【摘要】：目的:探討脂多糖調控Notch信號通路對人牙髓干細胞增殖、分化、凋亡的影響及機制。方法:從牙髓組織中分離出人牙髓干細胞(h DPSCs),CCK8實驗檢測0、0.1、1、10μg/ml的脂多糖處理h DPSCs 1、3、5、7 d后細胞的增殖情況;RTPCR檢測1μg/ml的脂多糖處理h DPSCs 0、3、7、14、21 d后礦化相關基因ALP、DSPP、DMP1的mRNA表達情況;流式細胞儀檢測1μg/ml的脂多糖處理h DPSCs 0、7、14、21 d后的細胞凋亡情況;Western blot檢測Cleaved caspase3、Notch1、Hes1的蛋白表達。結果:不同濃度的脂多糖刺激h DPSCs 1、3、5 d后細胞的增殖均無顯著差異,培養(yǎng)至第7天時,0.1、1、10μg/ml的脂多糖組細胞的增殖均顯著低于0μg/ml的脂多糖組(P0.01);脂多糖處理h DPSCs 3 d時ALP、DSPP、DMP1的mRNA表達與對照組比較差異均無統(tǒng)計學意義(P0.05),7、14、21 d時ALP、DSPP、DMP1的mRNA表達均顯著高于對照組(P0.01);脂多糖處理h DPSCs 7、14、21 d時細胞的凋亡率及Cleaved caspase3、Notch1、Hes1蛋白表達均顯著高于對照組(P0.01),21 d時ALP、DSPP、DMP1的mRNA表達及細胞凋亡率和Cleaved caspase3、Notch1、Hes1蛋白表達均有下降趨勢。結論:脂多糖可降低h DPSCs的增殖,促進其礦化和凋亡,其機制與激活Notch信號通路有關。
[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.
【作者單位】： 河北醫(yī)科大學第二醫(yī)院口腔內科;
【分類號】：R781

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