中国实用口腔科杂志 ›› 2023, Vol. 16 ›› Issue (6): 706-712.DOI: 10.19538/j.kq.2023.06.011

• 论著 • 上一篇    下一篇

甲状旁腺激素受体1在大鼠颞下颌关节炎髁突软骨中的表达变化研究

杨    杰1,庄欠志2,于    洋3,王艳霞1   

  1. 1. 潍坊医学院附属医院口腔科,山东 潍坊 261000;2. 潍坊市中医院口腔科,山东  潍坊  261000;3. 潍坊医学院口腔医学院,山东  潍坊  261000
  • 出版日期:2023-11-30 发布日期:2023-12-20
  • 基金资助:
    山东省自然科学基金(ZR2020MH192)

  • Online:2023-11-30 Published:2023-12-20

摘要: 目的    研究甲状旁腺激素受体1(parathyroid hormone receptor 1,PTH1R)在大鼠颞下颌关节骨关节炎(temporomandibular joint osteoarthritis,TMJOA)髁突软骨中的表达变化。方法    研究于2022年7月至2023年1月在潍坊医学院动物实验中心和潍坊市口腔生物医学重点实验室进行。选取8周龄健康雄性SD大鼠48只,将大鼠随机分为实验组和对照组,每组各24只。实验组大鼠用咬合升高法构建TMJOA动物模型,并于造模后的第2、4、6、8周分别处死6只大鼠,解剖分离双侧髁突;对照组大鼠仅进行相同时长的张口动作,其余操作与实验组相同。通过苏木精-伊红和番红O-固绿染色,观察大鼠髁突软骨的组织学变化,按照改良Mankin评分系统对大鼠髁突软骨及软骨下骨拍照并评分;再应用实时定量PCR(quantitative real-time PCR,qRT-PCR)和免疫组化染色,检测大鼠髁突软骨内PTH1R mRNA和蛋白的表达情况。结果    苏木精-伊红和番红O-固绿染色结果显示,所有时间点对照组髁突软骨的组织学形态具有一致性,而实验组颞下颌关节的炎性改变呈现时间依赖性,且Mankin评分随咬合升高时间的延长而增高;qRT-PCR结果显示,第2、4、6、8周,实验组大鼠髁突软骨中PTH1R mRNA的相对表达量均低于相应对照组,差异均有统计学意义(均P < 0.001);免疫组化染色结果显示,随着咬合升高时间的延长,实验组大鼠髁突软骨PTH1R染色强度逐渐减弱;且第2、4、6、8周实验组髁突软骨中PTH1R的积分光密度值均低于相应对照组,差异均有统计学意义(均P < 0.05);实验组Mankin评分与PTH1R mRNA相对表达量及积分光密度值均呈负相关关系(均P < 0.05)。结论    大鼠颞下颌关节炎进展过程中,PTH1R的表达量逐渐下降,与髁突软骨炎症程度呈负相关关系;PTH1R可能是评估炎症进展状态的指标或治疗TMJOA的重要靶点。

关键词: 甲状旁腺激素受体1, 颞下颌关节炎, 髁突软骨, 咬合升高

Abstract: Objective    To study the expressions of parathyroid hormone receptor 1(PTH1R)in condylar cartilage of rats with temporomandibular joint osteoarthritis(TMJOA). Methods    The study was conducted in the Experimental Animal Center of Weifang Medical University and Weifang Key Laboratory of Oral Biomedicine from July 2022 to January 2023. Totally 48 healthy male SD rats aged 8 weeks were selected and divided into experimental group and control group,with 24 rats in each group. The TMJOA animal model was constructed by the elevated occlusion in the experimental group of rats,and 6 rats were euthanized respectively at week 2,4,6 and 8 after modeling,and the bilateral condyles were dissected and isolated. The rats in control group only performed the same duration of open-mouth maneuver,and the rest of the operation was the same as that of the experimental group. Hematoxylin-eosin and safranine O-fast green staining were used to observe the histological changes of condylar cartilage in each group. According to the modified Mankin scoring system,the pathological changes of condylar cartilage and subchondral bone were photographed and scored. Quantitative real-time PCR(qRT-PCR)and immunohistochemistry were used to detect the expression of PTH1R mRNA and protein in the condylar cartilage of each group. Results    Hematoxylin-eosin and safranine O-fast green staining results showed that the histological morphology of the condylar cartilage in the control group was homogeneous at all time points,whereas the inflammatory changes of the condylar cartilage in the experimental group showed a time-dependent pattern and the Mankin score increased with the duration of elevated occlusion. The qRT-PCR results showed that the relative expression of PTH1R mRNA in the condylar cartilage of rats in the experimental group was lower than that in the corresponding control group at week 2,4,6 and 8,and the differences were all statistically significant(all P < 0.001). Immunohistochemical staining results showed that the intensity of PTH1R staining in the condylar cartilage of rats in the experimental groups gradually weakened with the prolongation of elevated occlusion. The value of integral optical density of PTH1R in the condylar cartilage of each experimental group was lower than that of the corresponding control group at week 2,4,6 and 8,and the differences were all statistically significant(all P < 0.05). Mankin scores were negatively correlated with both the relative expressions of PTH1R mRNA and the values of integral optical density in the experimental group(all P < 0.05). Conclusion    During the progression of TMJOA in rats,the expression of PTH1R shows a decreasing trend,which is negatively correlated with the degree of inflammation in the condylar cartilage. PTH1R may be an indicator for evaluating the state of inflammation progression or an important target for the treatment of TMJOA.

Key words: parathyroid hormone receptor 1, temporomandibular joint osteoarthritis, condylar cartilage, elevated occlusion