【摘要】：目的:探討脂多糖調(diào)控Notch信號(hào)通路對(duì)人牙髓干細(xì)胞增殖、分化、凋亡的影響及機(jī)制。方法:從牙髓組織中分離出人牙髓干細(xì)胞(h DPSCs),CCK8實(shí)驗(yàn)檢測(cè)0、0.1、1、10μg/ml的脂多糖處理h DPSCs 1、3、5、7 d后細(xì)胞的增殖情況;RTPCR檢測(cè)1μg/ml的脂多糖處理h DPSCs 0、3、7、14、21 d后礦化相關(guān)基因ALP、DSPP、DMP1的mRNA表達(dá)情況;流式細(xì)胞儀檢測(cè)1μg/ml的脂多糖處理h DPSCs 0、7、14、21 d后的細(xì)胞凋亡情況;Western blot檢測(cè)Cleaved caspase3、Notch1、Hes1的蛋白表達(dá)。結(jié)果:不同濃度的脂多糖刺激h DPSCs 1、3、5 d后細(xì)胞的增殖均無(wú)顯著差異,培養(yǎng)至第7天時(shí),0.1、1、10μg/ml的脂多糖組細(xì)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(shí)ALP、DSPP、DMP1的mRNA表達(dá)與對(duì)照組比較差異均無(wú)統(tǒng)計(jì)學(xué)意義(P0.05),7、14、21 d時(shí)ALP、DSPP、DMP1的mRNA表達(dá)均顯著高于對(duì)照組(P0.01);脂多糖處理h DPSCs 7、14、21 d時(shí)細(xì)胞的凋亡率及Cleaved caspase3、Notch1、Hes1蛋白表達(dá)均顯著高于對(duì)照組(P0.01),21 d時(shí)ALP、DSPP、DMP1的mRNA表達(dá)及細(xì)胞凋亡率和Cleaved caspase3、Notch1、Hes1蛋白表達(dá)均有下降趨勢(shì)。結(jié)論:脂多糖可降低h DPSCs的增殖,促進(jìn)其礦化和凋亡,其機(jī)制與激活Notch信號(hào)通路有關(guān)。
[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(xué)第二醫(yī)院口腔內(nèi)科;
【分類號(hào)】：R781

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