不同高强度电磁脉冲对大鼠髁突软骨的生物学效应研究  

A study on the biological effects of electromagnetic pulse with different high-intensity on condylar cartilage in rats

作　　者：马原军 王晋[3] 董青山 何峰 陈晓华 苗辉 于世宾 MA Yuanjun;WANG Jin;DONG Qingshan;HE Feng;CHEN Xiaohua;MIAO Hui;YU Shibin(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;Department of Stomatology,Chinese PLA General Hospi-tal of Central Theater Command,Wuhan 430070,China;Department of Radiation Protection Medicine,Department of Military Preventive Medicine,Air Force Military Medical University,Xi'an 710032,China)

机构地区：[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,第四军医大学口腔医院解剖生理学教研室,陕西西安710032 [2]中国人民解放军中部战区总医院口腔科,湖北武汉430070 [3]第四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

出　　处：《口腔疾病防治》2023年第3期162-170,共9页Journal of Prevention and Treatment for Stomatological Diseases

基　　金：国家自然科学基金面上项目(81970953);陕西省自然科学基础研究计划重点项目(2022JZ-52);陕西省重点研发计划(2021SF-046)。

摘　　要：目的 探索不同高强度电磁脉冲(electromagnetic pulse,EMP)照射对大鼠髁突软骨的生物学效应。方法 SD大鼠随机分为对照组(Sham组)、辐照组(EMP1组:场强500 kV/m、10 Hz;EMP2组:场强270 kV/m、10 Hz),于辐照后1 h、3 h、12 h、24 h、3 d处死取材。通过HE、番红O固绿、Ⅱ型胶原免疫组化和TUNEL染色评估软骨退变程度,通过免疫组化染色及Western blot检测髁突软骨中基质降解因子:基质金属蛋白酶-13(matrix metalloproteinase-13,MMP-13)、血小板结合蛋白基序的解聚蛋白样金属蛋白酶-5(a disintegrin and metalloproteinase with thrombospondin motifs-5,ADAMTS-5)和凋亡关键因子:活化的含半胱氨酸的天冬氨酸蛋白水解酶(cleaved-cysteinyl aspartate specific proteinase,cleaved-Caspase3)的表达。结果 HE染色结果显示,与Sham组相比,EMP1组和EMP2组在辐照后出现了髁突软骨纤维层少量剥脱,EMP1组12、24 h(P<0.01)和EMP2组12、24 h(P<0.05)时软骨中番红O固绿染色阳性面积百分比显著下降,EMP1组3、12 h时Ⅱ型胶原染色阳性面积百分比显著下降(P<0.05,P<0.001),EMP1组1、3、12、24 h和EMP2组1、3、12 h时软骨中TUNEL阳性的凋亡细胞显著增多(P<0.05)。此外,EMP1组、EMP2组不同时间点(3 d组除外)髁突软骨中的MMP-13、ADAMTS-5、cleaved-Caspase3阳性细胞率以及蛋白表达水平均显著升高(P<0.05)。结论 一定程度的高强度EMP能够对髁突软骨造成早期的一过性损伤效应,该效应具有一定的剂量和时间依赖性。Objective To explore the biological effects of electromagnetic pulse(EMP) with different high intensities on condylar cartilage in rats.Methods SD rats were randomly divided into a sham group(Sham) and an irradiation group(EMP1: 500 kV/m, 10 Hz;EMP2: 270 kV/m, 10 Hz). Then, they were sacrificed at 1 h, 3 h, 12 h, 24 h and 3 d after irradiation. The degree of cartilage degeneration was evaluated by HE, safranine O-fast green, type Ⅱ collagen immunohistochemistry and TUNEL staining. Immunohistochemistry and western blot were performed to detect the expression of the matrix degradation factors: matrix metalloproteinase-13(MMP-13), a disintegrin and metalloproteinase with thrombospondin motifs(ADAMTS-5) and the apoptosis key factor cleaved-cysteinyl aspartate specific proteinase(cleaved-Caspase3) in condylar cartilage.Results HE staining showed that, compared with the Sham group, a small amount of exfoliation was found on the fibrous surface layer of the cartilage after irradiation in the EMP1 and EMP2groups. Compared with the Sham group, the percentage of safranine O-fast green-positive area decreased significantly at12 h and 24 h(both P<0.01) in the EMP1 group and 12 h and 24 h in the EMP2 group(both P<0.05);the percentage of type Ⅱ collagen-positive area decreased significantly at 3 h and 12 h(P<0.05, P<0.001) in the EMP1 group. In addition, the number of TUNEL-positive apoptotic cells increased significantly at 1 h, 3 h, 12 h, and 24 h in the EMP1group and 1 h, 3 h, and 12 h in the EMP2 group(P<0.05). Moreover, at different timepoints(except at 3 d) in the EMP1 group and EMP2 group, the percentage of MMP-13, ADAMTS-5-and cleaved Caspase3-positive chondrocytes and their protein levels in condylar cartilage increased significantly after irradiation(P<0.05).Conclusion EMP with a certain degree of high-intensity can induce early transient damage to condylar cartilage. This effect is dose-and timedependent.

关 键 词：电磁辐射 电磁脉冲 颞下颌关节 髁突 软骨 软骨基质 软骨细胞 凋亡 基质金属蛋白酶-13 

分 类 号：R78[医药卫生—口腔医学]