脂多糖对大鼠牙髓细胞ALP、BSP、DSPP表达的研究

发布时间：2018-07-03 09:10

本文选题：牙髓细胞 + 脂多糖　；参考：《青岛大学》2014年硕士论文

【摘要】：目的：本实验通过不同浓度的牙龈卟啉单胞菌(Porphyromonas gingivalis, Pg)脂多糖(lipopolysaccharide, LPS)刺激体外培养的牙髓细胞,检测不同时间下牙髓细胞中各种矿化相关因子(DSPP、ALP、BSP)的表达情况,研究LPS对牙髓细胞矿化能力的影响。方法：采用组织块法获得大鼠牙髓细胞,并用免疫组化方法对培养细胞进行鉴定,以含0.1ng/ml、1ng/ml、10ng/ml、100ng/ml和10000ng/ml牙龈卟啉单胞菌(Porphyromonas gingivalis, Pg) LPS作用牙髓细胞1、3、5d,用荧光定量反转录聚合酶链反应(实时定量PCR)检测DSPP. ALP. BSP mRNA表达的变化,采用spss17.0软件包对实验数据分别进行统计学分析。结果：镜下贴壁后的细胞形态多样,多呈成纤维样细胞形态,还有部分多角形细胞,胞浆突起。实时定量PCR结果显示：与对照组相比,1ng/ml、10ng/ml LPS组,大鼠牙髓细胞DSPP. ALP. BSP的mRNA表达增高,100ng/ml、10000ng/ml LPS组,DSPP、ALP、BSP的mRNA表达均降低；在第1、3、5d时1ng/ml、10ng/ml、100ng/ml和10000ng/ml LPS组：mRNA表达逐渐减少。0.1ng/ml LPS对]mRNA的表达变化无明显差异。3种因子呈现相似的表达变化趋势。结论：低剂量PgLPS有促进牙髓细胞ALP、BSP、DSPP的表达,高剂量时抑制ALP、BSP、DSPP的表达；随着培养时间的延长,促进作用逐渐减弱,抑制作用逐渐增强。
[Abstract]:Objective: to investigate the expression of various mineralization-related factors (DSPP- ALPP-BSP) in dental pulp cells stimulated by different concentrations of Porphyromonas gingivalis (PG) lipopolysaccharide (LPS) in vitro. To study the effect of LPS on the mineralization ability of dental pulp cells. Methods: rat dental pulp cells were obtained by tissue mass method and identified by immunohistochemistry. DSPP was detected by fluorescence quantitative reverse transcriptase polymerase chain reaction (real-time PCR) for 5 days in dental pulp cells treated with 10 ng / ml 10 ng / ml 10000ng/ml and Porphyromonas gingivalis (PG) for 5 days. ALP. The changes of BSP mRNA expression were statistically analyzed by spss17.0 software package. Results: the morphologies of the cells attached to the wall under microscope were various, mostly fibroblast-like cells, and some polygonal cells with cytoplasmic processes. The results of real-time quantitative PCR showed that compared with the control group, DSPPin rat dental pulp cells were treated with 1 ng / ml 10 ng / ml LPS. ALP. The expression of BSP mRNA increased and the expression of BSP mRNA decreased in 100 ng / ml LPS group, but at the first day of 3 days, the mRNA expression of 1 ng / ml 10ng / ml 10000ng/ml decreased gradually. 0.1 ng / ml LPS showed no significant difference in the change of mRNA expression. 3. 3 factors showed a similar trend of expression of BSP mRNA in the LPS-treated group and the LPS group showed no significant difference in the change of the expression of BSP mRNA and the expression of BSP mRNA in the LPS group (P < 0.05), but there was no significant difference in the change of the mRNA expression between the LPS group and the LPS-treated group (P < 0.01 ng / ml). Conclusion: low dose of PgLPS can promote the expression of DSPP in dental pulp cells, inhibit the expression of DSPP in ALP- BSPP at high dose, and weaken the promoting effect and increase the inhibitory effect with the prolongation of culture time.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R781

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