Expression of Angiogenic Factors in Rat Periapical Lesions
Expression of Angiogenic Factors in Rat Periapical Lesions
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DOI：
10.1016/j.joen.2011.11.009
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发表时间：
2012-03-01
影响因子：
4.2
通讯作者：
Okiji, Takashi
中科院分区：
文献类型：
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作者：
Yamanaka, Yusuke;Kaneko, Tomoatsu;Okiji, Takashi
简介：血管生成因子等。因为VEGFR 2（血管内皮细胞生长因子受体2）、BcI-2（促存活和促血管生成信号分子）和chernokine（C-X-C基序）配体1（CXCL 1）（促血管生成趋化因子）可能在促进根尖牙周炎的建立中起关键作用。为了了解这些因素在根尖周炎发病中的作用，我们进行了免疫组织化学和分子生物学分析。方法：Wistar大鼠下颌第一磨牙，采用暴露牙髓的方法建立根尖周炎模型。在14、21和28天后，取出磨牙，包埋成冷冻样品块，并在低温恒温器中切割。正常下颌第一磨牙作为对照。对CD 31（内皮细胞标志物）、BcI-2进行免疫染色，并对VEGFR 2、BcI-2、CXCL 1和CXCR 2信使RNA进行实时聚合酶链反应分析。在实时聚合酶链反应分析中，从用激光捕获显微切割回收的CD 31染色的内皮细胞中提取信使RNA。对于免疫组织化学的统计分析，将免疫染色面积作图，并确定像素计数。然后，计算免疫染色面积在总面积中的百分比。结果：牙髓暴露后21天，CD 31染色区密度增加。另一方面，BcI-2染色区域在第14天（活动性病变扩展期）显示最高密度，然后下降，直到第28天（病变稳定期）。内皮细胞中VEGFR 2、BcI-2、CXCL 1和CXCR 2信使RNA的表达在14天时显示出最高水平，然后下降直至28天时。结论：微血管密度的增加和内皮细胞中VEGFR 2、Bcl-2、CXCL 1和CXCR 2信使RNA表达的上调与实验诱导的根尖周病变的扩展相一致。这些数据表明，这些血管生成因子在病变发展中发挥作用。（J Endod 2012;38：313-317）
Introduction: Angiogenic factors such. as VEGFR2 (vascular endothelial cell growth factor receptor 2), BcI-2 (a prosurvival and proangiogenic signaling molecule), and chernokine (C-X-C motif) ligand 1 (CXCL1) (a proangiogenic chemokine) may have critical roles in enhancing the establishment of apical periodontitis. To understand the role of these factors in the pathogenesis of apical periodontitis, we conducted immunohistochemical and molecular biological analysis. Methods: Apical periodontitis was induced in the lower first molars of Wistar rats by making unsealed pulp exposures. After, 14, 21, and 28 days, the molars were retrieved, embedded as frozen sample blocks, and cut in a cryostat. Normal lower first molars served as controls. lmmunostaining for CD31 (a marker for endothelial cells), BcI-2, and real-time polymerase chain reaction analysis of VEGFR2, BcI-2, CXCL1, and CXCR2 messenger RNA were performed. In the real-time polymerase chain reaction analysis, messenger RNA was extracted from CD31-stained endothelial cells that were retrieved with laser capture microdissection. For statistical analysis of immunohistochemistry, the immunostained area was plotted, and pixel counts were determined. Then, the percentage of the immunostained area in the total area was calculated. Results: The density of the CD31-stained area increased until 21 days after pulp exposure. On the other hand, BcI-2 stained area showed the highest density at 14 days (active lesion expanding phase) and then decreased until 28 days (lesion stability phase). VEGFR2, BcI-2, CXCL1, and CXCR2 messenger RNA expression in endothelial cells showed the highest levels at 14 days and then decreased until 28 days. Conclusions: The increase in microvascular density and the upregulation of VEGFR2, BcI-2, CXCL1, and CXCR2 messenger RNA expression in endothelial cells took place coincidently with the expanding phase of experi- mentally induced periapical lesions. These data suggest that these angiogenic factors play a role in the lesion development. (J Endod 2012;38:313-317)