本文關(guān)鍵詞： 干細胞 分生組織 腫瘤壞死因子α 根尖乳頭干細胞 多向分化 大鼠 Wistar　出處：《重慶醫(yī)學(xué)》2017年14期 　論文類型：期刊論文

【摘要】：目的研究腫瘤壞死因子-α(TNF-α)對鼠根尖乳頭干細胞(SCAP)增殖及多向分化能力的影響。方法采用酶消化法結(jié)合組織塊法獲得SCAP,將細胞分為實驗組(TNF-α濃度為5、10、20、50ng/mL)和對照組(TNF-α濃度為0ng/mL),使用四甲基偶氮唑藍(MTT)法檢測SCAP的增殖能力;采用茜素紅染色及實時定量PCR(qRT-PCR)檢測TNF-α對SCAP成骨/成牙本質(zhì)能力的影響;油紅O染色檢測TNF-α對SCAP成脂能力的影響;qRT-PCR檢測TNF-α對SCAP血管相關(guān)基因表達的影響。結(jié)果體外培養(yǎng)SCAP符合間充質(zhì)干細胞來源的特征且具有多向分化能力。MTT結(jié)果顯示:與對照組相比,各濃度組均能促進SCAP增殖(P0.05),其中10ng/mL TNF-α的促進作用最為明顯。茜素紅染色結(jié)果顯示:實驗組隨著TNF-α濃度的增加,礦化結(jié)節(jié)逐漸變小,形成數(shù)量也逐漸變少。qRT-PCR結(jié)果顯示:3、7d時,與對照組相比,實驗組骨鈣蛋白(OC)、牙本質(zhì)涎磷蛋白(DSPP)、牙本質(zhì)基質(zhì)蛋白-1(DMP-1)表達量降低,3d時兩組OC、DMP-1比較差異有統(tǒng)計學(xué)意義(P0.05);7d時DMP-1比較差異有統(tǒng)計學(xué)意義(P0.05);14d時,實驗組OC、DMP-1表達量明顯降低(P0.05)。油紅O染色結(jié)果顯示:與對照組相比,實驗組隨著TNF-α濃度的增加,脂滴形成數(shù)量逐漸減少。qRT-PCR結(jié)果顯示:3、7天時,與對照組相比,血管生成素1、血管內(nèi)皮生長因子A、血小板內(nèi)皮細胞黏附分子-1表達量明顯降低(P0.05)。結(jié)論炎性因子TNF-α對SCAP的增殖有明顯促進作用但同時不同程度抑制SCAP多向分化能力。
[Abstract]:Objective to study the effect of tumor necrosis factor- 偽 (TNF- 偽) 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- 偽 in the experimental group was 50 ngmL / L, and the concentration of TNF- 偽 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- 偽 on osteogenesis / dentin formation of SCAP was detected by alizarin red staining and real-time quantitative PCR- qRT-PCR. Effects of TNF- 偽 on the fat-forming ability of SCAP by Oil Red O staining the effects of TNF- 偽 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- 偽 was similar to that of the control group. The results of alizarin red staining showed that with the increase of TNF- 偽 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- 偽 increased, the number of mineralized nodules decreased gradually. The results of qRT-PCR showed that 10 ng / mL TNF- 偽 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- 偽 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- 偽 can significantly promote the proliferation of SCAP, but at the same time inhibit the multidirectional differentiation of SCAP.
【作者單位】： 新疆醫(yī)科大學(xué)第二附屬醫(yī)院口腔科;
【基金】：國家自然科學(xué)基金資助項目(81460103)
【分類號】：R781.3
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本文編號：1521439