脂多糖调控Notch信号通路作用于人牙髓干细胞增殖、分化、凋亡的实验研究
发布时间：2018-12-25 17:42
【摘要】：目的:探讨脂多糖调控Notch信号通路对人牙髓干细胞增殖、分化、凋亡的影响及机制。方法:从牙髓组织中分离出人牙髓干细胞(h DPSCs),CCK8实验检测0、0.1、1、10μg/ml的脂多糖处理h DPSCs 1、3、5、7 d后细胞的增殖情况;RTPCR检测1μg/ml的脂多糖处理h DPSCs 0、3、7、14、21 d后矿化相关基因ALP、DSPP、DMP1的mRNA表达情况;流式细胞仪检测1μg/ml的脂多糖处理h DPSCs 0、7、14、21 d后的细胞凋亡情况;Western blot检测Cleaved caspase3、Notch1、Hes1的蛋白表达。结果:不同浓度的脂多糖刺激h DPSCs 1、3、5 d后细胞的增殖均无显著差异,培养至第7天时,0.1、1、10μg/ml的脂多糖组细胞的增殖均显著低于0μg/ml的脂多糖组(P0.01);脂多糖处理h DPSCs 3 d时ALP、DSPP、DMP1的mRNA表达与对照组比较差异均无统计学意义(P0.05),7、14、21 d时ALP、DSPP、DMP1的mRNA表达均显著高于对照组(P0.01);脂多糖处理h DPSCs 7、14、21 d时细胞的凋亡率及Cleaved caspase3、Notch1、Hes1蛋白表达均显著高于对照组(P0.01),21 d时ALP、DSPP、DMP1的mRNA表达及细胞凋亡率和Cleaved caspase3、Notch1、Hes1蛋白表达均有下降趋势。结论:脂多糖可降低h DPSCs的增殖,促进其矿化和凋亡,其机制与激活Notch信号通路有关。
[Abstract]:Aim: to investigate the effect and mechanism of lipopolysaccharide (LPS) regulating Notch signaling pathway on proliferation, differentiation and apoptosis of human dental pulp stem cells. Methods: the (h DPSCs), CCK8 assay of human dental pulp stem cells isolated from dental pulp tissue was used to detect the proliferation of human dental pulp stem cells treated with 10 渭 g/ml of 10 渭 g/ml for 7 days after treatment with h DPSCs _ 1 ~ + _ (3). RTPCR was used to detect the mRNA expression of mineralization-related gene ALP,DSPP,DMP1 after 1 渭 g/ml lipopolysaccharide treatment, and flow cytometry was used to detect the apoptosis of 1 渭 g/ml lipopolysaccharide treated h DPSCs 71421 d. The protein expression of Cleaved caspase3,Notch1,Hes1 was detected by Western blot. Results: there was no significant difference in the proliferation of the cells stimulated with different concentrations of lipopolysaccharide for 5 days. At the 7th day of culture, the proliferation of the cells in the 10 渭 g/ml lipopolysaccharide group was significantly lower than that in the 0 渭 g/ml lipopolysaccharide group (P0.01). There was no significant difference in the mRNA expression of ALP,DSPP,DMP1 between the lipopolysaccharide treatment group and the control group on the 3rd day of DPSCs treatment (P0.05). The mRNA expression of ALP,DSPP,DMP1 was significantly higher than that of the control group at 71421 days (P0.01). The apoptotic rate and the expression of Cleaved caspase3,Notch1,Hes1 protein were significantly higher than those of the control group at 21 d after lipopolysaccharide treatment (P0.01), and the mRNA expression, apoptosis rate and Cleaved caspase3,Notch1,Hes1 protein expression of ALP,DSPP,DMP1 decreased at 21 d after lipopolysaccharide treatment. Conclusion: lipopolysaccharide can reduce the proliferation of h DPSCs, promote its mineralization and apoptosis, and its mechanism is related to the activation of Notch signaling pathway.
【作者单位】： 河北医科大学第二医院口腔内科;
【分类号】：R781
[Abstract]:Aim: to investigate the effect and mechanism of lipopolysaccharide (LPS) regulating Notch signaling pathway on proliferation, differentiation and apoptosis of human dental pulp stem cells. Methods: the (h DPSCs), CCK8 assay of human dental pulp stem cells isolated from dental pulp tissue was used to detect the proliferation of human dental pulp stem cells treated with 10 渭 g/ml of 10 渭 g/ml for 7 days after treatment with h DPSCs _ 1 ~ + _ (3). RTPCR was used to detect the mRNA expression of mineralization-related gene ALP,DSPP,DMP1 after 1 渭 g/ml lipopolysaccharide treatment, and flow cytometry was used to detect the apoptosis of 1 渭 g/ml lipopolysaccharide treated h DPSCs 71421 d. The protein expression of Cleaved caspase3,Notch1,Hes1 was detected by Western blot. Results: there was no significant difference in the proliferation of the cells stimulated with different concentrations of lipopolysaccharide for 5 days. At the 7th day of culture, the proliferation of the cells in the 10 渭 g/ml lipopolysaccharide group was significantly lower than that in the 0 渭 g/ml lipopolysaccharide group (P0.01). There was no significant difference in the mRNA expression of ALP,DSPP,DMP1 between the lipopolysaccharide treatment group and the control group on the 3rd day of DPSCs treatment (P0.05). The mRNA expression of ALP,DSPP,DMP1 was significantly higher than that of the control group at 71421 days (P0.01). The apoptotic rate and the expression of Cleaved caspase3,Notch1,Hes1 protein were significantly higher than those of the control group at 21 d after lipopolysaccharide treatment (P0.01), and the mRNA expression, apoptosis rate and Cleaved caspase3,Notch1,Hes1 protein expression of ALP,DSPP,DMP1 decreased at 21 d after lipopolysaccharide treatment. Conclusion: lipopolysaccharide can reduce the proliferation of h DPSCs, promote its mineralization and apoptosis, and its mechanism is related to the activation of Notch signaling pathway.
【作者单位】： 河北医科大学第二医院口腔内科;
【分类号】：R781
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