氟对成纤维细胞中结缔组织生长因子表达的影响

发布时间：2018-06-06 17:29

本文选题：氟化物 + 结缔组织生长因子　；参考：《吉林大学》2007年硕士论文

【摘要】： 结缔组织生长因子(connective tissue growth factor,CTGF)可促进软骨细胞和成骨细胞(osteoblast,OB)的增殖和分化,在骨骼发育以及骨量维持方面发挥重要作用。转化生长因子β(transforming growth factor-β,TGF-β)具有与CTGF相似的生物学作用,又是CTGF表达的有效刺激因子。本研究将重点观察氟化物对体外培养成纤维细胞中CTGF mRNA和蛋白表达的特点,尝试分析氟化物作用下TGF-β和CTGF之间的相互关系,从而探讨在氟化物诱导成纤维细胞(fibroblast,FB)成骨功能表达过程中CTGF的作用和作用特点,为解释氟骨症骨周化骨的发生机制提供重要依据。以小鼠成纤维细胞株(L929)和小鼠颅骨成骨样细胞为实验对象,细胞培养方法采用二维和三维两种方法。应用ELISA、免疫组化等方法,检测染氟成纤维细胞CTGF和TGF-β蛋白的表达。结果表明:(1)染氟可刺激二维和三维培养体系中成纤维细胞CTGF蛋白的表达,后者在高氟剂量组明显;(2)PCR检测结果显示,氟化物对两种培养体系中成纤维细胞CTGF mRNA表达均有一定程度的刺激作用,以高氟剂量组(20mg/L)增加幅度大;(3)染氟对二维培养的FB中TGF-β蛋白的表达无明显增强作用,但在染氟早期即明显增强三维培养FB中TGF-β蛋白的表达;(4)相同染氟条件下,二维培养中成骨细胞CTGF mRNA表达与染氟FB的变化趋势相似,但增加程度更为明显。综合分析上述结果,氟化物对FB中CTGF从mRNA到蛋白的表达的刺激作用,提示在氟诱导FB成骨过程中,CTGF可能发挥一定作用,在氟骨症骨周化骨的发生机制的研究中是值得探讨的细胞因子。在氟化物刺激FB中CTGF和TGF-β蛋白的表达方面,未发现明显规律,可能是二者在氟化物的刺激中缺少内在联系,但更可能是由于本实验条件所限,将在进一步的研究中探索改进加氟剂量和时间。氟化物仅对三维培养体系中FB TGF-β蛋白表达具有刺激作用,说明FB在不同的培养条件下,对氟化物的反应可能会有所不同,也提示我们在染氟FB的体外研究中,应用多种培养方法可能得到比较全面的实验结果。
[Abstract]:Connective tissue growth factor (CTGFs) can promote the proliferation and differentiation of chondrocytes and osteoblasts, and play an important role in bone development and bone mass maintenance. Transforming growth factor- 尾 (TGF- 尾) has similar biological effects as CTGF-and is an effective stimulator for CTGF-expression. This study will focus on the characteristics of CTGF mRNA and protein expression in fibroblasts cultured with fluoride, and try to analyze the relationship between TGF- 尾 and CTGF. To explore the role and characteristics of CTGF in the osteogenic expression of fibroblast FB induced by fluoride. In order to provide an important basis for explaining the mechanism of periosseous bone formation in fluorosis, the mouse fibroblast cell line L929) and the osteoblast-like cells of mouse skull were used as experimental objects. The cell culture methods were two-dimensional and three-dimensional. The expression of CTGF and TGF- 尾 protein in fluorinated fibroblasts was detected by ELISAand immunohistochemistry. The results showed that fluoride could stimulate the expression of CTGF protein in fibroblasts in two and three dimensional culture system. Fluoride could stimulate the expression of CTGF mRNA in fibroblasts to a certain extent, and the TGF- 尾 protein expression in FB cultured in high fluoride dose group (20 mg / L) was increased by 20 mg / L) and the expression of TGF- 尾 protein in FB was not significantly enhanced by fluoride. However, the expression of TGF- 尾 protein in FB was significantly enhanced at the early stage of fluoride exposure, and the expression of CTGF mRNA in osteoblasts was similar to that of FB in two dimensional culture, but the increase was more obvious. The stimulating effect of fluoride on the expression of CTGF from mRNA to protein in FB suggests that CTGF may play a certain role in the process of fluorine-induced osteogenesis of FB, and it is a valuable cytokine in the study of the mechanism of periosseous osteogenesis in fluorosis. The expression of CTGF and TGF- 尾 protein in FB stimulated by fluoride did not show obvious regularity, which may be due to the lack of internal relationship between them, but more likely due to the limitation of the experimental conditions. The improvement of fluorine dose and time will be explored in further studies. Fluoride only stimulates the expression of FB TGF- 尾 protein in three dimensional culture system, which indicates that the reaction of FB to fluoride may be different under different culture conditions. More comprehensive experimental results may be obtained by using various cultivation methods.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R363

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