TNF-α对鼠根尖乳头干细胞增殖及多向分化能力的影响

发布时间：2018-02-21 08:04

  本文关键词： 干细胞 分生组织 肿瘤坏死因子α 根尖乳头干细胞 多向分化 大鼠 Wistar　出处：《重庆医学》2017年14期 　论文类型：期刊论文

【摘要】：目的研究肿瘤坏死因子-α(TNF-α)对鼠根尖乳头干细胞(SCAP)增殖及多向分化能力的影响。方法采用酶消化法结合组织块法获得SCAP,将细胞分为实验组(TNF-α浓度为5、10、20、50ng/mL)和对照组(TNF-α浓度为0ng/mL),使用四甲基偶氮唑蓝(MTT)法检测SCAP的增殖能力;采用茜素红染色及实时定量PCR(qRT-PCR)检测TNF-α对SCAP成骨/成牙本质能力的影响;油红O染色检测TNF-α对SCAP成脂能力的影响;qRT-PCR检测TNF-α对SCAP血管相关基因表达的影响。结果体外培养SCAP符合间充质干细胞来源的特征且具有多向分化能力。MTT结果显示:与对照组相比,各浓度组均能促进SCAP增殖(P0.05),其中10ng/mL TNF-α的促进作用最为明显。茜素红染色结果显示:实验组随着TNF-α浓度的增加,矿化结节逐渐变小,形成数量也逐渐变少。qRT-PCR结果显示:3、7d时,与对照组相比,实验组骨钙蛋白(OC)、牙本质涎磷蛋白(DSPP)、牙本质基质蛋白-1(DMP-1)表达量降低,3d时两组OC、DMP-1比较差异有统计学意义(P0.05);7d时DMP-1比较差异有统计学意义(P0.05);14d时,实验组OC、DMP-1表达量明显降低(P0.05)。油红O染色结果显示:与对照组相比,实验组随着TNF-α浓度的增加,脂滴形成数量逐渐减少。qRT-PCR结果显示:3、7天时,与对照组相比,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表达量明显降低(P0.05)。结论炎性因子TNF-α对SCAP的增殖有明显促进作用但同时不同程度抑制SCAP多向分化能力。
[Abstract]:Objective to study the effect of tumor necrosis factor- 伪 (TNF- 伪) on the proliferation and multidirectional differentiation of rat apical papilla stem cells (SCAPs). Methods SCAPs were obtained by enzyme digestion combined with tissue mass method. The cells were divided into two groups: the concentration of TNF- 伪 in the experimental group was 50 ngmL / L, and the concentration of TNF- 伪 in the control group was 50 ngmL). The proliferative ability of SCAP was measured by tetramethylazolium methylate (MTT) method with a degree of 0 ng / mL. The effect of TNF- 伪 on osteogenesis / dentin formation of SCAP was detected by alizarin red staining and real-time quantitative PCR- qRT-PCR. Effects of TNF- 伪 on the fat-forming ability of SCAP by Oil Red O staining the effects of TNF- 伪 on the expression of Vascular related genes in SCAP were detected by qRT-PCR. Results SCAP cultured in vitro was consistent with the characteristics of mesenchymal stem cells (MSCs) and had the ability to differentiate into different directions. The results showed that compared with the control group, the expression of TNF- 伪 was similar to that of the control group. The results of alizarin red staining showed that with the increase of TNF- 伪 concentration in the experimental group, the mineralized nodules gradually became smaller and the number of the mineralized nodules decreased gradually. The results of qRT-PCR showed that when the concentration of TNF- 伪 increased, the number of mineralized nodules decreased gradually. The results of qRT-PCR showed that 10 ng / mL TNF- 伪 could promote the proliferation of SCAP. Compared with the control group, the expression of osteocalcin, dentin sialophosphorus protein (DSPP) and dentin matrix protein (-1) in the experimental group was significantly lower than that in the control group. There was a significant difference between the two groups in the expression of OCCO-DMP-1 on the 3rd day and the DMP-1 at the 7th day after P0.05D. There was a significant difference between the two groups at P0.0514d. The results of oil red O staining showed that with the increase of TNF- 伪 concentration in the experimental group, the number of lipid droplets gradually decreased. The results of qRT-PCR showed that at the 7th day of 1: 37, compared with the control group, the number of lipid droplets in the experimental group decreased gradually, and that in the control group was higher than that in the control group. The expression of angiopoietin 1, vascular endothelial growth factor A and platelet endothelial cell adhesion molecule-1 decreased significantly (P 0.05). Conclusion the inflammatory factor TNF- 伪 can significantly promote the proliferation of SCAP, but at the same time inhibit the multidirectional differentiation of SCAP.
【作者单位】： 新疆医科大学第二附属医院口腔科;
【基金】：国家自然科学基金资助项目(81460103)
【分类号】：R781.3
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本文编号：1521439