TNF-α对鼠根尖乳头干细胞增殖及多向分化能力影响的实验研究

发布时间：2018-04-18 10:54

本文选题：根尖乳头干细胞 + 肿瘤坏死因子-α　；参考：《新疆医科大学》2017年硕士论文

【摘要】：目的:研究肿瘤坏死因子-α对鼠根尖乳头干细胞增殖及多向分化能力的影响,初步探讨肿瘤坏死因子-α对根尖乳头干细胞生物学特性的影响。方法:将细胞分为实验组(肿瘤坏死因子-α浓度为5、10、15、20、30、40、50ng/mL)和对照组(肿瘤坏死因子-α浓度为0ng/mL),使用CCK-8法检测细胞的增殖能力;采用碱性磷酸酶活性、茜素红染色及实时定量PCR检测细胞成骨/成牙本质能力;油红O染色检测细胞成脂能力;实时定量PCR检测血管相关基因的表达。结果:CCK-8显示:各浓度组均能促进根尖乳头干细胞增殖(P0.05),其中10ng/m L肿瘤坏死因子-α的促进作用最为显著;碱性磷酸酶活性显示:各浓度组均能明显降低根尖乳头干细胞的碱性磷酸酶活性(P0.05);茜素红染色显示:随着肿瘤坏死因子-α浓度的增加,矿化结节逐渐变小,形成数量也逐渐变少;实时定量PCR显示:3天,实验组骨钙蛋白、牙本质涎磷蛋白、牙本质基质蛋白-1表达量降低,其中骨钙蛋白、牙本质基质蛋白-1差异具有统计学意义(P0.05),骨涎蛋白表达量增加(P0.05)。7天时,骨钙蛋白、牙本质涎磷蛋白、牙本质基质蛋白-1表达量降低,其中牙本质基质蛋白-1差异具有统计学意义(P0.05),骨涎蛋白表达量与对照组相比仍稍有增加(P0.05);14天时,骨涎蛋白、骨钙蛋白、牙本质基质蛋白-1表达量均明显降低(P0.05),牙本质涎磷蛋白表达量稍有增加(P0.05)。油红O染色结果显示:实验组随着肿瘤坏死因子-α浓度的增加,脂滴形成数量逐渐减少;实时定量PCR显示:3、7天时,血管生成素1、血管内皮生长因子A、血小板内皮细胞黏附分子-1表达量降低(P0.05)。结论:肿瘤坏死因子-α对根尖乳头干细胞的增殖有明显促进作用,且抑制根尖乳头干细胞的成骨/成牙本质、成脂能力及血管相关基因的表达。
[Abstract]:Aim: to study the effects of tumor necrosis factor- 伪 (TNF- 伪) on the proliferation and multidirectional differentiation of rat apical papilla stem cells (RSCs), and to explore the effects of TNF- 伪 on the biological characteristics of root-tip papillary stem cells (RSCs).Methods: the cells were divided into two groups: the experimental group (tumor necrosis factor- 伪 concentration was 5n10151530ng / mL) and the control group (the tumor necrosis factor- 伪 concentration was 0ng / mLL). The proliferative ability of the cells was measured by CCK-8 assay, the activity of alkaline phosphatase was measured by alkaline phosphatase assay, the activity of alkaline phosphatase was measured by alkaline phosphatase assay.Alizarin red staining and real-time quantitative PCR were used to detect osteoblast / dentin ability; oil red O staining was used to detect the adipogenic ability of cells; and real-time quantitative PCR was used to detect the expression of vascular related genes.Results the results showed that every concentration group could promote the proliferation of apical papilla stem cells (P0.05), especially 10ng/m L tumor necrosis factor- 伪 (TNF- 伪).Alkaline phosphatase activity showed that the alkaline phosphatase activity of apical papilla stem cells was significantly decreased in each concentration group, and alizarin red staining showed that the mineralized nodules gradually decreased with the increase of tumor necrosis factor- 伪 concentration.Real time quantitative PCR showed that the expression of osteocalcin, dentin sialophosphorus protein and dentin matrix protein -1 decreased in the experimental group on day 3, in which osteocalcin, osteocalcin, and dentin matrix protein 1 were expressed in the experimental group.The difference of dentin matrix protein 1 was statistically significant (P 0.05). The expression of osteocalcin, dentin sialophosphorus protein and dentin matrix protein 1 decreased after the increase of bone sialoprotein expression.The difference of dentin matrix protein -1 was statistically significant (P 0.05). The expression of bone sialoprotein was still slightly increased after 14 days compared with the control group.The expression of dentine matrix protein-1 was significantly decreased, while that of dentin sialophosphate protein was slightly increased.The results of oil red O staining showed that the number of lipid droplets gradually decreased with the increase of tumor necrosis factor- 伪 concentration in the experimental group, and the real-time quantitative PCR showed that at 7 days after the tumor necrosis factor- 伪 concentration,The expression of angiopoietin 1, vascular endothelial growth factor A and platelet endothelial cell adhesion molecule-1 decreased (P 0.05).Conclusion: tumor necrosis factor- 伪 can significantly promote the proliferation of apical papilla stem cells, and inhibit osteogenesis / dentin formation, fat-forming ability and expression of vascular related genes of apical papilla stem cells.
【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R781

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