutinosa polysaccharide
Bone marrow mesenchymal stem cells
Notch signal pathway
Induction
Nerve-like cells
