口腔医学研究

口腔医学研究 ›› 2021, Vol. 37 ›› Issue (11): 1029-1035.DOI: 10.13701/j.cnki.kqyxyj.2021.11.014

• 颞下颌关节病学研究 • 上一篇    下一篇

电磁脉冲暴露对大鼠下颌髁突软骨细胞程序性坏死的生物学效应研究

马原军1, 何峰1, 王晋2, 刘倩1, 陈晓华1, 何睿1, 李天乐1, 仝昕1, 于世宾1*   

  1. 1.军事口腔医学国家重点实验室,口腔疾病国家临床医学研究中心,陕西省口腔疾病国际联合研究中心,空军军医大学口腔医院解剖生理学教研室 陕西 西安 710032;
    2.空军军医大学军事预防医学系辐射防护医学教研室 陕西 西安 710032
  • 收稿日期:2021-07-19 出版日期:2021-11-28 发布日期:2021-11-22
  • 通讯作者: *于世宾,E-mail:yushibin@fmmu.edu.cn
  • 作者简介:马原军(1989~ ),男,湖北武汉人,硕士在读,主治医师,研究方向:颞下颌关节病的病因及治疗。
  • 基金资助:
    国家自然科学基金面上项目(编号:81970953)陕西省重点研发计划一般项目(编号:2019SF-102)军事口腔医学国家重点实验室自主研究课题(编号:2018ZB02)

Biological Effects of Electromagnetic Pulse on Necroptosis of Mandibular Condylar Chondrocytes in Rats

MA Yuanjun1, HE Feng1, WANG Jin2, LIU Qian1, CHEN Xiaohua1, HE Rui1, LI Tianle1, TONG Xin1, YU Shibin1*   

  1. 1. State Key Laboratory of Military Stomatology, National Clinical Research Center for Oral Diseases, Shaanxi International Joint Research Center for Oral Diseases, Department of Oral Anatomy and Physiology and TMD, School of Stomatology, Air Force Military Medical University, Xi'an 710032, China;
    2. Department of Radiation Protection Medicine, Department of Military Preventive Medicine, Air Force Military Medical University, Xi'an 710032, China
  • Received:2021-07-19 Online:2021-11-28 Published:2021-11-22

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摘要： 目的: 研究不同强度电磁脉冲(electromagnetic pulse,EMP)照射对下颌髁突软骨的生物学效应及软骨细胞程序性坏死(necroptosis)在其中的作用,为今后电磁辐射的应用及治疗提供新思路。方法: SD大鼠随机分为对照组(Sham组)、辐照组(EMP1组和EMP2组),于辐照后3、12、24 h处死取材。苏木精-伊红(HE)、番红O-固绿、Ⅱ型胶原染色评价软骨退变程度;免疫及Western blot检测髁突软骨中Necroptosis关键分子RIPK3、p-MLKL的表达。结果: 与Sham组相比,EMP1组3 h时髁突肥大层大量软骨细胞出现胞核异常深染、增大,12 h、24 h时胞核异常深染现象基本消失;EMP2组细胞形态未见明显改变。与Sham组相比,EMP1组髁突软骨中的Ⅱ型胶原阳性面积百分比显著降低(P< 0.05);但EMP2组Ⅱ型胶原阳性面积百分比未见明显差异(P>0.05)。EMP1组和EMP2组软骨厚度、蛋白多糖阳性面积百分比与Sham组未见明显差异(P>0.05)。与Sham相比,EMP1组髁突软骨中的RIPK3、p-MLKL阳性细胞率及蛋白表达在3 h和24 h显著升高(P<0.01),在12 h与Sham组无明显差异;EMP2组中RIPK3阳性细胞率及蛋白表达显著降低(P<0.01),p-MLKL阳性细胞率及蛋白表达在辐照后3 h显著升高(P<0.05)、12 h无明显差异、24 h显著降低(P<0.05)。结论: 一定剂量的EMP照射可引起髁突软骨的一过性损伤,Necroptosis参与了EMP照射对髁突软骨的瞬时损伤效应。

关键词: 电磁辐射, 电磁脉冲, 髁突, 软骨, 程序性坏死

Abstract: Objective: To explore the biological effects of different high-intensity electromagnetic pulse (EMP) stimulation on mandibular condylar cartilage and the involvement of chondrocyte necroptosis, so as to provide new ideas for the protection and treatment of electromagnetic radiation in the future. Methods: SD rats were randomly divided into Sham group (SHAM group) and irradiation group (EMP1 group and EMP2 group), then were sacrificed at 3 h, 12 h, and 24 h after irradiation. The degree of cartilage degeneration was evaluated by HE staining, Safranin O staining, and type Ⅱ collagen staining. Immunohistochemistry and Western blotting were used to detect the expression of RIPK3 and p-MLKL in condylar cartilage. Results: Compared with SHAM group, a large number of chondrocytes in condylar hypertrophy layer of EMP1 group showed abnormal nuclei hyperchromatism and enlargement at 3 h, and the abnormal hyperchromatism almost disappeared at 12 h and 24 h. There was no significant change in cell morphology in EMP2 group. Compared with SHAM group, the percentage of type Ⅱ collagen positive area in condylar cartilage of EMP1 group was significantly lower (P<0.05). However, there was no significant difference in the percentage of type Ⅱ collagen positive area in EMP2 group (P>0.05). Compared with SHAM group, there was no significant difference in cartilage thickness and percentage of proteoglycan positive area in both EMP1 and EMP2 group (P>0.05). The percentage of positive chondrocytes and protein expression of RIPK3 and p-MLKL in condylar cartilage of EMP1 group were significantly higher than SHAM group at 3 h and 24 h (P<0.01), and with no significant difference from SHAM group at 12 h. In EMP2 group, the percentage of RIPK3 positive chondrocytes and protein expression were significantly decreased (P<0.01), the percentage of p-MLKL positive chondrocytes and protein expression were significantly increased at 3 h (P<0.05), there was no significant difference at 12 h and decreased significantly at 24 h (P<0.05). Conclusion: A certain dose of EMP can cause transient damage of condylar cartilage, in which necroptosis is involved in the transient effect of EMP on condylar cartilage.

Key words: electromagnetic radiation, electromagnetic pulse, condylar, cartilage, necroptosis