Sun Z, Wang L, Peng B
School and Hospital of Stomatology, Wuhan University, Wuhan, China.
Int Endod J. 2014 Dec;47(12):1107-16. doi: 10.1111/iej.12258. Epub 2014 Mar 20.
To investigate the kinetics of GSK3β and p-GSK3β (Ser 9) expression in experimentally induced rat periapical lesions and to explore their possible functions in the pathogenesis of periapical lesions.
Periapical lesions were established in Wistar rats by occlusal pulp exposure in mandibular first molar teeth. The animals were killed on days 0, 7, 14, 21 and 28. Micro-computed tomographic, histological and enzyme histochemical analyses were performed to detect the progression of periapical lesions. Immunohistochemistry, double-dye immunofluorescence and Western blot were performed to determine the expression of GSK3β and p-GSK3β (Ser 9) in periapical tissues.
From day 0 to day 28, the lesion volume and area gradually expanded, and the GSK3β-positive cells gradually ascended. A few p-GSK3β (Ser 9)-positive cells and osteoclasts appeared on day 7 and then climaxed on day 14. The numbers then simultaneously decreased from day 21 to day 28. Western blot analysis revealed that p-GSK3β (Ser 9) and GSK3β proteins were expressed at all time-points. The positive cells and protein expression ratio of p-GSK3β (Ser 9) against GSK3β increased from day 0 to day 14 and then decreased from day 14 to day 28. Finally, double-dye immunofluorescence assay revealed that p-GSK3β (Ser 9)-positive and RANKL-positive cells were co-localized around periapical lesions on days 14 and 28.
GSK3β and p-GSK3β (Ser 9) can be observed and may be involved in alveolar bone resorption and inflammatory response in periapical lesions, as well as associated with periapical lesion pathogenesis.
研究实验性诱导大鼠根尖周病变中糖原合成酶激酶3β(GSK3β)和磷酸化糖原合成酶激酶3β(p - GSK3β,Ser 9)表达的动力学,并探讨它们在根尖周病变发病机制中的可能作用。
通过下颌第一磨牙咬合面牙髓暴露在Wistar大鼠中建立根尖周病变。在第0、7、14、21和28天处死动物。进行微型计算机断层扫描、组织学和酶组织化学分析以检测根尖周病变的进展。采用免疫组织化学、双染免疫荧光和蛋白质印迹法测定根尖周组织中GSK3β和p - GSK3β(Ser 9)的表达。
从第0天到第28天,病变体积和面积逐渐扩大,GSK3β阳性细胞逐渐增多。在第7天出现少量p - GSK3β(Ser 9)阳性细胞和破骨细胞,然后在第14天达到高峰。随后从第21天到第28天数量同时减少。蛋白质印迹分析显示在所有时间点均有p - GSK3β(Ser 9)和GSK3β蛋白表达。p - GSK3β(Ser 9)相对于GSK3β的阳性细胞和蛋白表达比例从第0天到第14天增加,然后从第14天到第28天降低。最后,双染免疫荧光分析显示在第14天和第28天,p - GSK3β(Ser 9)阳性和核因子κB受体活化因子配体(RANKL)阳性细胞在根尖周病变周围共定位。
可观察到GSK3β和p - GSK3β(Ser 9),它们可能参与根尖周病变中的牙槽骨吸收和炎症反应,以及与根尖周病变发病机制相关。