Objective: To investigate the effect of different mechanical loading on the expression of Notch-1, Notch-3, Jagged-1 and Delta-like-1 in growing rabbits' condylar cartilage. Methods: Sixty-four ten-days-old rabbits were randomly divided into experimental and control groups. The rabbits in the experimental groups were fed on a powder diet, while the control groups were fed on a solid diet. The animals were sacrificed after 2, 4, 6 and 8 weeks. Notch-1, Notch-3, Jagged-1 and Delta-like-1 gene and protein expressions were examined by HE, immunohistochemistry, Western blotting and quantitative real-time PCR. Results: At 2, 4, 6, 8 weeks, the anterior part of condylar cartilage in solid diet groups ([318.1±4.3], [342.4±2.6], [364.2±3.2], [380.7±6.0] mm, respectively) were thicker than those in powder diet groups ([275.4±2.6], [301.1±2.0], [322.3±3.3], [366.5±8.4] mm, respectively) (P<0.05). There was no significant difference between the two groups in the middle part (P>0.05). From 2 to 6 weeks, the posterior part of cartilage in solid diet groups ([444.1±1.5], [451.1±0.3], [476.4±5.7] mm, respectively) was thinner than those in power diet groups ([470.4±2.7], [494.3±2.9], [512.3±5.7] mm, respectively) (P<0.05). At 6 weeks, there were more mRNA expressions of Notch-3, Jagged-1 and Delta-like-1 in solid diet group than in power diet group (P<0.05). The Notch-1 protein could be found in all layers of condylar cartilage, especially in the surface of hypertrophic zone. The expressions of Notch-1, Notch-3, and Delta-like-1 protein were increased from 2 to 6 weeks, and decreased at 8 weeks. In solid diet groups, the protein expressions of Notch-1 (at 2, 4, 6, 8 weeks), Notch-3 (at 2, 4, 6 weeks), Jagged-1 (at 2, 4, 6 weeks) and Delta-like-1 (at 4, 6, 8 weeks) were greater than in power diet groups (P<0.05). Conclusions: Low masticatory loading may delay or inhibit the development of condylar cartilage and its growing factors such as Notch-1, Notch-3, Jagged-1 and Delta-like-1. Appropriate masticatory loading plays an important role in normal development of the condyle.
目的:分析咀嚼应力负荷对髁突软骨内Notch信号通路的影响,为进一步研究咀嚼压力对髁突生长发育的影响提供参考。方法:选择64只10日龄幼兔,用随机数字表随机分为硬食组和软食组(每组32只),分别喂颗粒状(硬食组)和粉状(软食组)同种饲料,并于实验2、4、6和8周处死。免疫组化法观察髁突软骨内Notch-1的表达;实时荧光定量PCR(quantitative real-time PCR,qPCR)和蛋白质印迹法检测Notch-1、Notch-3、Jagged-1和Delta-like-1在髁突软骨内的表达。结果:实验2、4、6、8周硬食组髁突前部软骨厚度[分别为(318.1±4.3)、(342.4±2.6)、(364.2±3.2)、(380.7±6.0)mm]均显著大于软食组相应时间点[分别为(275.4±2.6)、(301.1±2.0)、(322.3±3.3)、(366.5±8.4)mm](P<0.05);各时间点两组髁突中部软骨厚度差异均无统计学意义(P>0.05);实验2、4、6周软食组髁突后部软骨厚度[分别为(444.1±1.5)、(451.1±0.3)、(476.4±5.7)mm]均显著大于硬食组[分别为(470.4±2.7)、(494.3±2.9)、(512.3±5.7)mm](P<0.05)。实验6周Notch-3、Jagged-1和Delta-like-1 mRNA表达量硬食组显著大于软食组(P<0.05)。Notch-1蛋白在髁突软骨全层均有表达,肥大细胞层浅层表达最强。实验2~6周Notch-1、Notch-3和Delta-like-1蛋白表达量逐步上升,6周达峰值,8周下降。Notch-1(2、4、6、8周)、Notch-3(2、4、6周)、Jagged-1(2、4、6周)和Delta-like-1(4、6、8周)蛋白表达量硬食组显著大于软食组(P<0.05)。结论:较低的咀嚼负荷可造成髁突软骨生长影响因子Notch-1、Notch-3、Jagged-1和Delta-like-1蛋白分泌减少,Notch信号通路表达迟缓,软骨发育受阻;适当的咀嚼负荷对髁突的正常发育有重要作用。.
Keywords: Bite force; Cartilage; Mandibular condyle; Notch pathway.